黄芪建中汤含药血清联合顺铂对A549细胞的抑制作用及Smad3、Smad7表达的影响

Effects of Huangqi Jianzhong DecoctionCombined with Cisplatin on Inhibition of Proliferation and Expressions of Smad3 and Smad7 in Lung Cancer A549 Cells

目的:观察黄芪建中汤含药血清联合顺铂对肺癌A549细胞中Smad3、Smad7蛋白表达的影响。方法:应用MTT法分别计算5%高、中、低剂量和10%高、中、低剂量黄芪建中汤含药血清作用于人肺腺癌A549细胞株24、48、72 h的半数抑制浓度(IC50)、黄芪建中汤最佳含药血清(10%中剂量,48h)联合最佳顺铂(5μmol/mL,48 h)作用于人肺腺癌A549细胞株的增殖抑制率。免疫组织化学显色技术检测肺癌A549细胞内Smad3、Smad7蛋白的表达情况;Western Blot技术检测Smad3、Smad7蛋白表达。结果:①黄芪建中汤含药血清作用于A549细胞株48h的(IC50)为10%中剂量。黄芪建中汤含药血清联合顺铂作用于A549细胞的增殖抑制率高于两单药组和空白组(P<0.05)。②免疫组化方法检测Smad3、Smad7在A549细胞中的表达,与其它组相比在中西药联合组(HuangQi+Cisplatin)中Smad3蛋白表达棕黄色颗粒最少,Smad7蛋白表达棕黄色颗粒最多。差异具有统计学意义(P<0.05)。③Western Blot技术检测Smad3与Smad7的表达。与其它组相比,中西药联合组Smad3表达下调,Smad7表达上调(P<0.05)。结论:黄芪建中汤含药血清联合顺铂对肺癌A549细胞具有抑制增殖作用,其作用机制可能与上调smad7蛋白表达,下调Smad3蛋白表达有关。

Objecttive:To observe the effect of HuangqiJianzhong Decoction combined with cisplatin on the expressions of Smad3 and Smad7 proteins in lung cancer A549 cells. Methods:By MTT method,we calculated half inhibitory concentration(IC50) of the drug serum of 5% and 10% HuangqiJianzhong Decoctionon human lung adenocarcinoma A549 cell line for 24, 48, 72 h and the proliferation inhibition rate of serum-containing HuangqiJianzhong Decoction(10%,middle dose, 48 h) and cisplatin(5 μmol/mL, 48 h) on human lung adenocarcinoma A549 cell line. The expression and localization of Smad3 and Smad7 proteins in lung cancer A549 cells were detected by immunohistochemical coloration. Protein expressions of Smad3 and Smad7 were detected by Western Blot. Results: The serum of Huangqi Jianzhong Decoctionon A549 cell linefor 48 h was 10% medium dose. The proliferation inhibition rate of Huangqi Jianzhong Decoction combined with cisplatin on A549 cells was higher than that of the two monotherapy groups and the blank group(P<0.05). The expressions of Smad3 and Smad7 in A549 cells were detected by immunohistochemistry. Compared with other groups, the brown-yellow particles were leastly expressed by Smad3 protein in the combination of Chinese and western medicine(Huangqi+Cisplatin), and the brown-yellow particles were mostly expressed by Smad7 protein. The difference was statistically significant(P<0.05). The expressions of Smad3 and Smad7 were detected by Western Blot. Compared with other groups, Smad3 expression was down-regulated and Smad7 expression was up-regulated(P<0.05). Conclusion: Huangqi Jianzhong Decoction combined with cisplatin can inhibit the proliferation of lung cancer A549 cells, and the mechanism may be related to up-regulation of smad7 protein expression and down-regulation of Smad3 protein expression.