大黄酸衍生物H对MDA-MB-231细胞增殖的抑制作用

Inhibitory effect of Rhein derivative H on the proliferation of MDA-MB-231 cells

目的探究大黄酸衍生物H(文中简称H)对人三阴性乳腺癌细胞MDA-MB-231增殖凋亡的影响及其机制。方法以MDA-MB-231细胞为研究对象,分为无H处理的对照组和低、中、高剂量实验组(1,2,4μmol·L^-1),用流式细胞术检测细胞凋亡,蛋白免疫印迹法检测H对凋亡相关蛋白B细胞淋巴瘤/白血病-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、聚腺苷二磷酸核糖聚合酶-1(PARP)、半胱氨酸天冬氨酸蛋白酶3(caspase-3)、裂解的半胱氨酸天冬氨酸蛋白酶-3(cleaved caspase-3)表达的影响。结果低、中、高剂量实验组凋亡率分别为(7.59±0.86)%,(47.60±3.67)%和(65.64±1.92)%,与对照组的(3.25±0.27)%相比,差异均有统计学意义(P<0.05或P<0.01)。对照组、低、中、高剂量实验组的Bax和Bcl-2的蛋白相对表达量比值分别为0.33±0.07,0.60±0.04,0.72±0.14和4.52±0.50;cleaved caspase-3的蛋白相对表达量分别为0.10±0.02,0.22±0.04,0.28±0.05,0.42±0.05,对照组的上述指标与低、中、高剂量实验组相比,差异均有统计学意义(P<0.05或P<0.01)。对照组、中、高剂量实验组的caspase-3的蛋白相对表达量分别为0.50±0.05,0.30±0.03,0.09±0.05,PARP的蛋白相对表达量分别为1.08±0.12,0.53±0.09,0.35±0.10,对照组上述指标与中、高剂量实验组相比,差异均有统计学意义(均P<0.01)。结论H抑制MDA-MB-231细胞的增殖,可能与其调控Bcl/Bax-2/caspase-3通路诱导细胞凋亡有关。

Objective To investigate the effect of Rhein derivative H on proliferation and apoptosis of triple negative breast cancer cell line MDA-MB-231 and its mechanism. Methods MDA-MB-231 cells were divided into control group(without H) and test-L,-M,-H group(1, 2, 4 μmol·L-1 H respectively). The changes of apoptosis were detected by flow cytometry. The expression of apoptosis pathway related proteins of B-cell lymphoma/leukemia-2(Bcl-2), Bcl-2 associated X protein(Bax), poly ADP-ribose polymerase(PARP), cysteinyl aspartate specific proteinase 3(caspase-3), cleaved cysteinyl aspartate specific proteinase 3(cleaved-caspase-3) were detected by Western blot. Results The apoptosis rates in the test-L,-M,-H group were(7.59±0.86)%,(47.60±3.67)% and(65.64±1.92)%, respectively, compared with(3.25±0.27)% in the control group the differences were significant( P < 0. 05 or P < 0. 01). The ratios of Bax/Bcl-2 in the control group,test-L,-M,-H group were 0. 33 ± 0. 07,0. 60 ± 0. 04,0. 72 ± 0. 14 and 4. 52 ± 0. 50,respectively. The protein levels of cleaved caspase-3 were 0. 10 ± 0. 02,0. 22 ± 0. 04,0. 28 ± 0. 05,0. 42 ± 0. 05,respectively. The indexes in the control group were significantly different from those in the test-L,-M,-H group( P < 0. 05 or P < 0. 01). The protein levels of caspase-3 in the control group,test-M,-H group were 0. 50 ± 0. 05,0. 30 ± 0. 03,0. 09 ± 0. 05,and the protein levels of PARP were 1. 08 ± 0. 12,0. 53 ± 0. 09,and 0. 35 ± 0. 10,respectively. Compared between control group and test-M,-H group,the differences were significant( all P < 0. 01). Conclusion H inhibits the proliferation of MDA-MB-231,and the mechanism was associated with inducing apoptosis through regulating Bcl/Bax-2/Caspase-3 pathway.