A recent study by Capaci et al. showed that mutant p53 can interact with HIF1α to induce miR-30d expression which promotes tubulo-vesiculation of Golgi apparatus leading to enhanced vesicular trafficking and secretion ( xref ) ( xref ).

Binding of mutant p53 to ETS2 can promote expression of Pla2g16 or nucleotide synthesis genes required for invasion depending upon the cancer type ( xref ) ( xref , xref ).

Furthermore, the binding of mutant p53 to EGR1 promotes MYO10 expression which drives breast cancer cell invasion ( xref ) ( xref ).

Further, mutant p53 can interact with PELP1 to promote resistance to platinum-based drugs in triple negative breast cancer ( xref ).

A further study reported that mutant p53 enhance the association of mutant p53 and PARP on the replicating DNA ( xref ) ( xref ).

GOF mutant p53 can bind to TopBP1 and attenuate ATR checkpoint response during replication stress ( xref ) ( xref ).

The compound, RETRA disrupts mutant p53-p73 complex restoring p73-dependent transcription and apoptosis ( xref ) ( xref ).

Short Interfering Mutant p53 Peptides (SIMP) can interact with different mutant p53 proteins and release p73, while peptides aptamers (PA) can inhibit mutant p53 transcription ( xref ) ( xref ).

Mutant p53 and p63 complex can increase RAB coupling protein (RCP)-mediated recycling of cell surface growth promoting receptors.

Zhou et al. showed that mutant p53 binds to novel interacting partner AMPKα in glucose starvation conditions and inhibits its activation by other kinases leading to increased aerobic glycolysis, lipid production, and cell growth ( xref ) ( xref ).

This eliminates mitochondria-associated p53 which would otherwise be activated by PINK1 to mediate suppression of Nanog ( xref ) ( xref ).

This eliminates mitochondria associated p53 which would otherwise be activated by PINK1 to mediate suppression of Nanog (XREF_FIG).

Further, the human p53 isoform Delta133p53beta lacking the transactivation domain was observed to promote CSC features in breast cancer cell lines by expression of Sox2, Oct3/4, and Nanog in a Delta133p53beta dependent manner.

Acetylation of p53 at K373 by CBP and p300 leads to dissociation of HDM2 and TRIM24 and subsequent activation of p53 which in turn transcriptionally activates p21, miR-34a, and miR-145 (XREF_FIG).

Further , induction of miR-34a by p53 functionally targets the CSC marker CD44 , thereby inhibiting prostate cancer regeneration and metastasis ( Figure 2 ) ( 74 ) .

Additionally , p53 upregulates miR-34a that represses Notch ( Figure 2 ) and anti-apoptotic Bcl2 thereby promoting differentiation and apoptosis ( 82 ) .

Acetylation of p53 at K373 by CBP/p300 leads to dissociation of HDM2 and TRIM24 and subsequent activation of p53 which in turn transcriptionally activates p21, miR-34a, and miR-145 ( xref ).

Acetylation of p53 at K373 by CBP and p300 leads to dissociation of HDM2 and TRIM24 and subsequent activation of p53 which in turn transcriptionally activates p21, miR-34a, and miR-145 (XREF_FIG).

A recent study by Alam et al. reveals GOF mutant p53 upregulates EFNB2 and activates ephrin B2 reverse signaling to impart enhanced chemoresistance to colorectal cancer cells ( xref ) ( xref ).

Inactivation of p53 disrupts this balance and promotes pluripotency and somatic cell reprogramming .

Inactivation of p53 disrupts this balance and promotes pluripotency and somatic cell reprogramming .

Mutant p53 implicate various context and tissue dependent mechanisms to promote cancer cell invasion and metastasis.

Various transcription factors such as NF-Y, SREBPs, ETS, and EGFR1 play crucial role in mutant p53 driven invasion and metastasis.

Mutant p53 implicate various context and tissue dependent mechanisms to promote cancer cell invasion and metastasis.

Various transcription factors such as NF-Y, SREBPs, ETS, and EGFR1 play crucial role in mutant p53 driven invasion and metastasis.

This suggests that acetylation at K320 and K373 can alter the structure of mutant p53 and restore wild type p53 functions.

Mutant p53 can also induce YAP and TAZ nuclear localization by interacting with SREBP and activating the mevalonate pathway.

Mutant p53 can also promote proliferation by inducing the REG-gamma proteosome pathway in association with p300 (XREF_FIG).

While wild type p53 suppresses inflammatory response by inhibiting the production of cytokines and antagonizing NF-kB activity, mutant p53 on the other hand enhances NF-kB activity in response to TNF-alpha and promotes inflammation (XREF_FIG).

GOF mutant p53 can modify the tumor microenvironment and has been found to support chronic inflammation.

Several evidence demonstrate that mutant p53 promotes glycolysis and reprograms the cellular metabolism of cancer cells.

In absence of AMPK, mitochondrial stress augments aerobic glycolysis, also called " Warburg effect " in tumor cells, which is promoted by mutant p53.

However, whether mutant p53 induced EMT trigger stemness properties in cancer cells, is still quite unexplored.

Although these studies highlight that mutant p53 mediated EMT phenotype confer stemness in cancer cells, however, there is still a lot to explore in context of molecular mechanisms of mutant p53 driven stemness through activation of EMT genes.

Gain-of function mutant p53 further promotes EMT and stemness phenotypes by activating genes regulating them.

The sustained activation of NF-kB signaling by mutant p53 not only elevate inflammatory response but also protects the cancer cells from cytotoxic effects of tumor microenvironment by activating pro survival pathways.

