姜黄素对肾癌ACHN细胞增殖与凋亡的影响及作用机制

Effects and mechanism of Curcumin on proliferation and apoptosis of renal cell carcinoma ACHN cells

目的研究姜黄素对肾癌ACHN细胞增殖与凋亡的影响,并探讨其中的作用机制。方法不同浓度(0、10、20、40、80μmol/L)姜黄素作用于ACHN细胞不同时间(12、24、48 h)后,利用CCK-8检测细胞增殖活性的改变;应用Annexin-V/PI双标染色后上流式细胞仪检测细胞凋亡变化;Real-time PCR测定Notch1基因表达量的变化;免疫印迹法检测凋亡相关蛋白Bcl-2、Bax、Caspase-3和Notch蛋白表达情况。结果CCK-8结果表明,姜黄素作用后ACHN细胞增殖速度减慢(P<0.05),且抑制作用呈时间及浓度依赖性;流式检测结果显示姜黄素作用后ACHN细胞凋亡增加(P<0.05);Real-time PCR检测结果显示Notch1基因表达增加,且表达量呈时间及浓度依赖性(P<0.05);Western blot检测发现随着姜黄素浓度的增加,Bcl-2蛋白表达下降,Bax、Caspase-3、Notch1蛋白表达增加。结论姜黄素能够抑制肾癌ACHN细胞的增殖并诱导其凋亡,其过程可能通过调节Notch1基因的表达,抑制抗凋亡因子Bcl-2蛋白表达,促进凋亡因子Bax蛋白表达,从而激活促凋亡因子Caspase-3,继而引发凋亡反应。

Objective To investigate the effects of Curcumin on the proliferation and apoptosis of renal cell carcinoma ACHN cells,and to probe the mechanism.Methods ACHN cells were treated with different concentrations of Curcumin for different duration.CCK-8 was used to measure the cell proliferation activity.Apoptosis change was measured by flow cytometry after Annexin V/PI double labeling.Real-time PCR was used to analyze the changes in Notch1 gene mRNA.The protein level of Bcl-2 Caspase-3 and Notch1 was measured by Western blot.Results CCK-8 results showed that the proliferation rate of ACHN cells significantly reduced after Curcumin exposure compared with the control group(P<0.05),and the inhibition rate was time-and dose-dependent.Flow cytometry results showed that the apoptosis of ACHN cells was significantly increased after Curcumin treatment(P<0.05).Notch1 detection showed Notch1 was significantly increased in a time-and dose-dependent manner.Western blot analysis showed that the expression of Bcl-2 decreased and Bax,Caspase-3 and Notch1 increased positively with the concentration of Curcumin.Conclusion Curcumin can inhibit the proliferation of ACHN cells and induce apoptosis,the mechanism is probably correlated with the expression of Notch1 gene and the protein expression of Caspase-3,Bcl-2 and Bax.