长链非编码RNA H19靶向Bcl-2的表达调控宫颈癌细胞侵袭转移行为的机制  

Mechanism of Long-chain non-coding RNA H19 Targeting Bcl-2 Expression for Regulating Invasion and Metastasis of Cervical Cancer Cells

作　　者：于洁[1] 卢博奇[1] 朱丽业[1] 刘宁[1] YU Jie;LU Bo-qi;ZHU Li-ye;LIU Ning(Department of Gynecology and Obstetrics,Maternal and Child Health Hospital of Haidian Region,Beijing 100080,China)

机构地区：[1]北京市海淀区妇幼保健院妇产科,北京100080

出　　处：《解放军医药杂志》2019年第1期12-17,共6页Medical & Pharmaceutical Journal of Chinese People’s Liberation Army

基　　金：北京市海淀区科学技术委员会专项项目(K200684)

摘　　要：目的探讨长链非编码RNA(IncRNA) H19调控宫颈癌细胞的侵袭转移行为及其机制。方法选择2016年7月—2017年1月手术切除并且经病理检查确诊为宫颈癌的组织50例,同时取癌旁正常组织50例。选择不同类型人宫颈癌细胞株。q PCR检测H19在宫颈癌组织和不同宫颈癌细胞株中的表达情况;分析H19表达与宫颈癌患者临床病理参数间的关系;免疫共沉淀实验检测H19与Bcl-2的相互作用。将宫颈癌细胞分为3组,空白对照组使用未经处理的宫颈癌细胞,阴性对照组使用空白慢病毒处理的宫颈癌细胞,H19-siRNA组使用H19-siRNA慢病毒处理的宫颈癌细胞。划痕愈合实验检测抑制H19后3组宫颈癌细胞迁移能力变化情况,Transwell侵袭实验检测抑制H19后3组宫颈癌细胞的侵袭行为变化情况;蛋白免疫印迹法检测3组lncRNA H19对E-钙黏蛋白(E-Cadherin)和Bcl-2蛋白的调控情况。结果与正常宫颈组织相比,宫颈癌组织中H19的表达水平升高,且其表达水平与宫颈癌的病理分期及淋巴结转移情况有关(P <0. 05)。与其他宫颈癌细胞株相比,He La细胞中H19表达水平最高(P <0. 05),作为实验的细胞株。H19-siRNA组在36 h和72 h宫颈癌细胞迁移率低于空白对照组和阴性对照组,通过Matrigel基质胶的细胞数量少于空白对照组和阴性对照组(P <0. 05)。H19-siRNA组Bcl-2表达水平低于空白对照组和阴性对照组(P <0. 05),而3组E-Cadherin表达水平比较差异无统计学意义(P> 0. 05)。结论 H19能调控宫颈癌细胞Bcl-2的表达,抑制H19的表达后可以抑制宫颈癌细胞的增殖和侵袭能力。H19可以调控Bcl-2的表达影响宫颈癌细胞的生物学行为。Objective To investigate effect of long-chain non-coding RNA(IncRNA)H19 regulating invasion and metastasis of cervical cancer cells and its mechanism.Methods A total of 50 cases of cervical cancer tissues from patients who were confirmed by pathological examination after surgical ablation during July 2016 and January 2017 were selected,and other 50 cases of adjacent normal tissues were also selected simultaneouly.Different types of human cervical cancer cell lines were selected.Quantitative polumerase chain reaction(qPCR)was used to detect H19 expression in cervical cancer tissues and different types of cervical cancer cell lines.Correlations between H19 expression and clinicopathological parameters in patients with cervical cancer were analyzed.Immunoprecipitation assay was used to detect interaction between H19 and Bcl-2.Cervical cancer cells were divided into three groups.Untreated cervical cancer cells were used in blank control group,and cervical cancer cells treated by blank lentivirus were used in negative control group,and cervical cancer cells treated by H19-siRNA lentivirus were used in H19-siRNA group.Among three groups,scratch healing test was used to detect changes of migration ability of cervical cancer cells after inhibiting H19;Transwell invasion assay was used to detect changes of invasive behavior of cervical cancer cells after inhibiting H19;western blotting was used to detect regulation effects of lncRNA H19 on E-cadherin and Bcl-2 proteins.Results Compared with those in normal cervical tissues,in cervical cancer tissues,H19 expression was significantly higher,and the expression was related to pathological staging of cervical cancer and lymph nodes metastasis(P<0.05).Compared with those in other cervical cancer cell lines,H19 expression was the highest in HeLa cells(P<0.05),and they were used as experimental cell lines,In H19-siRNA group,migration rates of cervical cancer cells at 36 h and 72 h after cultivation were significantly lower,while cell number of passing matrigel was less than those

关 键 词：宫颈肿瘤 H19 BCL-2 细胞增殖 肿瘤浸润 

分 类 号：R737.33[医药卫生—肿瘤]