While wild type p53 suppresses inflammatory response by inhibiting the production of cytokines and antagonizing NF-kB activity, mutant p53 on the other hand enhances NF-kB activity in response to TNF-alpha and promotes inflammation (XREF_FIG).

Mutant p53 can also induce YAP and TAZ nuclear localization by interacting with SREBP and activating the mevalonate pathway.

Interaction of mutant p53 to SREBPs activates mevalonate pathway that promotes invasion in breast cancer cells (XREF_FIG).

Similarly, p53 activation by nutlin leads to transcriptional activation of p21 that cause cell cycle arrest and induces differentiation in human ESCs.

Similarly , p53 activation by nutlin leads to transcriptional activation of p21 that cause cell cycle arrest and induces differentiation in human ESCs ( 35 ) .

Acetylation of p53 at K373 by CBP/p300 leads to dissociation of HDM2 and TRIM24 and subsequent activation of p53 which in turn transcriptionally activates p21, miR-34a, and miR-145 ( xref ).

The enhancer of zeste homolog 2 (EZH2) is a catalytic subunit of the polycomb repressive complex 2 (PRC2), acts as a histone methyltransferase and induces the trimethylation of histone H3 lysine 27 (H3K27me3) in the promoter of many key genes.

The enhancer of zeste homolog 2 (EZH2) is a histone methyltransferase and induces the trimethylation of histone H3 lysine 27 (H3K27me3) in the promoter of many key genes; EZH2 acts as a transcriptional repressor and is an epigenetic regulator for several cancers.

The enhancer of zeste homolog 2 (EZH2) is a histone methyltransferase and induces the trimethylation of histone H3 lysine 27 (H3K27me3) in the promoter of many key genes; EZH2 acts as a transcriptional repressor and is an epigenetic regulator for several cancers.

The enhancer of zeste homolog 2 (EZH2) is a catalytic subunit of the polycomb repressive complex 2 (PRC2), acts as a histone methyltransferase and induces the trimethylation of histone H3 lysine 27 (H3K27me3) in the promoter of many key genes.

The enhancer of zeste homolog 2 (EZH2) is a histone methyltransferase and induces the trimethylation of histone H3 lysine 27 (H3K27me3) in the promoter of many key genes; EZH2 acts as a transcriptional repressor and is an epigenetic regulator for several cancers.

The inhibition of EZH2 aggravated cisplatin induced injury in renal tubular cells by inactivating mTOR complexes.

Notably, the effect of EZH2 inhibition-induced inactivation of mTOR complexes was not completely reversed after Deptor depletion, suggesting that Deptor might not be the only target of EZH2 epigenetic regulation.

After inhibiting Deptor expression with siRNA, the effect of EZH2 inhibition-induced inactivation of mTOR complexes was reversed.

The present study showed that the inhibition of EZH2 induced apparent apoptosis in cultured NRK-52E cells, as demonstrated by flow cytometry and the concomitant increase of a pro apoptosis protein (cleaved-caspase 3) and decrease of anti-apoptosis proteins (Bcl-2 and HuR).

These data suggested that EZH2 inhibition induced notable apoptosis in NRK-52E cells.

Inhibition of EZH2 induced apoptosis in NRK-52E cells.

Thus, the regulation of Deptor expression by EZH2 may control cell growth and proliferation through mTOR complex pathways.

These data indicated that EZH2 inhibition decreased mTORC1 and mTORC2 activity by up-regulating Deptor expression.

In the present study, we identified that the inhibition of EZH2 with 3-deazaneplanocin A (DZNep) upregulated the transcription of Deptor by decreasing the H3K27me3 methylation level in its promoter region and reduced the activity of mTORC1 and mTORC2, resulting in apoptosis of NRK-52E cells.

These data suggested that EZH2 inhibition increased the transcription of Deptor by modifying H3K27me3 in its promoter region, subsequently inhibited mTORC1 and mTORC2 activities, downregulated the expression of apoptosis suppressor genes, and finally led to apoptosis in renal tubular cells.

In summary, our results showed that EZH2 inhibition increased the transcription level of Deptor by decreasing the level of trimethylation of H3K27 in the Deptor promoter region, subsequently inhibited the activities of mTORC1 and mTORC2, downregulated the expression of HuR and Bcl-2, and finally led to apoptosis in renal tubular cells.

EZH2 bound the Deptor promoter region and then regulated its transcriptional level.

A ChIP assay demonstrated that EZH2 bound the promoter region of Deptor, an endogenous inhibitor of mTORC1 and mTORC2, and regulated the transcription of Deptor by modulating H3K27me3 in its promoter region.

The binding of EZH2 to the Deptor promoter was determined by ChIPassay.

EZH2 bound the Deptor promoter region and then regulated its transcriptional level (Fig.  xref d).

A ChIP assay demonstrated that EZH2 bound the promoter region of Deptor, an endogenous inhibitor of mTORC1 and mTORC2, and regulated the transcription of Deptor by modulating H3K27me3 in its promoter region.

The binding of EZH2 to the Deptor promoter was determined by ChIP assay.

These data indicated that EZH2 inhibition decreased the activity of mTORC1 and mTORC2 activity.

Inhibition of EZH2 decreased the activity of mTORC1 and mTORC2.

These data indicated that HuR is situated between the mTOR complexes and Bcl-2 and that EZH2 inhibition might inactivate mTORC1 and mTORC2 in some manner, thus downregulating HuR and Bcl-2 expression and leading to cell apoptosis.

