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目的评价表没食子儿茶素没食子酸酯(EGCG)联合盐酸吉西他滨对胰腺癌PANC-1细胞凋亡及对蛋白酶活化受体2(PAR-2)表达的影响。方法培养PANC-1细胞,分为不添加任何药物的对照组、60μg/mL EGCG处理组、20μg/mL吉西他滨处理组、60μg/mL EGCG联合20μg/mL吉西他滨处理组。Annexin V-FITC/PI联合染色结合流式细胞术检测各组PANC-1细胞凋亡情况,并采用实时荧光定量PCR(qRT-PCR)检测各组PANC-1细胞PAR-2 mRNA的表达。Western blot法检测各组PANC-1细胞PAR-2蛋白的表达。结果联合处理组对PANC-1细胞凋亡的诱导作用最强,依次为EGCG处理组、吉西他滨处理组。各组与对照组相比,差异均有统计学意义(P<0.01)。qRT-PCR和Western blot法结果表明联合处理组的PAR-2 mRNA和蛋白的相对表达量低于对照组(P<0.01或P<0.05)。结论 EGCG可以增强盐酸吉西他滨对胰腺癌PANC-1细胞凋亡的诱导作用,该作用与下调细胞PAR-2表达有关。
Objective To evaluate the effect of epigallocatechin-3-gallate (EGCG) combined with gemcitabine hydrochloride on the apoptosis of pancreatic cancer PANC-1 cells and the expression of protease-activated receptor 2 (PAR-2). Methods PANC-1 cells were cultured and divided into control group without any drug, EGCG 60μg / mL, gemcitabine 20μg / mL, EGCG 60μg / mL and gemcitabine 20μg / mL. The apoptosis of PANC-1 cells in each group was detected by Annexin V-FITC / PI combined with flow cytometry. The expression of PAR-2 mRNA in PANC-1 cells was detected by real-time quantitative PCR (qRT- Western blot was used to detect the expression of PAR-2 protein in PANC-1 cells. Results The combined treatment group had the strongest induction effect on apoptosis of PANC-1 cells, followed by EGCG treatment group and gemcitabine treatment group. Compared with the control group, the difference was statistically significant (P <0.01). The results of qRT-PCR and Western blot showed that the relative expression of PAR-2 mRNA and protein in the combined treatment group was lower than that in the control group (P <0.01 or P <0.05). Conclusion EGCG can enhance the induction of apoptosis of pancreatic cancer PANC-1 cells by gemcitabine hydrochloride, which may be related to the down-regulation of PAR-2 expression.