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Curcumin Inhibits Viability of Clear Cell Renal Cell Carcinoma by Down-Regulating adamts18 gene Methylation through NF-κB and AKT Signaling Pathway

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Chinese Journal of Integrative Medicine 2022年第5期28卷 419-424页

作者： XU Ben PENG Yi-ji ZHU Wei-jie Department of Urology Peking University First Hospital and Institute of UrologyPeking UniversityNational Urological Cancer CenterBeijing(100034)China

Objective:To investigate the effect of curcumin on viability of clear cell renal cell carcinoma(cc RCC)and analyze its possible mechanism.Methods:In cell lines of A498 and 786-O,the effects of curcumin(1.25,2.5,5 and 10μmol/L)on the viability of cc RCC were analyzed at 24,48 and 72 h by MTT assay.The protein expression levels of adamts18 gene,p65,phosphorylation p65(pp65),AKT,phosphorylation AKT(p AKT)and matrix metallopeptidase 2(MMP-2)before and after curcumin(10μmol/L)treatment were examined by Western blotting.Real-time PCR and methylation specific PCR(MSP)were applied to analyze the expression and methylation level of adamts18 gene before and after curcumin treatment(10μmol/L).Results:Curcumin significantly inhibited the viability of A498 and 786-O cell lines in a dose-and time-dependent manner(Padamts18 gene expression with down-regulation of adamts18 gene methylation was reflected after curcumin treatment,accompanied by down-regulation of nuclear factorκB(NF-κk B)related protein(p65 and pp65),AKT related protein(AKT and p AKT),and NF-κB/AKT common related protein MMP-2.With adamts18 gene overexpressed,the expression levels of p65,AKT and MMP2 were downregulated,of which were conversely up-regulated in silenced adamts18(sh-adamts18).The expression of pp65,p AKT and MMP2 in sh-adamts18 was down-regulated after being treated with PDTC(NF-κB inhibitor)and LY294002(AKT inhibitor).Conclusion:Curcumin could inhibit the viability of cc RCC by down-regulating adamts18 gene methylation though NF-κB and AKT signaling pathway.

关键词： curcumin clear cell renal cell carcinoma adamts18 gene methylation signaling pathway

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Curcumin Inhibits Proliferation of Renal Cell Carcinoma in vitro and in vivo by Regulating miR-148/adamts18 through Suppressing Autophagy

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Chinese Journal of Integrative Medicine 2023年第8期29卷 699-706页

作者： XU Ben YUAN Chang-wei ZHANG Jia-en Depatment of Urology Peking University First HospitalBeijing(100034)China

Objective: To explore the effect of curcumin on the proliferation of renal cell carcinoma and analyze its regulation mechanism. Methods: In RCC cell lines of A498 and 786-O, the effects of curcumin(2.5, 5, 10 μmo/L) on the proliferation were analyzed by Annexin V+PI staining. Besides, A498 was inoculated into nude mice to establish tumorigenic models, and the model mice were treated with different concentrations of curcumin(100, 200, and 400 mg/kg), once daily for 30 days. Then the tumor diameter was measured, the tumor cells were observed by hematoxylin-eosin staining, and the protein expressions of miR-148 and adamts18were detected by immunohistochemistry. In vitro, after transfection of miR-148 mimics, miR-148 inhibitor or si-adamts18 in cell lines, the expression of adamts18 was examined by Western blotting and the cell survival rate was analyzed using MTT. Subsequently, Western blot analysis was again used to examine the autophagy phenomenon by measuring the relative expression level of LC3-Ⅱ/LC3-Ⅰ;autophagy-associated genes,including those of Beclin-1 and ATG5, were also examined when miR-148 was silenced in both cell lines with curcumin treatment. Results: Curcumin could inhibit the proliferation of RCC in cell lines and nude mice. The expressions of miR-148 and adamts18 were upregulated after curcumin treatment both in vitro and in vivo(Padamts18 upregulated. However,si-adamts18 treatment or miR-148 inhibitor reversed these results, that is, both of them promoted the cell survival rate. Conclusion: Curcumin can inhibit the proliferation of renal cell carcinoma by regulating the miR-148/adamts18 axis through the suppression of autophagy in vitro and in vivo. There may exist a positive feedback loop between mi R-148 and adamts18 gene in RCC.

关键词： curcumin renal cell carcinoma MiR-148 adamts18 gene autophagy

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