These data indicated that EZH2 inhibition decreased the activity of mTORC1 and mTORC2 activity.

Inhibition of EZH2 decreased the activity of mTORC1 and mTORC2.

These data indicated that HuR is situated between the mTOR complexes and Bcl-2 and that EZH2 inhibition might inactivate mTORC1 and mTORC2 in some manner, thus downregulating HuR and Bcl-2 expression and leading to cell apoptosis.

Interestingly, another report indicated that CDK1 phosphorylates EZH2 at Thr 487, thereby leading to suppression of H3K27 trimethylation and consequent derepression of target gene expression xref .

Many previous studies have shown that the canonical function of EZH2 is mediating trimethylation of histone H3 lysine 27 and inhibiting downstream target genes XREF_BIBR, XREF_BIBR, XREF_BIBR.

Many previous studies have shown that the canonical function of EZH2 is mediating trimethylation of histone H3 lysine 27 and inhibiting downstream target genes XREF_BIBR, XREF_BIBR, XREF_BIBR.

Additionally, EZH2 specific microRNA-98 could effectively inhibit cell proliferation in vitro and regulate the pRb-E2F pathway in human epithelial ovarian cancer stem cells (EOCSCs) XREF_BIBR.

The results showed that EZH2 inhibition significantly increased cisplatin induced cell apoptosis and necrosis and reduced the number of colonies formed by SK-3rd cells compared with control groups.

In addition, ChIP assays showed that EZH2 bound directly to region 1 and region 2 within the CHK1 promoter region in both SKOV3 and A2780 cells, indicating that EZH2 upregulated CHK1 expression by targeting the CHK1 promoter.

However, EZH2 DeltaSET mutant increased CHK1 level as efficiently as wild-type EZH2 (XREF_SUPPLEMENTARY G), suggesting that the histone methyltransferase activity may not be required for CHK1 activation.

EZH2 transcriptionally upregulates CHK1 expression by directly binding to the CHK1 promoter, and inhibition of CHK1abrogates G2/M checkpoints and promotes DNA damaging agent induced cell death in EOCSC.

EZH2 knockdown reduces CSCs and inhibits chemoresistance and tumorigenesis in ovarian cancer cells To examine the effect of EZH2 on CSC populations in ovarian cancer , we first compared the protein level of EZH2 in SKOV3 , SK-1st , SK-2nd and SK-3rd cells by Western blot and found a gradual increase in the EZH2 level ( Figure 2A ) .

EZH2 knockdown reduces CSCs and inhibits chemoresistance and tumorigenesis in ovarian cancer cells To examine the effect of EZH2 on CSC populations in ovarian cancer , we first compared the protein level of EZH2 in SKOV3 , SK-1st , SK-2nd and SK-3rd cells by Western blot and found a gradual increase in the EZH2 level ( Figure 2A ) .

Similarly, a previous study revealed that EZH2 promotes breast CSC expansion and leads to tumor initiation XREF_BIBR.

EZH2 promotes CSC properties and chemoresistance by upregulating CHK1.

In addition , it was reported that EZH2 knockdown significantly reduces the frequency of CSCs in pancreatic ductal adenocarcinoma 38 .

Conclusions : Our data revealed a previously unidentified functional and mechanistic link between EZH2 levels , CHK1 signaling activation , and ovarian CSCs and provided strong evidence that EZH2 promotes ovarian cancer chemoresistance and recurrence .

Conclusions : Our data revealed a previously unidentified functional and mechanistic link between EZH2 levels , CHK1 signaling activation , and ovarian CSCs and provided strong evidence that EZH2 promotes ovarian cancer chemoresistance and recurrence .

In our study , we uncovered a mechanism by which EZH2 directly occupies the promoter region of CHK1 and induces its activation in epithelial ovarian cancer , which is consistent with a recent study showing that EZH2 functioned in activating NOTCH1 signaling by directly binding to the NOTCH1 promoter in breast cancer 37 .

EZH2 activates CHK1 signaling to promote ovarian cancer chemoresistance by maintaining the properties of cancer stem cells Background : Ovarian cancer is a fatal malignant gynecological tumor .

A luciferase reporter assay and chromatin immunoprecipitation assay were performed to identify activation of CHK1 by EZH2 .

By providing previously unidentified evidence that EZH2 promotes the platinum resistance of ovarian CSCs by directly activating CHK1 signaling, our work paves the way for targeting EZH2 to reverse recurrence and platinum resistance in ovarian cancer.

EZH2 activates CHK1 signaling to promote ovarian cancer chemoresistance by maintaining the properties of cancer stem cells.

EZH2 promotes CSC properties and chemoresistance by upregulating CHK1.

A luciferase reporter assay and chromatin immunoprecipitation assay were performed to identify activation of CHK1 by EZH2.

A luciferase reporter assay and chromatin immunoprecipitation assay were performed to identify activation of CHK1 by EZH2.

A luciferase reporter assay and chromatin immunoprecipitation assay were performed to identify activation of CHK1 by EZH2 .

EZH2 transcriptionally upregulates CHK1 expression by directly binding to the CHK1 promoter , and inhibition of CHK1abrogates G2 / M checkpoints and promotes DNA damaging agent-induced cell death in EOCSC .

In addition , ChIP assays showed that EZH2 bound directly to region 1 and region 2 within the CHK1 promoter region in both SKOV3 and A2780 cells ( Figure 3F-H ) , indicating that EZH2 upregulated CHK1 expression by targeting the CHK1 promoter .

