IL-6调控STAT3/Notch信号通路促进胰腺癌细胞增殖 迁移及化疗抵抗的机制研究

Mechanisms of IL-6 regulating STAT3/Notch signaling pathway to promote the proliferation,migration and chemotherapy resistance of pancreatic cancer cells

目的观察白细胞介素(IL-6)对胰腺癌细胞的影响,并分析其机制。方法选取对数生长期胰腺癌细胞,随机分组:对照组,正常培养;IL-6组:IL-6终浓度为50 ng·ml^(-1)的培养基培养;IL-6+JSI-124组:含50 ng·ml^(-1) IL-6和0.1μmol·L^(-1) JSI-124(STAT3磷酸化特异性抑制剂)的培养基培养;IL-6+DAPT组:含50 ng·ml^(-1) IL-6和50μmol·L^(-1) DAPT(Notch通路抑制剂)的培养基培养。Western blot检测p-STAT3/STAT3、Notch1、Hes1蛋白表达,transwell实验检查细胞迁移。建立顺铂耐药细胞模型,分析Ki-67、MMP-9和P-gp蛋白水平。裸鼠成瘤实验验证IL-6对瘤体STAT3/Notch信号通路的影响。结果与IL-6组比较,IL-6+JSI-124组和IL-6+DAPT组p-STAT3/STAT3、Notch1、Hes1表达水平下调,增值率降低,迁移水平下调(P<0.05);与IL-6组比较,IL-6+JSI-124组和IL-6+DAPT组增殖率下调,IC 50降低,Ki-67、MMP-9和P-gp表达下调(P<0.05)。与顺铂组抑瘤率[(67.12±2.32)%]比较,IL-6+顺铂组抑瘤率[(21.71±1.37)%]明显下调(P<0.01),p-STAT3/STAT3、Notch1、Hes1表达上调(P<0.05)。结论IL-6调节STAT3/Notch通路促进胰腺癌sw1990细胞增殖、迁移和耐药。

Objective To observe the effect of IL-6 on pancreatic cancer cells and analyze its mechanism.Methods Pancreatic cancer cells in logarithmic phase were randomly divided into:control group was normally cultured;IL-6 group was cultured with the final concentration of 50 ng·ml^(-1) IL-6;IL-6+JSI-124 group was cultured with 50 ng·ml^(-1) IL-6 and 0.1μmol·L^(-1) JSI-124(STAT3 phosphorylation specific inhibitor);IL-6+DAPT group was cultured with 50 ng·ml^(-1) IL-6 and 50μmol·L^(-1) DAPT(Notch pathway inhibitor).The expression levels of p-STAT3,STAT3,Notch1 and Hes1 were detected with Western blot,cell migration with Transwell test.The cis-platin resistant cell model was established,levels of Ki-67,MMP-9 and P-gp protein were analyzed.Influences of IL-6 on tumor STAT3/Notch signaling pathway were verified via the tumor formation in nude mice.Results Compared with IL-6 group,the expression levels of p-STAT3/STAT3,Notch1 and Hes1 as well as proliferation rates and migration levels in IL-6+JSI-124 and IL-6+DAPT group lowered(P<0.05);compared with IL-6 group,proliferation rates,IC 50 and the expression levels of Ki-67,MMP-9 and P-gp in IL-6+JSI-124 and IL-6+DAPT group lowered(P<0.05).Compared with the tumor inhibition rate of cisplatin group[(67.12±2.32)%],that of cisplatin+IL-6 group[(21.71±1.37)%]significantly lowered(P<0.01),and the expression levels of p-STAT3/STAT3,Notch1 and Hes1 significantly increased(P<0.05).Conclusion IL-6 regulating STAT3/Notch signaling pathway promotes the proliferation,migration and drug resistance of pancreatic cancer sw1990 cells.