食管癌细胞中Survivin对Smad3及PD-L1表达调控的影响  

Effects of Survivin on Smad3 and PD-L1 expression in esophageal cancer cells

作　　者：胡慧玲 李惠武 朱世茂 杨银银 海妮萨依姆·图尔荪 李卉 HU Hui-ling;LI Hui-wu;ZHU Shi-mao;YANG Yin-yin;HAINISAYIMU·Tuerxun;LI Hui(Department of Biochemistry and Molecular Biology,Basic Medical College,Xinjiang Medical University,Urumqi Xinjiang 830011,China;Central Laboratory,Xinjiang Medical University,Urumqi Xinjiang 830011,China;Medical Research Center,Yuebei People Hospital,Shaoguan Guangdong,512025,China)

机构地区：[1]新疆医科大学基础医学院生物化学与分子生物学教研室,新疆乌鲁木齐830011 [2]新疆医科大学中心实验室,新疆乌鲁木齐830011 [3]粤北人民医院医学研究中心,广东韶关512025

出　　处：《职业与健康》2022年第8期1040-1045,共6页Occupation and Health

基　　金：国家自然科学基金项目(81660459)。

摘　　要：目的探讨食管鳞癌Eca109细胞中,经炎性因子白介素6(interleukin-6,IL-6)诱导激活Jak2/Stat3信号通路促使程序性细胞死亡配体1(programmed cell death ligand1,PD-L1)表达升高的调控过程中,生存素(Survivin)对Smad3及PD-L1的调控作用。方法选用IL-6诱导食管鳞癌Eca109细胞6、12、24 h,激活Jak2/Stat3信号通路,再使用Jak2/Stat3信号通路抑制剂AG490干预24 h,应用免疫印迹试验(Western blot)检测p-Jak2、Jak2、p-Stat3、Stat3、Survivin、p-Smad3、Smad3和PD-L1蛋白表达量的变化;在IL-6诱导下,使用Survivin抑制剂YM155干预Eca109细胞24 h,应用Western blot方法和流式细胞术分别检测Survivin、p-Smad3、Smad3和PD-L1蛋白表达量的变化以及不同分组细胞凋亡的情况。结果使用IL-6诱导Eca109细胞6、12、24 h后,Western blot检测结果显示,p-Jak2、Jak2、p-Stat3、Stat3、Survivin、p-Smad3、Smad3和PD-L1的蛋白表达量升高,以12、24 h最为明显(均P<0.05);使用抑制剂AG490干预24 h后,p-Jak2、Jak2、p-Stat3、Stat3、Survivin、pSmad3、Smad3和PD-L1的蛋白表达水平均下调(均P<0.01);抑制剂YM155在抑制Survivin表达的同时降低了p-Smad3、Smad3和PD-L1的表达(均P<0.05);使用YM155干预后,促进食管鳞癌Eca109细胞的凋亡(P<0.05)。结论在食管鳞癌Eca109细胞中,IL-6激活Jak2/Stat3信号通路后,存在Survivin对Smad3和PD-L1的正向调控关系。提示Survivin可能通过参与调节肿瘤的免疫逃逸,从而影响食管癌的发生发展,为靶向PD-L1的免疫治疗提供可靠的实验依据以及理论基础。Objective To explore the regulation of Survivin on Smad3 and PD-L1 in esophageal squamous carcinoma Eca109 cells induced by interleukin-6(IL-6)and activated Jak2/Stat3 signaling pathway to promote programmed cell death ligand1(PD-L1)expression.Methods IL-6 was selected to induce Eca109 cells of esophageal squamous cell carcinoma for 6,12 and 24 hours,and Jak2/Stat3 signaling pathway was activated,and then AG490,an inhibitor of Jak2/Stat3 signaling pathway,was used to intervene for 24 hours.The protein expression levels of p-Jak2,Jak2,p-Stat3,Stat3,Survivin,p-Smad3,Smad3 and PD-L1 were detected by Western blot.Under the induction of IL-6,Survivin inhibitor YM155 was used to interfere Eca109 cells for 24 hours.The Western blot and flow cytometry were used to detect the protein expression of Survivin,p-Smad3,Smad3 and PD-L1,and the apoptosis of cells in different groups,respectively.Results Eca109 cells were induced by IL-6 for 6,12 and 24 hours,Western blot analysis showed that the protein expression levels of p-Jak2,Jak2,p-Stat3,Stat3,Survivin,p-Smad3,Smad3 and PD-L1 were increased,and the changes of 12 and 24 hours were the most significant(both P<0.05).After treatment with AG490 for 24 hours,the protein expression levels of p-Jak2,Jak2,p-Stat3,Stat3,Survivin,p-Smad3,Smad3 and PD-L1 were all down-regulated(all P<0.01).The inhibitor YM155 inhibited the expression of Survivin while decreased the expression of p-Smad3,Smad3 and PD-L1(all P<0.05).After YM155 intervention,the apoptosis of esophageal squamous cell carcinoma Eca109 cells was promoted(P<0.05).Conclusions In esophageal squamous cell carcinoma Eca109 cells,Survivin positively regulates Smad3 and PD-L1 after IL-6activates the Jak2/Stat3 signaling pathway.It is suggested that Survivin may affect the occurrence and development of esophageal cancer by regulating the immune escape of tumor,which provides reliable experimental basis and theoretical basis for the immunotherapy targeting PD-L1.

关 键 词：ECA109细胞 JAK2/STAT3信号通路 生存素 SMAD3蛋白 程序性细胞死亡配体1 

分 类 号：R393[医药卫生—基础医学]