Upregulation of CHK1 partially relieved these effects on the cell cycle and apoptosis caused by EZH2 depletion.

Upregulation of CHK1 partially relieved these effects on the cell cycle and apoptosis caused by EZH2 depletion.

The protein expression studies have further revealed that vanillin reduces the CDK6 expression and induces apoptosis in the cancer cells .

Enzyme inhibition and fluorescence-binding studies showed that vanillin inhibits CDK6 with an half maximal inhibitory concentration = 4.99 muM and a binding constant ( K ) 4.1 x 10 7 M -1 .

Instructively, premature senescence in NP-MSCs could be largely alleviated using ASIC inhibitors, suggesting both ASIC1 and ASIC3 act decisively upstream to activate senescence programming pathways including p53-p21/p27 and p16-Rb1 signaling.

These events corresponded to the RB1-SIAH3 fusion and a novel RB1 rearrangement expected to correlate with the complete absence of RB1 protein expression.

RNA-sequencing revealed multiple fusions clustered within 13q14.1-q21.3, including a novel in-frame fusion of RB1-SIAH3 predicted to prematurely truncate the RB1 protein.

However, blocking CD32 but not CD64 to inhibit CRP induced FLS proliferation, invasiveness, and proinflammatory cytokine CXCL8 production revealed a major role for CD32 signaling in synovial inflammation, although CRP via CD64, not CD32, to induce MMP9 expression was noticed.

Interestingly, CRP-induced MMP9 expression and invasion on RA-FLSs were p38-dependent as addition of a p38 inhibitor (SB202190) but not a NF-κB inhibitor (PDTC) was capable of inhibiting CRP-induced MMP9 expression and cell invasion.

In contrast, blockade of CD16 produced no inhibitory effect on CRP-induced expression of CXCL8, CCL2, MMP9 and IL-6 by RA-FLSs ( xref ), suggesting that CRP may not signal through the CD16 to induce joint inflammation in vitro .

In contrast, blockade of CD16 produced no inhibitory effect on CRP-induced expression of CXCL8, CCL2, MMP9 and IL-6 by RA-FLSs ( xref ), suggesting that CRP may not signal through the CD16 to induce joint inflammation in vitro .

However, CRP- induced expression of CXCL8 was CD32-dependent as it was blunted by the antibody against CD32, whereas CRP-induced MMP9 was blocked by the antibody to CD64, demonstrating that differential signaling mechanisms for CRP in regulating CXCL8 and MMP9 expression in RA-FLSs.

As shown in XREF_FIG, CRP induced expression of CCL2 and IL-6 was blocked by either neutralizing antibody to CD32 or CD64 or both, suggesting that CRP signals through both CD32 and CD64 to induce expression of CCL2 and IL-6.

In contrast, blockade of CD16 produced no inhibitory effect on CRP-induced expression of CXCL8, CCL2, MMP9 and IL-6 by RA-FLSs ( xref ), suggesting that CRP may not signal through the CD16 to induce joint inflammation in vitro .

As shown in xref , CRP-induced expression of CCL2 and IL-6 was blocked by either neutralizing antibody to CD32 or CD64 or both, suggesting that CRP signals through both CD32/CD64 to induce expression of CCL2 and IL-6.

As shown in XREF_FIG, CRP induced expression of CCL2 and IL-6 was blocked by either neutralizing antibody to CD32 or CD64 or both, suggesting that CRP signals through both CD32 and CD64 to induce expression of CCL2 and IL-6.

In contrast, blockade of CD16 produced no inhibitory effect on CRP-induced expression of CXCL8, CCL2, MMP9 and IL-6 by RA-FLSs ( xref ), suggesting that CRP may not signal through the CD16 to induce joint inflammation in vitro .

As shown in xref , CRP-induced expression of CCL2 and IL-6 was blocked by either neutralizing antibody to CD32 or CD64 or both, suggesting that CRP signals through both CD32/CD64 to induce expression of CCL2 and IL-6.

It has been reported that CRP-induced cytokine expression is also regulated by TWIST transcriptionally in myeloma cells ( xref ).

We thus blocked CD32 or CD64 or both with neutralizing antibodies to differentially determine the signaling mechanisms of CRP-induced cytokine expression.

Thus, it is highly possible that high concentration of CRP in synovial fluid in patients with RA may directly bind primarily CD32 to activate p38 MAP kinase and NF-kappaB signaling in RA-FLSs to differentially regulate synovial inflammation.

In vitro studies confirmed this notion and found that CRP was able to upregulate both CD32 and CD64 and induced FLS proliferation, invasion, and pro inflammatory expression by increasing production of CCL2, CXCL8, IL-6, MMP2, MMP9 while suppressing an anti-inflammatory cytokine IL-10 expression.

RA-FLS was a major cell type responsible for CRP production in RA patients, accounting for more than 65% of CRP producing cells as identified by co-expressing CRP and vimentin in the inflamed synovial tissues in patients with RA.

In vitro studies confirmed this notion and found that CRP was able to upregulate both CD32 and CD64 and induced FLS proliferation, invasion, and pro inflammatory expression by increasing production of CCL2, CXCL8, IL-6, MMP2, MMP9 while suppressing an anti-inflammatory cytokine IL-10 expression.

As shown in XREF_FIG, multiplex cytokine assay kits assays showed that addition of CRP dose-dependently upregulated CCL2, CXCL8, IL-6, MMP2, MMP9 in RA-FLS but not in HFLS, although expression of IL-1beta and TNFalpha was not significantly changed (XREF_FIG).

