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目的研究内皮抑素对子宫内膜癌HEC1B细胞增殖、凋亡及其对聚腺苷二磷酸核糖聚合酶-1(PARP-1)和胱天蛋白酶-3(Caspase-3)表达的影响。方法用终浓度为10,50,100,200 mmol·L~(-1)的内皮抑素干预的子宫内膜癌HEC1B细胞为实验组,无任何药物处理的子宫内膜癌HEC1B细胞为对照组。培养48,72 h后,用噻唑蓝(MTT)比色法检测各组子宫内膜癌HEC1B细胞增殖情况。用Hoechst33258荧光染色法检测子宫内膜癌HEC1B细胞凋亡情况;用酶联免疫吸附(ELISA)法检测子宫内膜癌HEC1B细胞培养上清液中PARP-1和Caspase-3表达情况。结果对照组48,72 h细胞增殖抑制率分别为(6.24±0.39)%,(5.63±0.41)%,10,50,100,200 mmol·L~(-1)实验组48 h细胞增殖抑制率分别为(6.98±0.52)%,(14.36±1.02)%,(24.31±2.06)%,(28.16±2.13)%;10,50,100,200 mmol·L~(-1)实验组72 h细胞增殖抑制率分别为(8.96±0.54)%,(23.16±2.45)%,(29.48±2.81)%,(38.49±0.68)%。与对照组比较,10,50,100,200 mmol·L~(-1)实验组不同培养时间HEC1B细胞的增殖抑制率均显著升高(P<0.01),且HEC1B细胞的增殖抑制率随着培养时间的延长及药物浓度的增加而增大(P<0.05或P<0.01)。内皮抑素干预子宫内膜癌HEC1B细胞72 h后,荧光染色可见,与对照组比较,实验组细胞出现细胞核固缩、细胞核碎裂、凋亡小体形成及荧光强度增强等凋亡特征。10,50,100,200 mmol·L~(-1)实验组子宫内膜癌HEC1B细胞培养上清中PARP-1蛋白表达量低于对照组,Caspase-3蛋白表达量明显高于对照组(P<0.05或P<0.01),且呈现一定的浓度依赖性。结论内皮抑素可抑制子宫内膜癌HEC1B细胞的增殖,并促其凋亡,其机制可能与下调PARP-1表达及上调Caspase-3表达有关。
Objective To investigate the effects of endostatin on proliferation, apoptosis and the expression of PARP-1 and Caspase-3 in human endometrial carcinoma HEC1B cells. Methods HEC1B cells treated with endostatin at a final concentration of 10, 50, 100 and 200 mmol·L -1 were used as the experimental group. HEC1B cells without any drug treatment were used as the control group. After cultured for 48 and 72 h, the proliferation of HEC1 B cells in each group was detected by MTT assay. Hoechst33258 fluorescence staining was used to detect the apoptosis of HEC1B cells. The expression of PARP-1 and Caspase-3 in the culture supernatant of endometrial carcinoma HEC1B cells was detected by enzyme-linked immunosorbent assay (ELISA). Results The inhibitory rates of cell proliferation in control group at 48 and 72 h were (6.24 ± 0.39)% and (5.63 ± 0.41)%, respectively. The inhibitory rates of cell proliferation at 48 and 48 h in 10, 50, 100 and 200 mmol·L -1 groups were 6.98 (14.36 ± 1.02)%, (24.31 ± 2.06)% and (28.16 ± 2.13)%, respectively. The inhibitory rates of proliferation in the groups of 72 h after treatment with 10, 50, 100 and 200 mmol·L -1 were (8.96 ± 0.54%, 23.16 ± 2.45%, 29.48 ± 2.81%, 38.49 ± 0.68%, respectively. Compared with the control group, the proliferation inhibition rates of HEC1B cells in 10, 50, 100 and 200 mmol·L -1 experimental groups were all significantly increased (P <0.01), and the inhibition rate of HEC1B cells increased with the incubation time And drug concentration increased (P <0.05 or P <0.01). Endostatin intervention of endometrial cancer HEC1B cells 72 h after staining, compared with the control group, the experimental group of cells appeared nuclear pyknosis, nuclear fragmentation, apoptotic body formation and enhanced fluorescence intensity and other apoptosis characteristics. 10,50,100,200 mmol·L -1 experimental group, the expression of PARP-1 protein in the culture supernatant of HEC1B cells in endometrial carcinoma was lower than that in the control group, and the expression of Caspase-3 protein was significantly higher than that in the control group (P <0.05 or P <0.01), and showed a certain concentration-dependent. Conclusion Endostatin can inhibit the proliferation and promote the apoptosis of endometrial carcinoma HEC1B cells. The mechanism may be related to the downregulation of PARP-1 and the up-regulation of Caspase-3 expression.