姜黄素通过调节TLR4/NF⁃κB信号通路抑制肾细胞癌恶性表型的研究

An experimental study of curcumin inhibiting the malignant phenotype of renal cell carcinoma by regulating the TLR4/NF-κB signaling pathway

目的探究姜黄素通过调节Toll样受体4/核转录因子-κB(TLR4/NF-κB)信号通路,抑制人肾细胞癌(RCC)恶性表型。方法将体外培养的HK-2、786-0、ACHN细胞株分为对照组、姜黄素组、TLR4/NF-κB信号通路抑制剂组(瑞沙托维,TAK-242组),其中姜黄素组依据不同干预剂量分为姜黄素10、20、40μmol/L亚组。CCK-8法、Annexin V-FITC/碘化丙啶染色、划痕实验、Transwell小室实验分别检测细胞增殖、凋亡、迁移、侵袭能力;ELISA法检测细胞白介素-1β(IL-1β)、白介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)含量;蛋白印迹法检测TLR4、NF-κB p65、B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X的蛋白质(Bax)、基质金属蛋白酶9(MMP-9)的表达。结果姜黄素在40μmol/L以下浓度时不存在毒性作用;在RCC细胞株786-0、ACHN中,与对照组相比,姜黄素10、20、40μmol/L亚组能够明显抑制细胞增殖、迁移和侵袭,促进细胞凋亡;IL-1β、IL-6和TNF-α、TLR4、NF-κB p65、Bcl-2、MMP-9水平降低,Bax水平升高,差异均有统计学意义(均P<0.05)。结论姜黄素可通过抑制TLR4/NF-κB信号通路,抑制RCC细胞的增殖、迁移和侵袭,并促进细胞凋亡。

Objective To explore curcumin inhibiting the malignant phenotype of human renal cell carcinoma(RCC)by regulating the Toll⁃like receptor 4/nuclear factor⁃κB(TLR4/NF⁃κB)signaling pathway.Method The HK⁃2,786⁃0,and ACHN cell lines cultured in vitro were divided into control group,curcumin group,TLR4/NF⁃κB signaling pathway inhibitor group(TAK⁃242 group).The curcumin group was divided into curcumin 10,20,40μmol/L subgroup.CCK⁃8 method,Annexin V⁃FITC/PI staining,scratch test,and Transwell chamber test were used to detect ability of cell proliferation,apoptosis,migration and invasion;ELISA method was used to detect cell interleukin⁃1β(IL⁃1β),interleukin⁃6(IL⁃6)and tumor necrosis factor⁃α(TNF⁃α)content;Western blotting method was used to detect the expression of TLR4,NF⁃κB p65,B cell lymphoma⁃2(Bcl⁃2),BCL2⁃Associated X(Bax),and matrix metalloprotein⁃9(MMP⁃9).Result Curcumin had no toxic effect at concentrations below 40μmol/L;in renal cancer cell lines 786⁃0 and ACHN,Compared with the control group,curcumin 10,20,and 40μmol/L subgroups can significantly inhibit cell proliferation,migration and invasion,and promote cell apoptosis;IL⁃1β,IL⁃6 and TNF⁃α,TLR4,NF⁃κB p65,Bcl⁃2,MMP⁃9 levels decrease,and Bax levels increase,The differences are statistically significant(P<0.05).Conclusion Curcumin can inhibit the proliferation,migration and invasion of renal cancer cells and promote cell apoptosis by inhibiting the TLR4/NF⁃κB signaling pathway.