In vitro studies confirmed this notion and found that CRP was able to upregulate both CD32 and CD64 and induced FLS proliferation, invasion, and pro inflammatory expression by increasing production of CCL2, CXCL8, IL-6, MMP2, MMP9 while suppressing an anti-inflammatory cytokine IL-10 expression.

As shown in XREF_FIG, multiplex cytokine assay kits assays showed that addition of CRP dose-dependently upregulated CCL2, CXCL8, IL-6, MMP2, MMP9 in RA-FLS but not in HFLS, although expression of IL-1beta and TNFalpha was not significantly changed (XREF_FIG).

In vitro studies confirmed this notion and found that CRP was able to upregulate both CD32 and CD64 and induced FLS proliferation, invasion, and pro inflammatory expression by increasing production of CCL2, CXCL8, IL-6, MMP2, MMP9 while suppressing an anti-inflammatory cytokine IL-10 expression.

As shown in XREF_FIG, multiplex cytokine assay kits assays showed that addition of CRP dose-dependently upregulated CCL2, CXCL8, IL-6, MMP2, MMP9 in RA-FLS but not in HFLS, although expression of IL-1beta and TNFalpha was not significantly changed (XREF_FIG).

In vitro studies confirmed this notion and found that CRP was able to upregulate both CD32 and CD64 and induced FLS proliferation, invasion, and pro inflammatory expression by increasing production of CCL2, CXCL8, IL-6, MMP2, MMP9 while suppressing an anti-inflammatory cytokine IL-10 expression.

As shown in XREF_FIG, multiplex cytokine assay kits assays showed that addition of CRP dose-dependently upregulated CCL2, CXCL8, IL-6, MMP2, MMP9 in RA-FLS but not in HFLS, although expression of IL-1beta and TNFalpha was not significantly changed (XREF_FIG).

In vitro studies confirmed this notion and found that CRP was able to upregulate both CD32 and CD64 and induced FLS proliferation, invasion, and pro inflammatory expression by increasing production of CCL2, CXCL8, IL-6, MMP2, MMP9 while suppressing an anti-inflammatory cytokine IL-10 expression.

As shown in XREF_FIG, multiplex cytokine assay kits assays showed that addition of CRP dose-dependently upregulated CCL2, CXCL8, IL-6, MMP2, MMP9 in RA-FLS but not in HFLS, although expression of IL-1beta and TNFalpha was not significantly changed (XREF_FIG).

In vitro studies confirmed this notion and found that CRP was able to upregulate both CD32 and CD64 and induced FLS proliferation, invasion, and pro inflammatory expression by increasing production of CCL2, CXCL8, IL-6, MMP2, MMP9 while suppressing an anti-inflammatory cytokine IL-10 expression.

This was further confirmed by the ability of pre-treating RA-FLS with a NF-kappaB inhibitor, PDTC (100 mumol/L) to inhibit CRP induced proliferation (XREF_FIG) and upregulation of CXCL8, CCL2.

Here we tested the hypothesis that CRP may be produced locally by FLSs and functions to induce the synovial inflammation in patients with RA.

CRP may promote synovial inflammation via mechanism associated with activation of CD32/64- p38 and NF-kappaB signaling.

In the present study, we found that CRP signaled primarily through CD32, to a less extent of CD64, to differentially regulate joint inflammation.

CRP Promotes RA-FLS Pro inflammatory Response Differentially via the CD32/64-p 38 and NF-kappaB-Dependent Mechanisms in vitro.

CRP can induce synovial inflammation via mechanisms associated with activation of CD32/64-p 38 and NF-kappaB signaling.

This was supported by the findings that CRP induced activation of p38 MAP kinase and NF-kappaB signaling was blunted by neutralizing antibodies against CD32 but not CD64.

To examine whether CRP induces NF-kappaB nuclear translation, immunofluorescence and subcellular fractionation were performed.

Resveratrol significantly suppresses the secretion of TNF-alpha and nitric oxide in LPS stimulated rat cortical microglia and N9 microglial cells (Bi et al., 2005) and also inhibits the production of TNF-alpha, IL-1, IL-6, IL-12 and IFN-gamma by splenic lymphocytes and macrophages (Gao et al., 2001; Kowalski et al., 2005).

Resveratrol significantly suppresses the secretion of TNF-alpha and nitric oxide in LPS stimulated rat cortical microglia and N9 microglial cells (Bi et al., 2005) and also inhibits the production of TNF-alpha, IL-1, IL-6, IL-12 and IFN-gamma by splenic lymphocytes and macrophages (Gao et al., 2001; Kowalski et al., 2005).

Resveratrol significantly suppresses the secretion of TNF-alpha and nitric oxide in LPS stimulated rat cortical microglia and N9 microglial cells (Bi et al., 2005) and also inhibits the production of TNF-alpha, IL-1, IL-6, IL-12 and IFN-gamma by splenic lymphocytes and macrophages (Gao et al., 2001; Kowalski et al., 2005).

Kaempferol and quercetin reduce the activity of iNOS and COX-2 by suppressing the signalling of STAT-1, NF-kappa B and AP-1 in LPS- or cytokine stimulated macrophages and HUVECs (Crespo et al., 2008; Hamalainen et al., 2007).

Kaempferol and quercetin reduce the activity of iNOS and COX-2 by suppressing the signalling of STAT-1, NF-kappa B and AP-1 in LPS- or cytokine stimulated macrophages and HUVECs (Crespo et al., 2008; Hamalainen et al., 2007).

Kaempferol, quercetin, and luteolin also afford protection against oxidative stress by inducing the expression of HO-1, however, apigenin inhibits HO-1 induction.

