摘要
目的:探讨lncRNA FLJ26245在前列腺癌组织和细胞中的表达及其对PC-3细胞增殖与迁移能力的影响及其分子机制。方法:选用2017年3月至2019年5月在洛阳中心医院手术切除的52例前列腺癌及相应的癌旁组织标本,以及前列腺细胞系LNCaP、C4-2B、PC-3、DU-145和正常前列腺上皮细胞RWPE-1,用qPCR法检测前列腺癌组织和细胞中FLJ26245的表达水平。分别将FLJ26245 mimic和阴性对照质粒(lncR-NC)转染到PC-3细胞中,用MTT法、细胞划痕愈合实验分别检测细胞的增殖和迁移能力。生物信息学技术预测和双荧光素酶基因报告实验验证FLJ26245与miR-200a-3p、酪氨酸磷酸酶受体G(PTPRG)三者之间的靶向关系。用qPCR和WB法分别检测FLJ26245下游基因及增殖与迁移相关蛋白的表达。结果:FLJ26245在前列腺癌组织和细胞中的表达水平分别显著低于癌旁组织(P<0.01)和RWPE-1细胞(P<0.01或P<0.05),以PC-3细胞中的表达水平为最低(P<0.01)。FLJ26245过表达可明显抑制PC-3细胞的增殖和迁移能力(P<0.05或P<0.01)。FLJ26245可与miR-200a-3p靶向结合,miR-200a-3p可与PTPRG靶向结合(均P<0.01)。FLJ26245过表达的PC-3细胞中miR-200a-3p表达水平显著降低(P<0.01)、PTPRG mRNA和蛋白表达水平均明显升高(均P<0.01),细胞中增殖和迁移相关蛋白cyclin A、CDK2和Twist、N-cadherin均显著降低(均P<0.01)。结论:FLJ26245在前列腺癌组织及细胞中均低表达,其可能通过miR-200a-3p/PTPRG轴调控前列腺癌PC-3细胞的增殖与迁移能力。
Objective:To observe the expression level of long non-coding RNA FLJ26245(lncRNA FLJ26245)in prostate cancer tissues and cells,and to explore its effect on the proliferation and migration of prostate cancer PC-3 cells and the underlying mechanism.Methods:A total of 52 pairs of prostate cancer tissues and corresponding para-cancerous tissues from patients who underwent surgery at Luoyang Central Hospital from March 2017 to May 2019 were collected for this study;in addtion,the prostate cancer cell lines(LNCaP,C4-2 B,PC-3,DU-145)and normal prostate epithelial cells(RWPE-1)were also selected.qPCR was used to detect the expression level of FLJ26245 in prostate cancer tissues and cells.FLJ26245 mimics and negative control plasmids(lncR-NC)were transfected into PC-3 cells,respectively.MTT assay and Wound-healing test were used to detect the proliferation and migration ability of prostate cancer cells.Bioinformatics technology was used to predict the targeting relationship among FLJ26245,miR-200 a-3 p,and tyrosine phosphatase receptor G(PTPRG),which was further validated with Dual luciferase reporter gene assay.qPCR and WB methods were used to detect the expression of FLJ26245 downstream gene and proliferation and migration related proteins,respectively.Results:Compared with para-cancerous tissues,the expression level of FLJ26245 in prostate cancer tissues was significantly downregulaed(P<0.01).Compared with RWPE-1 cells,the expression level of FLJ26245 in prostate cancer cells was also significantly decreased(P<0.01 or P<0.05),with the lowest expression in PC-3 cells(P<0.01).Overexpression of FLJ26245 could significantly inhibit the proliferation and migration ability of PC-3 cells(P<0.05 or P<0.01).FLJ26245 could bind to miR-200 a-3 p,and miR-200 a-3 p could bind to PTPRG(all P<0.01).After overexpression of FLJ26245,the expression of miR-200 a-3 p in PC-3 cells was significantly decreased(P<0.01),the mRNA and protein expressions of PTPRG were significantly increased(all P<0.01),and proliferation and migration related proteins,such as cyclin A,CDK2 and Twist,N-cadherin,were significantly reduced(all P<0.01).Conclusion:FLJ26245 is low expressed in prostate cancer tissues and cells.FLJ26245 may inhibit the proliferation and migration of prostate cancer PC-3 cells by regulating the miR-200 a-3 p/PTPRG axis.
作者
杨凌博
杨金辉
鲁帅奇
李小辉
卢绩
YANG Lingbo;YANG Jinhui;LU Shuaiqi;LI Xiaohui;LU Ji(Department of Urology,Luoyang Central Hospital of Zhengzhou University,Luoyang 471009,Henan,China;Department of Urology,the First Affiliated Hospital of Jilin University,Changchun 130021,Jilin,China)
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
北大核心
2021年第7期659-664,共6页
Chinese Journal of Cancer Biotherapy
基金
国家自然科学基金资助项目(No.31501052)。
