miR-576靶向ALK4调控JAK/STAT通路传导对前列腺癌细胞增殖和迁移的影响  

Up-regulated expression of miR-576 inhibits ALK4 expression,regulates JAK/STAT signaling pathway and promotes proliferation and migration of prostatic cancer cells

作　　者：张一川[1] 龙智[1] 万波[1] 彭东毅[1] 魏竞超 赵雄剑 王宁[1] 高军 徐志鹏 何乐业[1] ZHANG Yi-chuan;LONG Zhi;WAN Bo;PENG Dong-yi;WEI Jing-chao;ZHAO Xiong-jian;WANG Ning;GAO Jun;XU Zhi-peng;HE Le-ye(Department of Urology,The Third Xiangya Hospital,Central South University,Changsha,Hunan 410013,China)

机构地区：[1]中南大学湘雅三医院泌尿外科,湖南长沙410013

出　　处：《中华男科学杂志》2022年第2期99-106,共8页National Journal of Andrology

基　　金：湖南省自然科学基金青年基金(2018JJ3702);新湘雅人才工程-精医助跑计划(JY201523)。

摘　　要：目的:前列腺癌进展的分子机制目前尚不明确,本研究拟探讨miR-576/ALK4轴在前列腺癌进展过程中的作用机制。方法:生物信息学分析得到可能调控ALK4表达并影响前列腺癌进展的miR-576,通过miR-576 mimics/inhibitor转染前列腺癌细胞,MTT实验检测细胞增殖,细胞划痕实验检测细胞迁移距离,双荧光素酶报告实验验证miR-576和ALK4的靶向调控关系,qPCR检测miR-576对ALK4及JAK/STAT信号通路因子JAK2、STAT3 mRNA水平的影响,Western印迹检测ALK4及JAK/STAT信号通路因子JAK2,STAT3和JAK2,STAT3磷酸化水平的变化,sh-ALK4和JAK/STAT信号通路抑制剂Ruxolitinib共同处理C4-2细胞,检测细胞增殖和迁移变化。结果:生物信息学分析和结合位点预测得到前列腺癌中表达上调并可能靶向ALK4的miR-576,双荧光素酶报告实验表明miR-576能够靶向调控ALK4,同时能够影响JAK2、STAT3磷酸化水平,功能实验结果显示miR-576过表达能够促进前列腺癌细胞增殖和迁移,而敲低miR-576则能够抑制前列腺癌细胞增殖和迁移,回复实验结果显示sh-ALK4能够促进细胞增殖和迁移,而抑制剂Ruxolitinib能够抑制sh-ALK4的促增殖和迁移作用。结论:miR-576在前列腺癌中表达上调,并可抑制ALK4的表达,调控JAK/STAT信号通路活性,促进前列腺癌细胞增殖和迁移。Objective:To explore the mechanism of the action of the miR-576/ALK4 axis on the progression of prostate cancer(PCa).Methods:PCa cells were transfected with miR-576 mimics/inhibitor,the proliferation and migration distance of the cells were detected by MTT and scratch wound healing assay,respectively.The targeted regulation effect of miR-576 on ALK4 was verified by dual-luciferase reporter assay.The effects of miR-576 on the mRNA and protein expressions and phosphorylation levels of the ALK4 and JAK/STAT signaling pathway factors JAK2 and STAT3 were determined by qPCR and Western blot,respectively.The C4-2 cells were co-treated with sh-ALK4 and Ruxolitinib for measurement of the proliferation and migration of the PCa cells.Results:Bioinformatics analysis and binding site prediction showed that miR-576 was up-regulated in the PCa cells,and dual-luciferase reporter assay revealed its targeted regulation effect on ALK4 and its impact on the phosphorylation levels of JAK2 and STAT3.Overexpressed miR-576 promoted while knocked-down miR-576 inhibited the proliferation and migration of the PCa cells.sh-ALK4 increased the proliferation and migration of the cells,while Ruxolitinib suppressed the promoting effect of sh-ALK4.Conclusion:The expression of miR-576 is up-regulated in PCa,inhibits the expression of ALK4,regulates the activity of the JAK and STAT signaling pathways,and promotes the proliferation and migration of PCa cells.

关 键 词：前列腺癌 miR-576 间变性淋巴瘤激酶4 JAK/STAT信号通路 

分 类 号：R737.25[医药卫生—肿瘤]