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目的:探讨十全大补汤对肺腺癌顺铂耐药细胞株(A549/DDP)中Wnt/β-catenin信号通路蛋白的干预作用。方法:以SD大鼠制备高、中、低剂量(65,32.5,16.25 g·kg-1,10 m L·kg-1)的十全大补汤含药血清和空白对照血清(生理盐水10 m L·kg-1)。培养细胞分为十全大补汤含药血清高、中、低剂量组和空白血清组,采用MTT法检测各组50%抑制浓度(IC50),采用蛋白质印迹法检测各组糖原合成酶激酶-3β(GSK-3β),p-GSK-3βser9,β-连环蛋白(β-catenin)及survivin蛋白表达。结果:随着十全大补汤含药血清浓度的升高,顺铂(DDP)对A549/DDP细胞的IC50逐渐降低,以中、高剂量组最显著(P<0.01);十全大补汤含药血清对A549/DDP细胞浆GSK-3β蛋白的表达无影响,但可显著下调胞浆p-GSK-3βser9蛋白的表达及细胞总β-catenin和survivin蛋白的表达,以中、高剂量组最明显(P<0.01)。结论:十全大补汤可通过降低A549/DDP细胞浆p-GSK-3βser9蛋白及细胞总β-catenin,survivin蛋白的表达而影响Wnt/β-catenin信号转导通路,进而发挥对顺铂耐药的增敏作用。
Objective: To investigate the intervention of Shidu Dabu Tang on Wnt / β-catenin signaling pathway in A549 / DDP cell line. Methods: SD rats were prepared high, medium and low doses (65,32.5,16.25 g · kg-1, 10 m L · kg-1) of Shi Dabu Tang containing serum and blank control serum (saline 10 m L · kg-1). Cultured cells were divided into Shiquanbutang Decoction serum containing high, medium and low dose groups and blank serum group, MTT assay 50% inhibitory concentration (IC50), using Western blotting assay glycogen synthase kinase -3β (GSK-3β), p-GSK-3βser9, β-catenin and survivin protein expression. Results: With the increase of serum concentration of Shiquan Dabu Tang, the IC50 of cisplatin (DDP) on A549 / DDP cells decreased gradually, especially in medium and high dose groups (P <0.01) Serum containing Tang had no effect on the expression of GSK-3β in cytoplasm of A549 / DDP, but could significantly down-regulate the expression of cytoplasmic p-GSK-3βser9 and the expression of total β-catenin and survivin in medium and high doses Group was the most obvious (P <0.01). Conclusion: Shiquan Dabu Tang can reduce Wnt / β-catenin signal transduction pathway by decreasing the expression of p-GSK-3βser9 protein and total β-catenin, survivin in A549 / DDP cytoplasm, Sensitization of the drug.