葛根粗提物、葛根素对肺癌H446细胞增殖的抑制作用及其机制

Effect and mechanism of pueraria crude extract、pure puerarin on Lung Cancer H446 Cell proliferation

目的观察葛根粗提物(CP)和葛根素纯品(SP)对小细胞肺癌H446细胞增殖的影响,并探讨其机制。方法用含不同浓度的SP和CP培养H446细胞,用MTT法检测细胞生长抑制率。用流式细胞术检测细胞周期和凋亡率、细胞免疫化学技术检测Bcl-2和Bax蛋白。另设空白对照组和溶剂对照组。结果CP作用72h的半数抑制浓度(IC50)为435μg/ml,SP为1403μg/ml。此浓度作用72h,细胞凋亡率分别为14.71%和2.61%,两组相比P<0.05;G0/G1期细胞构成比分别为79.20%和72.20%,G2/M期细胞构成比分别为8.94%和10.50%,与对照组相比P均<0.05;CP作用于H446细胞后,细胞内Bcl-2蛋白表达量降低,Bax的蛋白表达量增加,与对照组相比P均<0.05。结论SP和CP对小细胞肺癌H446细胞具有增殖抑制作用,呈浓度相关性;SP和CP可诱导细胞凋亡且CP强于SP,其机制可能与阻滞细胞周期于G0/G1期、上调Bax表达、下调Bcl-2表达有关。

Objective To observe the effect of pure puerarin and pueraria crude extract on H446 cell proliferation, and explore its mechanism. Methods H446 cells grew in different concentrations of SP and CP, cell growth inhibition was detected by MTT assay. Cell cycle and apoptosis were detected by flow cytometry. Bcl-2 and Bax protein were detected by immunohistochemistry. Results The CP for 72 h 50% inhibitory concentration （IC50） was 435 μg/ml, for SP was 1 403 μg/ml, 435μg/ml on H446 for 72 h,the apoptosis rate induced by CP and SP were respectively 14. 71% and 2. 61%, the percentages of G0/G1 phase cell numbers were respectively 79.20% and 72.20%, those of G2/M phase were respectively 8. 94% and 10. 50%, there was significant difference compared with control group（P 〈 0. 05）. Compared with the control group the Bcl-2 protein expression was lower, Bax expression is increased, P 〈 0. 05. The expression proportion of Bax/ Bcl-2 was higher than control group. Concolusions SP and CP inhibit H446 tumor cell proliferation. SP and CP can induce apoptosis, which may be related to block the cell cycle in the G0/G1 phase, up Bax Expression, lower Bcl-2 expression.