Our in vitro experiments show that quercetin inhibits the intrinsic apoptotic pathway by increasing the ratio of BCL-2 and BAX, thus preventing apoptosis and DNA damage in RPE cells under oxidative stress.

Quercetin could not effectively downregulate the increased transcripts of the ocular inflammatory mediators Tnf-alpha, Cox-2 and Inos.

Our in vitro experiments show that quercetin inhibits the intrinsic apoptotic pathway by increasing the ratio of BCL-2 and BAX, thus preventing apoptosis and DNA damage in RPE cells under oxidative stress.

Quercetin could not effectively downregulate the increased transcripts of the ocular inflammatory mediators Tnf-alpha, Cox-2 and Inos.

Additionally, quercetin could not effectively suppress ocular transcripts of the pro apoptotic factors Fas, FasL and Caspase-3.

Quercetin could not effectively downregulate the increased transcripts of the ocular inflammatory mediators Tnf-alpha, Cox-2 and Inos.

Additional studies showed that EGCG, luteolin and structural analogs of luteolin such as quercetin, chrysin, and eriodictyol, also inhibited TRIF signaling pathway by targeting TBK1 kinase [XREF_BIBR, XREF_BIBR].

In contrast, resveratrol, EGCG, luteolin, and structural analogs of luteolin specifically inhibit TLR3 and TLR4 signaling by targeting TANK binding kinase 1 (TBK1) and receptor interacting protein 1 (RIP1) in Toll/IL -1 receptor domain containing adaptor inducing IFN-beta (TRIF) complex.

In contrast, resveratrol, EGCG, luteolin and structural analogs of luteolin, specifically inhibit TLR3 and TLR4 signaling by targeting TBK1 and RIP1 in TRIF complex.

In contrast, resveratrol, EGCG, luteolin, and structural analogs of luteolin specifically inhibit TLR3 and TLR4 signaling by targeting TANK binding kinase 1 (TBK1) and receptor interacting protein 1 (RIP1) in Toll/IL -1 receptor domain containing adaptor inducing IFN-beta (TRIF) complex.

In contrast, resveratrol, EGCG, luteolin and structural analogs of luteolin, specifically inhibit TLR3 and TLR4 signaling by targeting TBK1 and RIP1 in TRIF complex.

Curcumin, helenalin, and cinnamaldehyde with alpha, beta unsaturated carbonyl groups, or sulforaphane with an isothiocyanate group, inhibit TLR4 activation by interfering with cysteine residue mediated receptor dimerization, while resveratrol, with no unsaturated carbonyl group, did not.

Curcumin, helenalin, cinnamaldehyde and sulforaphane, containing alpha, beta unsaturated carbonyl or isothiocyanate group, respectively, that are known to interact with free SH groups in cysteine residues, but not resveratrol (with no unsaturated carbonyl group), inhibit TLR4 activation by interfering with TLR4 receptor dimerization.

However, curcumin did not inhibit interferon regulatory factor 3 (IRF3) activation induced by another immediate TLR4 downstream component TIR-domain-containing adaptor inducing interferon-beta (TRIF), suggesting that the target of curcumin is the receptor itself, but not the downstream components of TRIF pathway [XREF_BIBR].

Further studies indicate that curcumin and helenalin, which contain alpha, beta unsaturated carbonyl group, but not resveratrol (with no unsaturated carbonyl group, XREF_FIG), inhibit TLR4 activation by interfering with receptor dimerization [XREF_BIBR] (XREF_FIG).

Curcumin, helenalin, and cinnamaldehyde with alpha, beta unsaturated carbonyl groups, or sulforaphane with an isothiocyanate group, inhibit TLR4 activation by interfering with cysteine residue mediated receptor dimerization, while resveratrol, with no unsaturated carbonyl group, did not.

Such conclusion was further supported by the result that curcumin inhibits ligand independent dimerization of constitutively active TLR4.

Curcumin, helenalin, cinnamaldehyde and sulforaphane, containing alpha, beta unsaturated carbonyl or isothiocyanate group, respectively, that are known to interact with free SH groups in cysteine residues, but not resveratrol (with no unsaturated carbonyl group), inhibit TLR4 activation by interfering with TLR4 receptor dimerization.

Furthermore, the suppressive effect of resveratrol on LPS induced NF-kappaB activation was abolished in TRIF deficient mouse embryonic fibroblasts, but not in MyD88 deficient macrophages (XREF_FIG), suggesting that resveratrol specifically inhibits MyD88 independent signaling pathways downstream of TLR3 and TLR4.

Furthermore, the suppressive effect of resveratrol on LPS induced NF-kappaB activation was abolished in TRIF deficient mouse embryonic fibroblasts, but not in MyD88 deficient macrophages (XREF_FIG), suggesting that resveratrol specifically inhibits MyD88 independent signaling pathways downstream of TLR3 and TLR4.

In contrast, resveratrol, EGCG, luteolin, and structural analogs of luteolin specifically inhibit TLR3 and TLR4 signaling by targeting TANK binding kinase 1 (TBK1) and receptor interacting protein 1 (RIP1) in Toll/IL -1 receptor domain containing adaptor inducing IFN-beta (TRIF) complex.

Together, these results demonstrate that resveratrol specifically inhibits TLR3 and TLR4 signaling pathway by targeting TBK1 and RIP1 in TRIF complex.

In contrast, resveratrol, EGCG, luteolin and structural analogs of luteolin, specifically inhibit TLR3 and TLR4 signaling by targeting TBK1 and RIP1 in TRIF complex.

