转化医学杂志2017，Vol.6Issue(1)：6-11,15,7.DOI:10.3969/j.issn.2095-3097.2017.01.002
肌细胞增强因子2D通过负向调控有丝分裂诱导基因6的表达促进肝癌细胞PLC/PRF5和SMMC7721的增殖和迁移
Myocyte enhancer factor 2D promotes the proliferation and migration of hepatoma cells PLC/PRF5 and SMMC7721 by negatively regulating mitogen-induced gene 6 expression
摘要
Abstract
Objective To study the effects of myocyte enhancer factor 2D ( MEF2D) on prolifera-tion and migration of hepatoma cells PLC/PRF5 and SMMC7721, and regulating the expression of mitogen-induced gene 6 ( MIG6) . Methods The mRNA levels of MEF2D and MIG6 were detected by real time fluorescence quantitative PCR in hcclm3, SMMC7721, PLC/PRF5, HepG2, 7703, Huh7, Bel-7402 cell lines and we drew a linear correlation diagram with “MEF2D mRNA expre-ssion” on the horizontal axis and “MIG6 mRNA expression” on the vertical one in these hepatoma cell lines. The expression of MEF2D and MIG6 were down-regulated by small interfering RNA (siRNA), then the proliferation and migration were observed by 7Sea-Cell Counting Kit ( 7Sea-CCK ) and wound healing assay in PLC/PRF5 cell lines. The expression of MEF2D and MIG6 were detected by western blot after MEF2D down-regulated by siRNA in PLC/PRF5 cell lines or up-regulated by plasmid in SMMC7721 cell lines. The experimental were divided into seven groups:negative control ( NC) group, siMIG6 group, siMEF2D group, si2M ( both MEF2D and MIG6 were down-regulated) group , blank group, empty vector plasmid group and MEF2D group. Results We found that the level of MEF2D mRNA was negatively correlated with expression of MIG6 mRNA in hcclm3, SMMC7721, PLC/PRF5, HepG2, 7703, Huh7, Bel-7402 cell lines ( r=-0. 78) . Next, the results analyzed by 7Sea-CCK showed that the proliferation ability was significantly enhanced at 72 h after siMIG6 treatment compared with NC group in PLC/PRF5 cell lines (OD=3. 73±0. 18, t=3. 84, P=0. 02) , while there was no change at 24 h and 48 h. However, the proliferation ability was signifi-cantly suppressed at 72 h under MEF2D knockdown condition in PLC/PRF5 cell lines compared NC group ( OD=1. 79 ± 0. 22, t=3. 95, P=0. 02 ) , but the proliferation ability was significantly in-creased at 72 h under MEF2D and MIG6 both knockdown condition compared with siMEF2D group (OD=2. 99±0. 03, t=4. 83, P<0. 01). Furthermore, wound healing assay revealed that the cell mobility in siMIG6 group was significantly increased [cell mobility=(51±4)%, t=3. 22, P<0. 05] at 48 h compared with NC group and the cell mobility in siMEF2D group was significantly reduced [cell mobility=(19±3), t=7. 70, P<0. 01] in PLC/PRF5 cell lines, but the cell mobility in si2M group was significantly increased at 48 h compared with siMEF2D group [cell mobility=(46±3)%, t=6. 99, P<0. 01] . The result of western blot showed that, compared with NC group, the expression of MIG6 was significantly increased under MEF2D siRNA treatment in PLC/PRF5 cell lines ( t=7. 86, P<0. 001), while its level was significantly decreased by the elevated level of MEF2D in SMMC7721 cell lines( t=4. 61, P=0. 004) . Conclusion The expression of MEF2D was negatively correlated with the level of MIG6 of PLC/PRF5 and SMMC7721 cell lines. Thus, MEF2D promotes the proliferation and migration of PLC/PRF5 and SMMC7721 cell lines possibly through negatively regulating MIG6 expression.关键词
肌细胞增强因子2D/有丝分裂诱导基因6/肝细胞癌/负向调控/细胞增殖/细胞迁移Key words
Myocyte enhancer factor 2D ( MEF2D)/Mitogenin-duced gene 6 ( MIG6)/Hepatocellular carcinoma/Negative regulation/Cell proliferation/Cell migration分类
生物科学引用本文复制引用
陈潇,林志娟,李孟鹏,陈超,王建勋,刘佳,李培峰..肌细胞增强因子2D通过负向调控有丝分裂诱导基因6的表达促进肝癌细胞PLC/PRF5和SMMC7721的增殖和迁移[J].转化医学杂志,2017,6(1):6-11,15,7.基金项目
国家自然科学基金青年项目(81502065)；山东省自然科学基金青年项目(ZR2014HQ009)；中国博士后科学基金特别资助项目(2016T90613)；中国博士后科学基金面上项目(2015M580574)；山东省博士后创新项目(201602037)；青岛市自主创新计划应用基础研究项目(16-5-1-56-JCH)；青岛市博士后应用研究项目(2015167) （81502065）
