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目的观察重组人p53腺病毒(rAd-p53)联合苦参碱对肺癌A549细胞生长的协同抑制作用,探讨协同抑制作用的机制。方法通过MTT及流式细胞技术观察rAd-p53和苦参碱单药与联合用药对肺癌A549细胞生长抑制率及凋亡率,用Western blot检测p53、bcl-2、bax蛋白的表达情况。结果 (1)rAd-p53、苦参碱对肺癌A549细胞均具有生长抑制作用,生长抑制率呈现时间和剂量依赖性(P<0.05)。(2)rAd-p53联合苦参碱对肺癌A549细胞生长有协同抑制作用,与单药相比,抑制率及细胞凋亡率明显增强(P<0.05);不同的联合加药顺序对细胞生长抑制没有明显差异(P>0.05)。(3)单药rAd-p53组、联合用药组的肿瘤细胞有明显的p53蛋白表达,bax蛋白在联合用药组中表达量最高,bcl-2联合用药组中表达量最低。结论重组人p53腺病毒联合苦参碱对肺癌A549细胞生长具有协同抑制作用,能抑制细胞增殖和促进细胞凋亡,且呈现时间和剂量依赖性;协同抑制作用可能通过上调促凋亡蛋白bax的表达,下调抑凋亡蛋白bcl-2的表达实现的。
Objective To observe the synergistic inhibitory effect of recombinant human p53 adenovirus (rAd-p53) and matrine on lung cancer A549 cells and to explore the mechanism of synergistic inhibition. Methods MTT and flow cytometry were used to observe the growth inhibition rate and apoptosis rate of lung cancer A549 cells treated with rAd-p53 and matrine alone. Western blot was used to detect the expression of p53, bcl-2 and bax protein. Results (1) The growth inhibitory effect of rAd-p53 and matrine on lung cancer A549 cells both in a time-and dose-dependent manner (P <0.05). (2) The rAd-p53 combined with matrine had a synergistic inhibitory effect on the growth of A549 cells. Compared with single drug, the inhibitory rate and apoptosis rate of rAd-p53 combined with matrine were significantly increased (P <0.05) There was no significant difference in inhibition (P> 0.05). (3) The single cell rAd-p53 group showed significant p53 protein expression in the tumor cells. The highest expression of bax protein was found in the combination group and the lowest in the bcl-2 combination group. Conclusions Recombinant human p53 adenovirus combined with matrine has a synergistic inhibitory effect on the growth of lung cancer A549 cells, which can inhibit cell proliferation and promote apoptosis in a dose- and time-dependent manner. The synergistic inhibitory effect may be related to upregulation of pro-apoptotic protein bax Expression and down-regulation of the expression of anti-apoptotic protein bcl-2.