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目的研究异甘草素(ISL)抑制宫颈癌Hela细胞增殖作用及促进细胞凋亡的作用机制。方法采用0.025、0.050、0.100、0.200、0.300、0.400mg/mL浓度ISL处理HeLa细胞,采用四甲基偶氮唑盐(MTT)比色法检测HeLa细胞增殖;采用流式细胞仪检测HeLa细胞凋亡情况;应用罗丹明123染色法检测HeLa细胞线粒体跨膜电位的变化;采用RT-PCR检测6种不同浓度ISL对Bcl-2、P53、P21、E6mRNA基因表达的影响。结果 (1)MTT检测结果显示:随着ISL浓度增加,对不同时间段HeLa细胞的抑制作用明显增加(P<0.05);(2)流式细胞仪检测:相比于空白对照组,随着ISL浓度逐渐增加,Hela细胞凋亡率明显升高(P<0.01),且凋亡率呈剂量依赖性;(3)罗丹明123荧光染色检测结果:随着ISL浓度逐渐增加,细胞线粒体膜电位下降(P<0.01);(4)RT-PCR结果显示:随着ISL浓度逐渐增加,E6、Bcl-2mRNA表达水平明显降低(P<0.05),而P53、p21mRNA表达水平越升高(P<0.05)。结论 ISL通过下调E6、Bcl-2的表达和上调P21、P53基因的表达来抑制宫颈癌细胞的增殖。
Objective To study the mechanism of isoliquiritigenin (ISL) inhibiting cervical cancer Hela cell proliferation and promoting apoptosis. Methods HeLa cells were treated with 0.025,0.050,0.100,0.200,0.300,0.400mg / mL ISL, and the proliferation of HeLa cells was detected by MTT assay. The apoptosis of HeLa cells was detected by flow cytometry The changes of mitochondrial transmembrane potential of HeLa cells were detected by rhodamine 123 staining. The effects of six different concentrations of ISL on the gene expression of Bcl-2, P53, P21 and E6 were detected by RT-PCR. Results (1) The results of MTT assay showed that with the increase of ISL concentration, the inhibitory effect on HeLa cells was significantly increased at different time points (P <0.05). (2) Flow cytometry: Compared with the blank control group, (3) The results of rhodamine 123 fluorescence staining showed that with the increasing of ISL concentration, the mitochondrial membrane potential of Hela cells increased significantly (P <0.01), and the apoptosis rate of Hela cells increased in a dose- (P <0.01). (4) The results of RT-PCR showed that the expressions of E6 and Bcl-2 mRNA were significantly decreased and the expressions of P53 and p21 mRNA increased with the increasing of ISL concentration (P < 0.05). Conclusion ISL inhibits the proliferation of cervical cancer cells by down-regulating the expressions of E6 and Bcl-2 and up-regulating the expressions of P21 and P53 genes.