Expression of long non-coding RNA OIP5-AS1 in renal carcinoma cells and its relationship with cell proliferation and metastasis
Objective To investigate the expression of long non-coding RNA(lncRNA)OIP5-AS1 in renal carcinoma cells and its relationship with cell proliferation and metastasis.Methods The differen-tial expression of lncRNA in human tubular epithelial cell line(HK2)and renal carcinoma cell line(ACHN)was analyzed by transcriptomics.LncRNA with the most significant expression difference was se-lected as the study object,and the expression level of lncRNA OIP5-AS1 in normal renal epithelial cells and renal cancer cell lines(ACHN,GRC-1 and 786-0)was analyzed by fluorescence quantitative polymer-ase chain reaction(PCR).Renal cancer Cell lines(ACHN)were divided into lncRNA control group and lncRNA OIP5-AS1 KD group.Lentivirus was used to establish stable transfected cell lines.Cell prolifera-tion capacity of the two groups was analyzed by cell counting kit-8(CCK-8)and tumor formation assays.Cell cycle was analyzed by flow cytometry.The migration and invasion ability of the two groups of cells was analyzed by Transwell.The expression levels of proteins related to cell proliferation,invasion and epithelial mesenchymal transformation were detected by Western blotting.T test was used to compare measurement da-ta between groups.Results The most differentially expressed gene was lncRNA OIP5-AS1.The expres-sion level of OIP5-AS1 in human renal tubular epithelial cell line(HK2)was significantly lower(0.97±0.13)than that in renal carcinoma cell lines(ACHN,GRC-1 and 786-0)(2.12±0.16,1.68±0.12,1.46±0.11;t=13.870,9.791,7.125;P＜0.05).The absorbance value of lncRNA control group,tumor volume and mass in nude mice[1.99±0.10,(754.50±71.61)cm3,(4.96±0.87)g]was signif-icantly greater than those of lncRNA OIP5-AS1 KD group[1.55±0.12,(419.17±87.59)cm3,(2.96±0.27)g;t=6.934,5.384,7.260;P＜0.05].The number of migratory and invasive cells(145.00±9.75,89.17±5.38)in lncRNA control group was significantly greater than that in lncRNA O1P5-AS1 KD group(110.67±11.52,63.17±10.61;t=5.571,5.353,P＜0.05).The protein expression levels of proliferating cell nuclear antigen(Ki-67),focal adhesion kinase(FAK),mesenchymal cell markers(N-cadherin and Vimentin)inLncRNA control group(1.02±0.11,1.44±0.16,1.21±0.17,1.44±0.16)were significantly higher than in lncRNA OIP5-AS1 KD group(0.50±0.09,0.91±0.09,0.67±0.17,0.69±0.09;t=8.755,7.117,6.509,10.100,P＜0.05).The expression level of epithelial marker(E-cadherin)in lncRNA control group(1.16±0.12)was significantly lower than that in lncRNA OIP5-AS1 KD group(1.78±0.17,t=7.428,P＜0.05).The proportion of G0/G1 phase cells in lncRNA control group[(59.17±4.02)％]was significantly lower than that in lncRNA OIP5-AS1 KD group[(75.17±5.91)％,t=5.481,P＜0.05].The proportion of S phase cells in lncRNA control group[(32.83±5.98)％]was significantly higher than that in lncRNA OIP5-AS1 KD group[(24.50±2.43)％,t=3.162,P＜0.05].Conclusion LncRNA OIP5-AS1 is highly expressed in renal carcinoma cells and is involved in cell biological behaviors such as proliferation and metastasis of renal carcinoma cells.
Long non-coding RNAKidney cancer cellsProliferationMetastasis
NETL
NSTL
万方数据
