摘要
目的通过观察在miR-21作用下hTERT的表达情况,探讨在宫颈癌HeLa细胞中miR-21对hTERT的调控作用。方法利用RT-PCR法及West Blot法分别检测HeLa中miR-21的表达,以及hTERT蛋白的表达情况;通过实验设计,生成miR-21模拟物和miR-21抑制剂,并分别设立两组的空白对照序列;将空转liposome2000(lipo2000)设为对照组,利用脂质体liposome2000包裹宫颈癌细胞株HeLa并予以转染。在转染后24 h及时检测hTERT及蛋白的表达量。结果HeLa细胞株中miR-21及hTERT表达阳性;转染后24 h模拟物组、模拟物对照组、抑制剂组、抑制剂对照组、脂质体组hTERT mRNA相对表达水平分别为(0.51±0.33)、(0.69±0.19)、(1.33±0.39)、(0.83±0.26)和(0.58±0.16);蛋白相对表达水平分别为(0.41±0.18)、(0.62±0.18)、(1.45±0.60)、(0.83±0.15)和(0.82±0.30);模拟物组与其对照组及脂质体组相比较,hTERT mRNA及蛋白表达情况均较低且差异有统计学意义(P<0.05);相反,在抑制剂组与其对照组及脂质体组相比较中,hTERT mRNA及蛋白表达情况明显升高,差异有统计学意义(P<0.05)。结论通过检测发现miR-21及hTERT基因在HeLa细胞株中均呈现高表达状态;miR-21可以调节hTERT mRNA和蛋白表达水平,从而发挥负向调控hTERT的作用。
Objective To observe the effect of miR-21 on hTERT expression in human cervical cancer Hela cell line,explore the regulatory effect of miR-21 on hTERT.Methods RT-PCR was used to detect the expressions of miR-21 and hTERT mRNA in human cervical cancer Hela cell line.West Blot was used to detect the expression of hTERT protein;miR-21 mimics,miR-21 inhibitor,mimics control and inhibitor control were designed and synthesized,then they were transfected into human cervical cancer Hela cell line after parceled by liposome 2000,blank liposome 2000 was designed as control group.The changes of expressions of hTERT mRNA and protein at 24 hours after transfection were detected.Results miR-21 and hTERT were positive in human cervical cancer Hela cell line;the relative expression levels of hTERT mRNA in mimics group,mimics control group,inhibitor group,inhibitor control group and liposome group at 24 hours after transfection were(0.51±0.33),(0.69±0.19),(1.33±0.39),(0.83±0.26)and(0.58±0.16),respectively;the relative expression levels of h TERT protein were(0.41±0.18),(0.62±0.18),(1.45±0.60),(0.83±0.15)and(0.82±0.30),respectively.The relative expression levels of hTERT mRNA and protein in mimics group were statistically significantly lower than those in mimics control group and liposome group(P<0.05).The relative expression levels of hTERT mRNA and protein in inhibitor group were statistically significantly higher than those in inhibitor control group and liposome group(P<0.05).Conclusions miR-21 and hTERT genes are highly expressed in HeLa cell lines.MiR-21 can regulate hTERT mRNA and protein expression levels,thus negatively regulating hTERT.
作者
刘洋
廖婧
卢又汇
孙春意
孟昱时
LIU Yang;LIAOJing;LU You-hui;SUN Chun-yi;MENG Yu-shi(Dept.of Reproductive Medicine,The Second Affiliated Hospital of Kunming Medical University,Yunnan Kunming 650101,China;Dept.of Gynecology,The Second Affiliated Hospital of Kunming Medical University,Yunnan Kunming 650101,China)
出处
《昆明医科大学学报》
CAS
2021年第3期23-28,共6页
Journal of Kunming Medical University
基金
国家自然科学基金资助项目(81860515)
云南省科技厅-昆明医科大学应用基础研究联合专项基金资助项目[2018FE001(-055)]
云南省万人计划“青年拔尖人才”培养基金资助项目(云人社发[2018]73号)
云南省万人计划“名医专项”培养基金资助项目(云厅字[2018]11号)
云南省高层次卫生健康技术人才“妇产科学学科带头人”(D-2019004)