Together, these results demonstrate that resveratrol specifically inhibits TLR3 and TLR4 signaling pathway by targeting TBK1 and RIP1 in TRIF complex.

In contrast, resveratrol, EGCG, luteolin and structural analogs of luteolin, specifically inhibit TLR3 and TLR4 signaling by targeting TBK1 and RIP1 in TRIF complex.

In contrast, resveratrol, EGCG, luteolin, and structural analogs of luteolin specifically inhibit TLR3 and TLR4 signaling by targeting TANK binding kinase 1 (TBK1) and receptor interacting protein 1 (RIP1) in Toll/IL -1 receptor domain containing adaptor inducing IFN-beta (TRIF) complex.

In further delineating the target of resveratrol, it was found that resveratrol inhibited the kinase activity of TBK1 and the NF-kappaB activation induced by RIP1 [XREF_BIBR] (XREF_FIG).

Curcumin, helenalin, and cinnamaldehyde with alpha, beta unsaturated carbonyl groups, or sulforaphane with an isothiocyanate group, inhibit TLR4 activation by interfering with cysteine residue mediated receptor dimerization, while resveratrol, with no unsaturated carbonyl group, did not.

Curcumin, helenalin, cinnamaldehyde and sulforaphane, containing alpha, beta unsaturated carbonyl or isothiocyanate group, respectively, that are known to interact with free SH groups in cysteine residues, but not resveratrol (with no unsaturated carbonyl group), inhibit TLR4 activation by interfering with TLR4 receptor dimerization.

Additional studies showed that EGCG, luteolin and structural analogs of luteolin such as quercetin, chrysin, and eriodictyol, also inhibited TRIF signaling pathway by targeting TBK1 kinase [XREF_BIBR, XREF_BIBR].

In contrast, resveratrol, EGCG, luteolin and structural analogs of luteolin, specifically inhibit TLR3 and TLR4 signaling by targeting TBK1 and RIP1 in TRIF complex.

In contrast, resveratrol, EGCG, luteolin, and structural analogs of luteolin specifically inhibit TLR3 and TLR4 signaling by targeting TANK binding kinase 1 (TBK1) and receptor interacting protein 1 (RIP1) in Toll/IL -1 receptor domain containing adaptor inducing IFN-beta (TRIF) complex.

Insulin induced mRNA or protein expressions of cell death signaling markers such as cleaved PARP, p53, and Bax, as well as the ER stress markers such as p-eIF2alpha, ATF4, CHOP, sXBP1, and p-IRE1 were significantly attenuated by melatonin treatment.

Melatonin Attenuates the Activation of ASK1 under IR Condition.

On the other hand, the decreased cell viability by insulin stimulation was recovered when treated with melatonin.

In addition, immunofluorescence analysis confirmed that p-IRE1 induced by insulin stimulation was significantly suppressed by melatonin treatment (XREF_FIG C).

Insulin induced mRNA or protein expressions of cell death signaling markers such as cleaved PARP, p53, and Bax, as well as the ER stress markers such as p-eIF2alpha, ATF4, CHOP, sXBP1, and p-IRE1 were significantly attenuated by melatonin treatment.

Insulin induced mRNA or protein expressions of cell death signaling markers such as cleaved PARP, p53, and Bax, as well as the ER stress markers such as p-eIF2alpha, ATF4, CHOP, sXBP1, and p-IRE1 were significantly attenuated by melatonin treatment.

Insulin induced mRNA or protein expressions of cell death signaling markers such as cleaved PARP, p53, and Bax, as well as the ER stress markers such as p-eIF2alpha, ATF4, CHOP, sXBP1, and p-IRE1 were significantly attenuated by melatonin treatment.

EGCG also markedly inhibited the activation of Stat3 in YCU-H891 (carcinoma of the hypopharynx) cells [XREF_BIBR] and BT-474 (human breast cancer cell) cells [XREF_BIBR].

Curcumin is a potent inhibitor of COX-2.

Curcumin was shown to inhibit constitutive and IL-6-induced Stat3 activation in human multiple myeloma cells [XREF_BIBR].

Resveratrol dose-dependently prevented both COX-2 induction and PGE (2) production in bFGF stimulated fibroblasts [XREF_BIBR].

Resveratrol suppressed NF-kappaB-regulated gene products (COX-2, MMP-3, MMP-9, and VEGF), inhibited anti-apoptosis (Bcl-2, and Bcl-xL) [XREF_BIBR].

Treatment of quercetin down-regulated the antiapoptotic proteins Bcl-2 and Bcl-xL and also up-regulated the proapoptotic proteins Bax and caspase-3 in prostatic PC-3 carcinoma cells [XREF_BIBR].

Treatment of quercetin down-regulated the antiapoptotic proteins Bcl-2 and Bcl-xL and also up-regulated the proapoptotic proteins Bax and caspase-3 in prostatic PC-3 carcinoma cells [XREF_BIBR].

Antivirals such as camostat mesylate (inhibitor of TMPRSS2), chloroquine and hydroxychloroquine (inhibitor of endocytosis), lopinavir and darunavir (inhibitor of 3-chymotrypsin-like protease) or ribavirin, remdesivir, favipiravir (inhibitor of RNA dependent RNA polymerase), or prednisolone should be restricted to controlled or randomized trials such as the worldwide WHO cosponsored Solidarity Trial (https://www.who.int/emergencies/diseases/novel-coronavirus-2019/global-research-on-novel-coronavirus-2019-ncov/solidarity-clinical-trial-for-covid-19-treatments).

