姜黄素调控自噬在抑制HPV阳性宫颈癌细胞的体外研究

Curcumin regulated mitochondrial autophagy in the inhibition of HPV-positive cervical cancer cells

目的研究姜黄素在体外应用于诱导HPV阳性宫颈癌细胞凋亡与自噬的影响。方法采用MTT检测法和流式细胞仪检测姜黄素(10,20,30,40μmol/L)对HPV阳性宫颈癌SiHa细胞增殖和凋亡的影响;采用自噬抑制剂3-MA与姜黄素联用,观察自噬对姜黄素抑制HPV阳性宫颈癌细胞增殖和诱导细胞凋亡的影响;分光光度法检测SiHa细胞Caspase-3活性;应用ELISA检测姜黄素和(或)自噬抑制剂3-MA对宫颈癌SiHa细胞内Bcl-2、Bax和Beclin-1表达的影响。结果姜黄素对HPV阳性的宫颈癌SiHa细胞的生长具有抑制作用,而且这种抑制作用呈浓度依赖性和时间依赖性;而姜黄素与3-MA联用后,对HPV阳性宫颈癌SiHa细胞诱导凋亡的作用明显下降;与对照组相比,姜黄素组细胞Caspase-3活性明显提高(P<0.01)、Bax和Beclin-1的表达均明显升高(P<0.01),而Bcl-2的表达明显降低(P<0.01);与单独应用姜黄素组相比,姜黄素与3-MA联用组细胞Caspase-3活性和Bax表达均明显下降(P<0.01),而Bcl-2的表达明显增加(P<0.01)。结论姜黄素能有效抑制HPV阳性宫颈癌SiHa细胞的增殖,可以提高细胞Caspase-3活性、促进促凋亡因子Bax的表达升高和抑制凋亡因子Bcl-2表达下调,同时促进自噬相关蛋白Beclin 1的表达上调,从而诱导HPV阳性宫颈癌SiHa细胞发生凋亡和自噬性细胞死亡。

Objective To study the effect of curcumin on the apoptosis and autophagy of HPV-positive cervical cancer cells in vitro.Methods The effects of curcumin（10,20,30,40 μmol/L）on the proliferation and apoptosis of HPV-positive cervical cancer SiHa cells were detected by MTT assay and flow cytometry;autophagy inhibitor 3-MA was used in combination with curcumin to observe the effect of autophagy on curcumin inhibition of HPV positive cervical cancer cell proliferation and induction of apoptosis;the expression of Caspase-3 in SiHa cells was detected by spectrophotometry,and the effects of curcumin and/or autophagy inhibitors（3-MA）on the expression of Bcl-2 and Bax in cervical cancer SiHa cells were detected by ELISA.Results Curcumin inhibited the growth of HPV-positive cervical cancer SiHa cells,and the inhibitory effect was in a dose-dependent and time-dependent manner.After curcumin combined with 3-MA,the effect of curcumin on the HPV-positive cervical cancer SiHa The effect of cell apoptosis was significantly decreased.Compared with the control group,the expression of Caspase-3,Bax and Beclin-1 in the curcumin group was significantly higher than that in the control group（P〈0.01）,while the expression of Bcl-2 was significantly decreased（P〈0.01）（P〈0.01）,and the expression of Bcl-2 was significantly higher in the combination of curcumin and 3-MA group（P〈0.01）,and the expression of Caspase-3,Bax and Beclin-1 increased（P〈0.01）.Conclusion Curcumin can effectively inhibit the proliferation of HPV-positive cervical cancer SiHa cells,can promote the expression of pro-apoptotic factors Bax,Caspase-3 and inhibit the down-regulation of apoptosis factor Bcl-2 expression,while promoting the expression of autophagy-related protein Beclin 1 Upregulation,which induced HPV-positive cervical cancer SiHa cells apoptosis and autophagic cell death.