复方山慈菇醇提物对人乳腺癌MDA-MB-231细胞增殖与凋亡的影响

Effects of compound extract of pseudobulbus cremastrae seu pleiones on proliferation and apoptosis of human breast cancer MDA-MB-231 cells

目的:探讨复方山慈菇醇提物对人三阴性乳腺癌(TNBC)MDA-MB-231细胞增殖与凋亡的影响。方法:将人乳腺癌MDA-MB-231细胞分为溶剂对照组和复方山慈菇醇提物组,0.1%DMSO或不同浓度的复方山慈菇醇提物作用于MDA-MB-231细胞24 h、48 h后,CCK-8法检测细胞活性;Hoechst33342染色法观察细胞形态学变化;Annexin V-APC/7-AAD双染色流式分析检测细胞凋亡率;Western blot检测凋亡相关蛋白Bcl-2、Bax和Caspase-3的表达。结果:复方山慈菇醇提物可浓度和时间依赖性地抑制MDA-MB-231细胞的增殖(P<0.05),24 h、48 h的IC50分别为0.51 mg生药/mL和0.26 mg生药/mL;0.25 mg生药/mL、0.5 mg生药/mL、1.0 mg生药/mL复方山慈菇醇提物处理24 h后,Hoechst33342染色显示细胞出现凋亡样变化;流式细胞术检测结果表明,与溶剂对照组比较,复方山慈菇醇提物可浓度依赖性地增加MDAMB-231细胞凋亡率(P<0.05);Western blot法显示,与溶剂对照相比,0.5 mg生药/mL复方山慈菇醇提物组Bax、Caspase-3蛋白表达升高,Bcl-2蛋白表达下降(均P<0.05)。结论:复方山慈菇可体外抑制人乳腺癌MDA-MB-231细胞的增殖,机制可能与其下调Bcl-2及上调Bax、Caspase-3蛋白表达而诱导凋亡有关。

Objective: To explore the effect of compound extract of pseudobulbus cremastrae seu pleiones on proliferation and apoptosis of human triple negative breast cancer(TNBC)MDA-MB-231 cells.Methods: Human breast cancer MDA-MB-231 cells were divided into solvent control group and compound extract of pseudobulbus cremastrae seu pleiones group.After MDA-MB-231 cells were treated with 0.1% DMSO or different concentrations of compound extract of pseudobulbus cremastrae seu pleiones for 24 h and 48 h,the cell viability was detected by CCK-8 method,the cell morphological changes were observed by Hoechst 33342 staining,and the apoptosis rate was detected by Annexin V-APC/7-AAD double staining flow analysis.Western blot was used to detect the expression of apoptosis-related proteins Bcl-2,Bax,and Caspase-3.Results: The compound extract of pseudobulbus cremastrae seu pleiones could inhibit the proliferation of MDA-MB-231 cells in a concentration-and time-dependent manner(P<0.05).The IC50 of 24 h and 48 h were 0.51 mg/mL and 0.26 mg/mL.After treated with 0.25 mg/mL,0.5 mg/mL,1.0 mg/mL compound extract of pseudobulbus cremastrae seu pleiones for 24 h,Hoechst 33342 staining showed MDA-MB-231 cells had apoptosis-like changes.The results of flow cytometry showed that compared with the solvent control group,the compound extract of pseudobulbus cremastrae seu pleiones could increase the apoptosis rate of MDA-MB-231 cells in a concentration-dependent manner(P<0.05).Western blot showed that compared with the solvent control,the expression of Bax and Caspase-3 protein were increased and the expression of Bcl-2 protein was decreased in 0.5 mg/mL compound extract group of pseudobulbus cremastrae seu pleiones(all P<0.05).Conclusion: Compound pseudobulbus cremastrae seu pleiones can inhibit the proliferation of human breast cancer MDA-MB-231 cells in vitro,and the mechanism may be related to induce apoptosis by down-regulation of Bcl-2 expression and up-regulation of Bax and Caspase-3 protein expression.