Antivirals such as camostat mesylate (inhibitor of TMPRSS2), chloroquine and hydroxychloroquine (inhibitor of endocytosis), lopinavir and darunavir (inhibitor of 3-chymotrypsin-like protease) or ribavirin, remdesivir, favipiravir (inhibitor of RNA dependent RNA polymerase), or prednisolone should be restricted to controlled or randomized trials such as the worldwide WHO cosponsored Solidarity Trial (https://www.who.int/emergencies/diseases/novel-coronavirus-2019/global-research-on-novel-coronavirus-2019-ncov/solidarity-clinical-trial-for-covid-19-treatments).

Antivirals such as camostat mesylate (inhibitor of TMPRSS2), chloroquine/hydroxychloroquine (inhibitor of endocytosis), lopinavir/darunavir (inhibitor of 3‑chymotrypsin-like protease) or ribavirin, remdesivir, favipiravir (inhibitor of RNA-dependent RNA polymerase), or prednisolone should be restricted to controlled or randomized trials such as the worldwide WHO-cosponsored Solidarity Trial (https://www.who.int/emergencies/diseases/novel-coronavirus-2019/global-research-on-novel-coronavirus-2019-ncov/solidarity-clinical-trial-for-covid-19-treatments).

Antivirals such as camostat mesylate (inhibitor of TMPRSS2), chloroquine and hydroxychloroquine (inhibitor of endocytosis), lopinavir and darunavir (inhibitor of 3-chymotrypsin-like protease) or ribavirin, remdesivir, favipiravir (inhibitor of RNA dependent RNA polymerase), or prednisolone should be restricted to controlled or randomized trials such as the worldwide WHO cosponsored Solidarity Trial (https://www.who.int/emergencies/diseases/novel-coronavirus-2019/global-research-on-novel-coronavirus-2019-ncov/solidarity-clinical-trial-for-covid-19-treatments).

Antivirals such as camostat mesylate (inhibitor of TMPRSS2), chloroquine and hydroxychloroquine (inhibitor of endocytosis), lopinavir and darunavir (inhibitor of 3-chymotrypsin-like protease) or ribavirin, remdesivir, favipiravir (inhibitor of RNA dependent RNA polymerase), or prednisolone should be restricted to controlled or randomized trials such as the worldwide WHO cosponsored Solidarity Trial (https://www.who.int/emergencies/diseases/novel-coronavirus-2019/global-research-on-novel-coronavirus-2019-ncov/solidarity-clinical-trial-for-covid-19-treatments).

Antivirals such as camostat mesylate (inhibitor of TMPRSS2), chloroquine/hydroxychloroquine (inhibitor of endocytosis), lopinavir/darunavir (inhibitor of 3‑chymotrypsin-like protease) or ribavirin, remdesivir, favipiravir (inhibitor of RNA-dependent RNA polymerase), or prednisolone should be restricted to controlled or randomized trials such as the worldwide WHO-cosponsored Solidarity Trial (https://www.who.int/emergencies/diseases/novel-coronavirus-2019/global-research-on-novel-coronavirus-2019-ncov/solidarity-clinical-trial-for-covid-19-treatments).

Antivirals such as camostat mesylate (inhibitor of TMPRSS2), chloroquine and hydroxychloroquine (inhibitor of endocytosis), lopinavir and darunavir (inhibitor of 3-chymotrypsin-like protease) or ribavirin, remdesivir, favipiravir (inhibitor of RNA dependent RNA polymerase), or prednisolone should be restricted to controlled or randomized trials such as the worldwide WHO cosponsored Solidarity Trial (https://www.who.int/emergencies/diseases/novel-coronavirus-2019/global-research-on-novel-coronavirus-2019-ncov/solidarity-clinical-trial-for-covid-19-treatments).

Antivirals such as camostat mesylate (inhibitor of TMPRSS2), chloroquine/ hydroxychloroquine (inhibitor of endocytosis), lopinavir/darunavir (inhibitor of 3-chymotrypsin-like protease) or ribavirin, remdesivir, favipiravir (inhibitor of RNA-dependent RNA polymerase), or prednisolone should be restricted to controlled or randomized trials such as the worldwide WHO-cosponsored Solidarity Trial (https://www.

Antivirals such as camostat mesylate (inhibitor of TMPRSS2), chloroquine and hydroxychloroquine (inhibitor of endocytosis), lopinavir and darunavir (inhibitor of 3-chymotrypsin-like protease) or ribavirin, remdesivir, favipiravir (inhibitor of RNA dependent RNA polymerase), or prednisolone should be restricted to controlled or randomized trials such as the worldwide WHO cosponsored Solidarity Trial (https://www.who.int/emergencies/diseases/novel-coronavirus-2019/global-research-on-novel-coronavirus-2019-ncov/solidarity-clinical-trial-for-covid-19-treatments).

Antivirals such as camostat mesylate (inhibitor of TMPRSS2), chloroquine/hydroxychloroquine (inhibitor of endocytosis), lopinavir/darunavir (inhibitor of 3‑chymotrypsin-like protease) or ribavirin, remdesivir, favipiravir (inhibitor of RNA-dependent RNA polymerase), or prednisolone should be restricted to controlled or randomized trials such as the worldwide WHO-cosponsored Solidarity Trial (https://www.who.int/emergencies/diseases/novel-coronavirus-2019/global-research-on-novel-coronavirus-2019-ncov/solidarity-clinical-trial-for-covid-19-treatments).