摘要
目的探讨脑源性神经营养因子反义长链非编码RNA(brain-derived neurotrophic factor antisense long noncoding RNA,BDNF AS)对人肝癌细胞增殖和迁移能力的影响。方法采用BDNF AS过表达质粒及空载体瞬时转染肝癌细胞株HepG2和MHCC 97-H,采用实时PCR方法检测BDNF AS在肝癌细胞中的表达水平,采用3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium(MTS)方法检测细胞增殖情况,采用碘化丙啶(propidium iodide,PI)染色流式细胞术检测细胞周期变化,采用划痕实验和transwell小室评价细胞迁移能力的变化。结果 HepG2和MHCC 97-H细胞转染BDNF AS过表达质粒后BDNF AS和mRNA表达均上调(均P<0.01)。转染第4天,细胞增殖均受到抑制(均P<0.05)。转染48 h后,HepG2和MHCC 97-H细胞均出现S期阻滞(均P<0.01),细胞的迁移能力均下降(均P<0.01)。结论 BDNF AS过表达抑制肝癌细胞的增殖和迁移,可能对肝癌细胞的生长运动具有重要的负性调控作用。
Objective To investigate the effects of antisense long noncoding RNA of brain-derived neurotrophic factor（BDNF AS） on the proliferation and migration of human hepatocellular carcinoma cell lines. Methods Human hepatocellular carcinoma Hep G2 and MHCC 97-H cells were transiently transfected with BDNF AS over-expression plasmid and the empty vector. Real-time PCR was used to examine the expression level of BDNF AS in Hep G2 and MHCC 97-H cells.Cell proliferation was examined by 3-（4,5-dimethylthiazol-2-yl）-5-（3-carboxymethoxyphenyl）-2-（4-sulfophenyl）-2 H-tetrazolium（MTS）,and cell cycle was studied by propidium iodide（PI） staining using flow cytometry. The wound healing and transwell assay were used to evaluate cell migration. Results After transfection with BDNF AS over-expression plasmid,the expression of BDNF AS and mRNA were up-regulated in Hep G2 and MHCC 97-H cells（all P〈0. 01）. Cell proliferation was inhibited 4 d after transfection（both P〈0. 01） and the proportion of S phase cells was increased 48 h after transfection（both P〈0. 01）. Moreover,the migratory ability of transfected cells was also inhibited 48 h after transfection（both P〈0. 01）. Conclusion BDNF AS over-expression inhibits the proliferation and migration of liver cancer cells,indicating that BDNF AS may have a negative regulatory role in the development and mobility of hepatocellular carcinoma.
作者
郭大伟
周品一
赵旭鹏
初晨
江宏伟
周勇
Guo Dawei;Zhou Pinyi;Zhao Xupeng;Chu Chen;Jiang Hongwei;Zhou Yong(The Fourth Department of General Surgery, The Fourth Affiliated Hospital of China Medical University, Liaoning 110032,China)
出处
《实用肿瘤杂志》
CAS
2018年第2期116-121,共6页
Journal of Practical Oncology
基金
辽宁省自然科学基金(2015020739)
