噻可拉林协同顺铂促进宫颈癌c4-1及Hela细胞凋亡机制的研究

Mechanism of thiocoraline combination with cisplatin on promoting apoptosis of cervical cancer cell line c4-1 and Hela

目的:探讨噻可拉林协同顺铂促进宫颈癌c4-1、Hela细胞凋亡的机制。方法:用不同浓度噻可拉林和顺铂单独或联合作用于体外培养的c4-1、Hela细胞,MTT法检测c4-1、Hela细胞生长抑制率,Hoechst 33342和AO-EB染色法检测细胞凋亡,Western blot检测细胞Bax、Bcl2、p53、p21蛋白的表达,流式细胞术检测细胞增殖周期。结果:c4-1及Hela细胞经不同浓度的噻可拉林和顺铂单独或联合作用后,细胞的体外增殖活力被明显抑制,呈剂量依赖关系,且在联合使用噻可拉林(10nmol/L):顺铂(4μmol/L)用量时抑制效果最明显(P<0.05);AO-EB染色法显示,噻可拉林和顺铂联合使用,细胞凋亡更明显。Western blot则显示,Bax、p53、p21表达明显上调,而Bcl2的表达则明显下调。结论:噻可拉林能协同顺铂上调Bax、p53、p21的表达和下调Bcl2的表达,抑制宫颈癌c4-1、Hela细胞增殖同时促进细胞凋亡。

Objective： To investigate the apoptosis by thiocoraline in combination with cisplatin on cervical cell line c4-1 and Hela and its mechanism. Methods： Different concentrations of thiocoraline and cisplatin alone or in combination used for cultured c4-1and Hela cells. The inhibition of cell proliferation was detected by MTT assay,and the apoptosis was detected by Hoechst 33342 and AO-EB staining. The expressions of the protein Bax,Bcl2,p53,p21 were detected by Western blot. The cell cycle of c4-1 and Hela cells were detected by Flow cytometry. Results： The cell proliferation activity of c4-1 and He La cells were inhibited obviously by different concentrations of thiocoraline and cisplatin alone or in combination. The inhibition effect was in a dose-dependent manner. Meanwhile,when the concentrations of thiocoraline and cisplatin were 10 nmol / L and 4 μmol / L,respectivity the inhibitory effect was most significant（ P〈0. 05）; Hoechst 33342 and AO-EB staining showed that the cell apoptosis effect was more obvious on thiocoraline in combination with cisplatin. The Western blot showed that the expressions of the protein Bax,p53,p21 were significantly up-regulated,and the Bcl2 was significantly down-regulated. Conclusion： Thiocoraline combination with cisplatin could up-regulate the expression of the protein Bax,p53,p21 and down-regulation the expression of Bcl2 to inhibition the proliferation and promotion apoptosis of cervical cell line c4-1 and Hela.