长链非编码RNA UCA1通过微小RNA-203对食管癌细胞增殖、凋亡和侵袭的影响

Effects of long noncoding RNA UCA1 on proliferation,apoptosis and invasion of esophageal cancer cells through microRNA-203

目的探讨长链非编码RNA(lncRNA) UCA1通过微小RNA(miRNA, miR)-203对食管癌细胞增殖, 凋亡和侵袭的影响。方法选取河南省人民医院2021年2月到2022年1月手术切除的142例食管癌和癌旁组织, 采用荧光定量聚合酶链式反应(PCR)分析lncRNA UCA1和miR-203表达水平;乳腺癌MCF-7细胞系随机分为lncRNA对照组、短发卡RNA(shRNA) UCA1组、miRNA对照组和miR-203组。采用蛋白质印迹法(Western blot)、流式细胞术和Transwell实验分析不同处理的细胞增殖、凋亡和转移情况。荧光素酶基因报告实验检测lncRNA UCA1和miR-203的关系。蛋白质印迹法(Western blot)分析增殖、凋亡和转移蛋白表达水平。组间计量资料比较采用t检验。结果癌旁组织lncRNA UCA1表达水平(0.89±0.14)低于食管癌组织(2.71±0.20), 差异有统计学意义(t=4.661, P<0.05);癌旁组织miR-203表达水平(1.18±0.16)高于乳腺癌组织miR-203表达水平(0.34±0.11), 差异有统计学意义(t=3.917, P<0.05)。lncRNA对照组细胞48 h吸光度(A)值(2.19±0.21)高于shRNA UCA1组细胞48 hA值(1.47±0.10), 差异有统计学意义(t=3.198, P<0.05)。miRNA对照组细胞48 hA值(2.28±0.24)高于miR-203组细胞48 hA值(1.33±0.17), 差异有统计学意义(t=3.139, P<0.05)。lncRNA对照组细胞凋亡比例[(6.81±0.89)%]低于shRNA UCA1组细胞[(22.09±4.12)%], 差异有统计学意义(t=4.871, P<0.05)。miRNA对照组细胞凋亡比例[(6.09±0.82)%]低于miR-203组细胞[(27.09±3.81)%], 差异有统计学意义(t=5.557, P<0.05)。lncRNA对照组细胞侵袭数量[(132.44±11.82)个]高于shRNA UCA1组细胞侵袭数量[(68.29±6.82)个], 差异有统计学意义(t=5.719, P<0.05)。miRNA对照组细胞侵袭数量[(126.98±10.54)个]高于miR-203组细胞侵袭数量[(74.29±8.71)个], 差异有统计学意义(t=6.209, P<0.05)。lncRNA UCA1有miR-203的结合位点, 起着海绵作用。lncRNA对照组细胞MAP3K1、IRS-1和RGS7 mRNA表达水平(1.39±0.21、1.22±0.17、1.10±0.15)明显高于shRNA UCA1组细胞(0.76±0.19、0.71±0.20、0.49±0.17), 差异有统计学意义(t=3.191、3.018、3.381, P<0.05)。结论 lncRNA UCA1在食管癌中高表达, 通过结合miR-203, 调控着食管癌细胞增殖、凋亡和侵袭过程。

Objective To investigate the effects of long noncoding RNA(lncRNA)UCA1 on the proliferation,apoptosis and invasion of esophageal cancer cells through microRNA(miR)-203.Methods A total of 142 cases of esophageal cancer and adjacent tissues surgically removed in our hospital from February 2021 to January 2022 were selected as research objects.The expression levels of lncRNA UCA1 and miR-203 were analyzed by fluorescence quantitative polymerase chain reaction(PCR).MCF-7 cell lines of breast cancer were randomly divided into lncRNA control group,short hairpin RNA(shRNA)UCA1 group,miRNA control group and miR-203 group.Cell proliferation,apoptosis and metastasis were analyzed by cell counting kit-18(CCK-8)assay,flow cytometry and Transwell experiment.Luciferase gene reporter assay was used to detect the relationship between lncRNA UCA1 and miR-203.The measurement data between groups were compared by t-test.Results The expression level of lncRNA UCA1 in adjacent tissues(0.89±0.14)was significantly lower than that in esophageal cancer tissues(2.71±0.20,t=4.661,P<0.05).The expression level of miR-203 in adjacent tissues(1.18±0.16)was higher than that in esophageal cancer tissues(0.34±0.11,t=3.917,P<0.05).The absorbance(A)value in lncRNA control group(2.19±0.21)was higher than that in shRNA UCA1 group(1.47±0.10,t=3.198,P<0.05).The A value of cells in miRNA control group(2.28±0.24)was lower than that in miR-203 group(1.33±0.17,t=3.139,P<0.05).The apoptosis rate of lncRNA control group[(6.81±0.89)%]was lower than that of shRNA UCA1 group[(22.09±4.12)%,t=4.871,P<0.05].The apoptosis rate in miRNA control group[(6.09±0.82)%]was lower than that in miR-203 group[(27.09±3.81)%,t=5.557,P<0.05].The number of invasive cells in lncRNA control group(132.44±11.82)was greater than that in shRNA UCA1 group(68.29±6.82,t=5.719,P<0.05).The number of invasive cells in miRNA control group[(126.98±10.54)cells]was greater than that in miR-203 group[(74.29±8.71)cells],and the difference was statistically significant(t=6.209,P<0.05).lncRNA UCA1 has the binding site of miR-203 and acts as a sponge.The mRNA expression levels of MAP3K1,IRS-1 and RGS7 in lncRNA control group(1.39±0.21,1.22±0.17,1.10±0.15)were significantly higher than those in shRNA UCA1 group(0.76±0.19,0.71±0.20,0.49±0.17,t=3.191,3.018,3.381,P<0.05).Conclusion LncRNA UCA1 is highly expressed in esophageal cancer and regulates the proliferation,apoptosis and invasion of esophageal cancer cells by binding miR-203.