天花粉蛋白对人宫颈癌细胞增殖及凋亡的影响及其机制研究

Effect of trichosanthin on proliferation and apoptosis of human cervical cancer cells and research on its mechanism

目的研究天花粉蛋白(TCS)对人宫颈癌细胞(HeLa细胞)诱导凋亡的作用及机制。方法低剂量(7 μmol/L)、中剂量(14 μmol/L)和高剂量(28 μmol/L)TCS处理HeLa细胞10~82 h后,观察细胞形态及细胞生长情况。同样梯度剂量的TCS处理HeLa细胞24 h后,检测细胞周期,凋亡变化,应用蛋白免疫印迹法检测细胞凋亡相关蛋白表达。结果 TSC处理后HeLa细胞形态皱缩,贴壁变差,增殖减少,呈剂量依赖性,低、中剂量TCS组随时间延长,细胞数逐步减少(r =-0.9727、-0.9757,P < 0.05)。处理24 h后中、高剂量TCS组G0/G1期比例较空白对照组增加(P < 0.05),低、中、高剂量TCS组S期比例较空白对照组降低,差异有统计学意义(P < 0.05)。低、中、高剂量TCS组HeLa细胞凋亡率较空白对照组增加(P < 0.05)。与空白对照组比较,低、中、高剂量TCS组Caspase-3、Caspase-9、聚腺苷二磷酸核糖聚合酶(PARP)和磷酸化细胞外调节蛋白激酶(p-ERK)蛋白表达水平逐渐减少,与TCS呈现浓度依赖关系(r =-0.969,P < 0.05)。结论 TCS通过活化Caspase家族蛋白、诱导细胞凋亡,进而抑制宫颈癌HeLa细胞增殖,从而发挥抗肿瘤作用。

Objective To study the effect and mechanism of trichosanthin (TCS) on inducing apoptosis of human cervical cancer HeLa cells. Methods HeLa cells were treated with low (7 μmol/L), medium (14 μmol/L) and high (28 μmol/L) doses of TCS for 10-82 h, the cell morphology and inhibition of cell growth was observed. The cell cycle and apoptosis was detected and Western blot was performed to detect the protein expression of apoptosis proteins of cervical cancer cells HeLa after treated with TCS for 24 h. Results HeLa cells of cervical cancer treated by TSC showed wrinkles and deformation, the growth of HeLa cells was inhibited. The number of cells in low and medium dose of TCS group decreased with time (r =-0.9727,-0.9757, P < 0.05). Compared with the blank control group, the G0/G1 phase of HeLa cells in medium, high dose of TCS group were significantly increased (P < 0.05), and the S phase in low, medium, high dose of TCS group were decreased after TCS treated for 24 h, the differences were statistically significant (P < 0.05). The apoptosis rate of HeLa cells in the low, medium and high dose of TCS group was increased compared with that in the blank control group (P < 0.05). Compared with the blank control group, the protein expressions levels of Caspase-9, Caspase-3, polyadenosine diphosphate ribosome polymerase (PARP) and p-extracellular regulated protein kinase (ERK) in low, medium, high dose of TCS group were gradually decreased in a dose-dependent manner after of TCS treatment (r =-0.969, P < 0.05). Conclusion TCS can induce apoptosis by activating the Caspase family of proteins to inhibit the proliferation of cervical cancer HeLa cells to exert anti-tumor effects.