LncRNA TPT1-AS1在前列腺癌中的表达及对生物学功能的影响  

Expression of LncRNA TPT1-AS1 in Prostate Cancer and Its Effect on Biological Function

作　　者：王伟[1] 温英武[1] 高双友[1] 李双利[1] 宋蕊[2] 杨康宁[1] Wang Wei;Wen Yingwu;Gao Shuangyou;Li Shuangli;Song Rui;Yang Kangning(Department of Urology Surgery,Kailuan General Hospital,Hebei Tangshan 063000,China;Department of Gastroenterology,Kailuan General Hospital,Hebei Tangshan 063000,China)

机构地区：[1]开滦总医院泌尿外科,河北唐山063000 [2]开滦总医院消化内科,河北唐山063000

出　　处：《中国药师》2020年第5期793-797,803,共6页China Pharmacist

基　　金：河北省医学科学研究课题计划项目(编号:20191336)。

摘　　要：目的:研究长链非编码RNA(LncRNA)肿瘤蛋白翻译调节因子1(TPT1)-反义RNA1(AS1)对前列腺癌细胞增殖、侵袭的影响。方法:体外培养人正常前列腺上皮细胞(RWPE-1)和前列腺癌细胞系雄激素非依赖f生前列腺癌(LNCaP)、DU-145、PC-3、22RV1,实时荧光定量PCR(qRT-PCR)检测各细胞系中LncRNA TPT1-AS1表达水平;选取表达差异最大的细胞系PC-3、DU-145,阳离子脂质体LipofectamineTM 2000瞬时转染si-RNATPT1-AS1阴性对照(si-RNA对照)组和siRNA-TPT1-AS1干扰(siRNA-TPT1-AS1干扰)组,不添加任何试剂设为空白对照组。qRT-PCR验证siRNA-TPT1-AS1转染情况;CCK-8检测TPT1-AS1-sh对细胞增殖的影响;Transwell检验TPT1-AS1-sh对细胞侵袭的影响;免疫印记(WB)法检测细胞TPT1蛋白的变化。结果:与正常前列腺细胞系RWPE-1相比,前列腺癌细胞系LNCaP、DU-145、PC-3、22RV1中TPT1-AS1的水平显著升高(P<0.05)。在DU-145、PC-3中,空白对照组与si-RNA对照组中TPT1-AS1表达量、0,6,12,24,36,48 h各时期450 nm处的吸光度(A450)值、侵袭细胞数量、TPT1蛋白表达量差异均无统计学意义(P>0.05);与空白对照组、si-RNA对照组相比,siRNATPT1-AS1干扰组中TPT1-AS1表达量,24,36,48 h细胞A450值、细胞侵袭数量、TPT1蛋白表达量显著降低(P<0.05)。在DU-145中,与空白对照组、si-RNA对照组相比,siRNA-TPT1-AS1干扰组12 h细胞A450值显著降低(P<0.05)。结论:前列腺癌细胞系中TPT1-AS1表达上调;干扰前列腺癌细胞系DU-145、PC-3中TPT1-AS1可抑制细胞增殖、侵袭。Objective:To study the effects of TPT1-AS1 on the proliferation and invasion of prostate cancer cells.Methods:Normal prostatic cell lines RWPE-1 and prostate cancer cell lines LNCaP,DU-145,PC-3 and 22RV1 were cultured in vitro,and the expression of LncRNA TPT1-AS1 in each cell line was detected by real-time fluorescence quantitative PCR(qRT-PCR);the most differentially expressed cell lines PC-3,DU-145 and cationic liposome were transfected instantaneously by LipofectamineTM 2000 used as si-RNA TPT1-AS1 negative control group and siRNA-TPT1-AS1 interference group(siRNA-TPT1-AS1 Group),and the cells without any reagent were set as the blank control group.The transfection of siRNA-TPT1-AS1 was verified by qRT-PCR,and CCK-8 was used to detect the effect of TPT1-AS1-sh on the cell proliferation,the effect of TPT1-AS1-sh on the cell invasion was examined by Transwell assay,and the change of TPT1 protein was detected by immunoblotting(WB).Results:Compared with that in the normal prostate cell line RWPE-1,the level of TPT1-AS 1 in prostate cancer cell lines LNCaP,DU-145,PC-3 and 22RV1 significantly increased(P<0.05).In DU-145 and PC-3,there were no significant differences in TPT1-AS1 expression,A450 at 0,6,12,24,36 and 48 hours,number of invasive cells and TPT1 protein expression between the blank control group and si-RNA NC group(P>0.05);compared with those of the blank control group and si-RNA NC group,the expression of TPT1-AS1,A450 at 24,36 and 48 hours,number of cell invasion and expression of TPT1 protein in siRNA-TPT1-AS1 group significantly decreased(P<0.05).In DU-145,compared with that of the blank control group and si-RNA NC group,A450 at 12 h in siRNA-TPT1-AS1 group significantly decreased(P<0.05).Conclusion:The expression of TPT1-AS1 in prostate cancer cell line is up-regulated in prostate cancer cell lines;and interfering with TPT1-AS1 in prostate cancer cell lines DU-145 and PC-3 can inhibit the cell proliferation and invasion.

关 键 词：反义长链非编码RNA肿瘤蛋白翻译调节因子1 前列腺癌 细胞系 增殖 侵袭 

分 类 号：R737.25[医药卫生—肿瘤]