长链非编码RNA Opa相互作用蛋白5-反义转录物1在结直肠腺癌组织的表达及其与细胞增殖和转移的关系

Expression of long non-coding RNA OIP5-AS1 in colorectal cancer and its relationship with cell proliferation and metastasis

目的探讨长链非编码RNA Opa相互作用蛋白5-反义转录物1(lncRNA OIP5-AS1)在结直肠癌组织中的表达及其与结直肠癌细胞增殖和转移的关系。方法选取2023年2月至2024年2月期间于河南省驻马店市中心医院和新乡医学院第一附属医院就诊并行结直肠癌根治术的51例结直肠癌患者,荧光定量聚合酶链反应(PCR)法检测所有患者结直肠癌组织和癌旁组织中lncRNA OIP5-AS1的mRNA表达水平;将短发卡RNA(shRNA)空载体(shRNA-Control)、OIP5-AS1敲低质粒(shRNA-OIP5-AS1)转染至HCT116结直肠癌细胞中,采用噻唑蓝(MTT)比色法检测shRNA-Control和shRNA-OIP5-AS1细胞的增殖活力;采用Transwell实验检测shRNA-Control和shRNA-OIP5-AS1细胞的迁移和侵袭能力;采用酶联免疫吸附试验(ELISA)检测两组细胞中波形蛋白(Vimentin)、N-钙黏蛋白(N-Cadherin)、锌指E-盒结合同源盒蛋白1(ZEB 1)、E-钙黏蛋白(N-Cadherin)的蛋白表达,组间计量数据采用独立样本t检验。结果lncRNA OIP5-AS1 mRNA在结直肠癌组织(1.50±0.13)中的表达水平明显高于癌旁组织(0.54±0.08),差异有统计学意义(t=45.99,P<0.05)。shRNA-OIP5-AS1细胞(0.80±0.06)的吸光度值明显低于shRNA-Control细胞(1.30±0.04),差异有统计学意义(t=16.62,P<0.05)。shRNA-OIP5-AS1细胞迁移细胞数[(74.50±3.73)个]和侵袭细胞数[(66.50±5.09)个]明显低于shRNA-Control细胞[(126.33±6.28)、(109.67±3.88)个],差异有统计学意义(t=17.38、16.52,P<0.05)。shRNA-OIP5-AS1细胞E-Cadherin[(5.36±0.28)ng/ml]蛋白表达水平明显高于shRNA-Control细胞[(2.96±0.13)ng/ml],差异有统计学意义(t=18.97,P<0.05)。shRNA-OIP5-AS1细胞N-Cadherin[(231.02±27.11)pg/ml]、Vimentin[(5.61±0.40)ng/ml]和ZEB1[(2.59±0.28)ng/ml]蛋白表达水平明显低于shRNA-Control细胞[(656.48±39.49)pg/ml、(8.31±0.36)ng/ml、(6.04±0.26)ng/ml],差异有统计学意义(t=21.76、12.32、22.12,P<0.05)。结论lncRNA OIP5-AS1在结直肠癌组织中表达水平上调,敲低结直肠癌细胞lncRNA OIP5-AS1的表达可通过抑制细胞的上皮-间充质转化,从而减缓结直肠癌细胞的增殖、转移和侵袭。

ObjectiveTo investigate the expression of long non-coding RNA(lncRNA)OIP5-AS1 in colorectal cancer and its relationship with cell proliferation and metastasis.MethodsTotally,51 patients with colorectal cancer who received radical resection in Zhumadian Central Hospital and the First Affiliated Hospital of Xinxiang Medical College,Henan Province from February 2023 to February 2024 were selected as the study objects.The mRNA expression of lncRNA OIP5-AS1 in colorectal cancer tissues and adjacent tissues was detected by fluorescence quantitative polymerase chain reaction(PCR).The short hairpin RNA empty vector(shRNA-control)and OIP5-AS1 knockdown plasmid(shRNA-OIP5-AS1)were transfected into HCT116 colorectal cancer cells,and the proliferative activity of shRNA-control and shRNA-OIP5-AS1 was detected by methyl thiazolyl tetrazolium(MTT)colorimetric method.Transwell assay was used to detect the migration and invasion ability of shRNA-control and shRNA-OIP5-AS1 cells.The protein expression levels of Vimentin,N-Cadherin,ZEB1 and E-cadherin were detected by enzyme linked immunosorbent assay(ELISA).Independent sample t test was used for inter-group measurement data.ResultsThe mRNA expression of lncRNA OIP5-AS1 in colorectal cancer tissues(1.50±0.13)was significantly higher than that in para-carcinoma tissues(0.54±0.08,t=45.99,P<0.05).The absorbance of shRNA-OIP5-AS1 cells(0.80±0.06)was significantly lower than that of shRNA-control cells(1.30±0.04,t=16.62,P<0.05).The number of migrating cells[(74.50±3.73)]and invasion cells[(66.50±5.09)]of shRNA-OIP5-AS1 cells was significantly less than that of shRNA-control cells[(126.33±6.28),(109.67±3.88),t=17.38,16.52,P<0.05].The expression of E-Cadherin[(5.36±0.28)ng/ml]in shRNA-OIP5-AS1 cells was significantly higher than that in shRNA-control cells[(2.96±0.13)ng/ml,t=18.97,P<0.05).The protein expression of N-Cadherin[(231.02±27.11)pg/ml],Vimentin[(5.61±0.40)ng/ml]and ZEB1[(2.59±0.28)ng/ml]in shRNA-OIP5-AS1 cells was significantly lower than that in shRNA-control cells[(656.48±39.49)pg/ml,(8.31±0.36)ng/ml,(6.04±0.26)ng/ml,t=21.76,12.32,22.12,P<0.05].ConclusionThe lncRNA OIP5-AS1 expression is up-regulated in colorectal cancer.Knocking down the expression of lncRNA OIP5-AS1 can slow down the proliferation,metastasis and invasion of colorectal cancer cells by inhibiting the epithelial mesenchymal transformation of cells.