藤梨根提取物抑制肺癌A549细胞增殖作用机制研究

目的观察藤梨根乙酸乙酯提取物（RAE）对肺癌A549细胞凋亡的影响,并探讨其作用机制。方法体外培养肺癌A549细胞,分别给予不同浓度的RAE进行干预,流式细胞术检测用药后A549细胞凋亡率变化,免疫组化SP法检侧细胞Bcl-2、Bax蛋白表达变化,RT—PCR法检测细胞Bcl-2、Bax mRNA表达,激光共聚焦显微技术测定细胞胞内Ca^2＋浓度变化。结果各治疗组给予RAE作用后细胞凋亡率明显增加,并存在时相性和量效依赖性;用药后A549细胞Bcl-2蛋白和mRNA表达降低,而Bax蛋白和mRNA表达上调,细胞内Ca^2＋浓度明显升高,与对照组比较均有显著统计学差异（P均〈0．01）,亦存在时相性和量效依赖性。结论RAE可明显诱导肺癌A549细胞凋亡,其机制可能与降低其Bcl-2蛋白和mRNA表达、上调Bax蛋白和mRNA表达、升高细胞内Ca^2＋浓度有关。

Study of the action mechanism of Radix Actinidiae extracts on proliferation of lung cancer cell A549

Objective To observe the influence of Radix Actinidiae extracts on apoptosis of A549 cell, and to study its mechanism of action. Methods Culturing A549 cell in vitro, treating with different concentrations of Radix Aetinidiae extracts, cell apoptosis was measured by FCM. Immunohistochemistry and RT-PCR was used to detect the express of Bel-2, Bax protein and mRNA. Intracellular calciumion level was measured by laser scanning confocal microscopy. Results After given the drug, apoptosis rate of A549 cell in each treat groups obviously increased by a dose-and time-dependent manner. The expressions of Bcl-2 protein and mRNA of treat groups were down-regulated and the expressions of Bax protein and mRNA of treat groups were up-regulated. Intracellular caleiumion level obviously increasing in treat groups. The change abovementioned was also in a dose and time-dependent manner. Significant difference was found compare with control group （P 〈 0. 01 ）. Conclusions Radix Actinidiae extracts can induce apoptosis of A549 cell in vitro. The mechanism of induce apoptosis may be mediated by regulating the expression of apoptosis-regulated gene and the level of intracellular calciumion level.