农业生物技术学报2024，Vol.32Issue(5)：1111-1120,10.DOI:10.3969/j.issn.1674-7968.2024.05.012
LncRNA-TUG1通过调节miR-31-5p/YWHAE轴影响22Rv1细胞的增殖和迁移
LncRNA-TUG1 Affects 22Rv1 Cell Proliferation and Migration by Modulating miR-31-5p/YWHAE Axis
摘要
Abstract
Tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activating protein ε(YWHAE)was highly expressed in prostate cancer,and miR-31-5p could target and regulate the expression of YWHAE to affect the proliferation of prostate cancer cells.However,it was not clear which lncRNAs were involved in the targeted regulation of miR-31-5p against YWHAE.In this study,3 LncRNAs with long non-coding RNA tauronic upregulation gene(LncRNA-TUG1),LINC02604 and PDCD6IP-DT were screened to target and regulate the expression of miR-31-5p based on the results of previous studies and bioanalysis software.The expression of 3 LncRNAs in prostate cancer LNCaP,PC3,22Rv1 and normal prostate stromal immortem cells(WPMY-1)was detected by RT-qPCR,and the optimal LncRNAs regulating the miR-31-5p/YWHAE axis were screened.The effects of interfering LncRNA-TUG1 and overexpression of miR-31-5p on the expression of LncRNA-TUG1,miR-31-5p and YWHAE in 22Rv1 cells were detected by RT-qPCR.Meanwhile,the effects of co-transfection of sh-TUG1 and miR-31-5p mimics on the proliferation and migration of 22Rv1 cells were detected by CCK-8 and cell scratch assay.Western blot was used to detect the expression of proliferating cell nuclear antigen(PCNA),B cell lymphoma-2(BCL-2),Bcl-2 associated X protein(BAX)and cysteine proteinase-3(Caspase-3).The results showed that LncRNA-TUG1,LINC02604 and PDCD6IP-DT were the 3 LncRNAs regulating the miR-31-5p/YWHAE axis,and compared with normal cells,LncRNA-TUG1 was highly expressed in all PCa cell lines,especially in 22Rv1 cells.Taking LncRNA-TUG1 as the research object,after transfecting sh-TUG1 and miR-31-5p mimics into 22Rv1 cells,respectively,qPCR results showed that the expression of miR-31-5p in the miR-31-5p mimics group was significantly up-regulated compared with the NC mimics group(P＜0.01).The expression of LncRNA-TUG1 and YWHAE genes was significantly down-regulated(P＜0.05).Compared with the sh-NC group,there was no significant change in the expression of miR-31-5p in the sh-TUG1 transfection group,and the expression of LncRNA-TUG1 and YWHAE were significantly down-regulated(P＜0.05).CCK-8 and cell scratch experiments showed that co-transfection of sh-TUG1 and miR-31-5p mimics could significantly inhibit cell proliferation and migration.In addition,Western blot results showed that co-transfection of sh-TUG1 and miR-31-5p mimics could significantly inhibit the expression of PCNA and BCL-2/BAX protein,and promoted the expression of caspase-3 protein.These results indicated that LncRNA-TUG1 could regulate the expression of PCNA,BCL-2,BAX and caspase-3 proteins by regulating miR-31-5p/YWHAE,and further affected the proliferation of 22Rv1 cells.This study provides theoretical basis and experimental ideas for the study of targeted drugs for prostate cancer.关键词
LncRNA-牛磺酸上调基因(LncRNA-TUG1)/miR-31-5p/前列腺癌/22Rv1/细胞增殖/细胞迁移Key words
LncRNA-tauronic upregulation(LncRNA-TUG1)/miR-31-5p/Prostate cancer/22Rv1/Cell proliferation/Cell migration分类
农业科技引用本文复制引用
赵佳福,李永,周明帅,温晓艳,陆情梅,刘彬..LncRNA-TUG1通过调节miR-31-5p/YWHAE轴影响22Rv1细胞的增殖和迁移[J].农业生物技术学报,2024,32(5):1111-1120,10.基金项目
贵州省科学技术基金(黔科合基础[2020]1Y085) （黔科合基础[2020]1Y085）
