ATF6通过Hippo-YAP信号通路促进前列腺癌PC3细胞的增殖、迁移和侵袭  

Activating transcription factor 6 promotes proliferation, migration and invasion of PC3 human prostate cancer cells through Hippo-YAP signaling pathway

作　　者：刘恒川 杨宗珂 刘清源 张金栋 朱辉轩 王德林[1] LIU Hengchuan;YANG Zongke;LIU Qingyuan;ZHANG Jindong;ZHU Huixuan;WANG Delin(Department of Urology,the First Affiliated Hospital of Chongqing Medical University,Chongqing,400016;Department of Urology,Dianjiang People's Hospital of Chongqing,Chongqing,408399,China)

机构地区：[1]重庆医科大学附属第一医院泌尿外科,重庆400016 [2]重庆市垫江县人民医院泌尿外科,重庆408399

出　　处：《陆军军医大学学报》2023年第1期29-37,共9页Journal of Army Medical University

基　　金：重庆市自然科学基金(cstc2019jcyj-msxmX0732);重庆市社会事业与民生保障科技创新专项(cstc2017shms-zdyf0319)。

摘　　要：目的 探讨转录活化因子6 (activating transcription factor 6, ATF6)对前列腺癌PC3细胞增殖、迁移和侵袭的影响。方法 收集前列腺癌组织标本和前列腺增生组织标本,实时荧光定量PCR(qRT-PCR)和Western blot分别检测ATF6的mRNA和蛋白质水平。通过shRNA慢病毒干扰载体感染PC3细胞,敲降ATF6表达水平。CCK8和EdU实验检测PC3细胞增殖能力,划痕实验和Transwell侵袭实验检测PC3细胞迁移和侵袭能力。采用裸鼠异种移植模型评估ATF6在体内对肿瘤增殖的影响。使用JASPAR数据库预测和荧光素酶报告基因评估Yes相关蛋白(YAP)和ATF6之间的关联。qRT-PCR、免疫荧光检测YAP的mRNA和蛋白质水平。qRT-PCR检测YAP的两个下游因子结缔组织生长因子(CTGF)和富含半胱氨酸的61(Cyr61)的mRNA表达水平。结果 在前列腺癌组织中,ATF6的mRNA和蛋白质水平均显著升高(P<0.05)。敲降ATF6后,PC3细胞的增殖能力、侵袭能力和迁移能力均受到抑制(P<0.05)。动物实验证实敲降ATF6能明显抑制肿瘤生长(P<0.05)。JASPAR数据库预测发现ATF6与YAP之前存在启动结合位点,荧光素酶报告基因证实ATF6与YAP存在相互作用。敲降ATF6后显著下调YAP的mRNA及蛋白质表达水平(P<0.05),且YAP下游基因CTGF和Cyr61的mRNA表达水平也随之下调(P<0.05)。结论 ATF6能通过Hippo-YAP信号通路促进前列腺癌PC3细胞的增殖、迁移和侵袭。Objective To investigate the effects of activating transcription factor 6(ATF6) on the proliferation, migration and invasion of prostate cancer PC3 cells. Methods Prostate cancer tissue samples and benign prostatic hyperplasia tissue samples were collected, then the mRNA and protein levels of ATF6 were detected by qRT-PCR and Western blotting, respectively. Lentivirus with ATF6 shRNA was transfected into PC3 cells to downregulate ATF6 expression, then CCK8 and EdU assays were adopted to determine the proliferation of PC3 cells, and wound-healing and Transwell invasion tests were performed to evaluate the migration and invasion abilities of PC3 cells. Nude mouse xenograft models were used to assess the influence of ATF6 on tumorigenesis in vivo. Moreover, the association between Yes-associated protein(YAP) and ATF6 was analyzed using online JASPAR database prediction and luciferase reporter. The mRNA and protein levels of YAP were detected with qRT-PCR and immunofluorescence assay, and the mRNA levels of 2 downstream factors of YAP, connective tissue growth factor(CTGF) and cysteine-rich 61(Cyr61), were also measured by qRT-PCR. Results The mRNA and protein levels of ATF6 in prostate cancer tissues were significantly increased(P<0.05). Knockdown of ATF6 decreased the proliferation, invasion, and migration abilities of PC3 cells(P<0.05), and the in vivo study showed that ATF6-knockdown attenuated tumor growth as well(P<0.05). JASPAR database predicted that there was an initiation binding site between ATF6 and YAP, and the luciferase reporter assay also confirmed the interaction between them. In addition, YAP expression was remarkably downregulated after knockdown of ATF6(P<0.05), and the mRNA level of YAP downstream genes CTGF and Cyr61 were also decreased accordingly(P<0.05). Conclusion ATF6 promotes the proliferation, migration, and invasion of prostate cancer PC3 cells through Hippo-YAP signaling pathway.

关 键 词：前列腺癌 转录活化因子6 HIPPO YAP 增殖 

分 类 号：R394.3[医药卫生—医学遗传学] R73-37[医药卫生—基础医学]