Xu Ben, Peng Yi-Ji, Zhu Wei-Jie
Department of Urology, Peking University First Hospital and Institute of Urology, Peking University, National Urological Cancer Center, Beijing, 100034, China.
Chin J Integr Med. 2022 May;28(5):419-424. doi: 10.1007/s11655-021-3445-z. Epub 2021 May 17.
To investigate the effect of curcumin on viability of clear cell renal cell carcinoma (ccRCC) and analyze its possible mechanism.
In cell lines of A498 and 786-O, the effects of curcumin (1.25, 2.5, 5 and 10 μ mol/L) on the viability of ccRCC 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 (pAKT) and matrix metallopeptidase 2 (MMP-2) before and after curcumin (10 μ mol/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 (10 μ mol/L).
Curcumin significantly inhibited the viability of A498 and 786-O cell lines in a dose- and time-dependent manner (P<0.01). Up-regulation of ADAMTS18 gene expression with down-regulation of ADAMTS18 gene methylation was reflected after curcumin treatment, accompanied by down-regulation of nuclear factor κ B (NF-κ kB) related protein (p65 and pp65), AKT related protein (AKT and pAKT), and NF-κ B/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, pAKT and MMP2 in sh-ADAMTS18 was down-regulated after being treated with PDTC (NF-κ B inhibitor) and LY294002 (AKT inhibitor).
Curcumin could inhibit the viability of ccRCC by down-regulating ADAMTS18 gene methylation though NF-κ B and AKT signaling pathway.
探讨姜黄素对肾透明细胞癌（ccRCC）细胞活力的影响并分析其可能机制。
采用MTT法分析姜黄素（1.25、2.5、5和10 μmol/L）在24、48和72小时对A498和786 - O细胞系中ccRCC细胞活力的影响。用蛋白质免疫印迹法检测姜黄素（10 μmol/L）处理前后ADAMTS18基因、p65、磷酸化p65（pp65）、AKT、磷酸化AKT（pAKT）和基质金属蛋白酶2（MMP - 2）的蛋白表达水平。应用实时荧光定量PCR和甲基化特异性PCR（MSP）分析姜黄素（10 μmol/L）处理前后ADAMTS18基因的表达及甲基化水平。
姜黄素能以剂量和时间依赖性方式显著抑制A498和786 - O细胞系的活力（P<0.01）。姜黄素处理后可使ADAMTS18基因表达上调，ADAMTS18基因甲基化下调，同时核因子κB（NF - κB）相关蛋白（p65和pp65）、AKT相关蛋白（AKT和pAKT）以及NF - κB/AKT共同相关蛋白MMP - 2表达下调。过表达ADAMTS18基因后，p65、AKT和MMP2表达下调，而在ADAMTS18基因沉默（sh - ADAMTS18）时则相反上调。用PDTC（NF - κB抑制剂）和LY294002（AKT抑制剂）处理sh - ADAMTS18后，pp65、pAKT和MMP2的表达下调。
姜黄素可通过下调ADAMTS18基因甲基化，经NF - κB和AKT信号通路抑制ccRCC细胞活力。
