OPN转染BMSCs对高转移潜能肝癌细胞的影响  

Effect of OPN transfected BMSCs on MHCC97-H

作　　者：李天然[1] 蔡立杰[1] 赵绍宏[2] 魏正茂[3] 霍天龙[3] 卢光明 宋斌[1] 

机构地区：[1]福建医科大学福总临床医学院第一附属医院放射科,福建莆田351100 [2]解放军总医院第一附属医院放射科 [3]北京大学人民医院放射科 [4]南京总医院放射科

出　　处：《胃肠病学和肝病学杂志》2015年第9期1057-1061,共5页Chinese Journal of Gastroenterology and Hepatology

基　　金：国家自然科学基金资助项目(81271607);南京军区医药卫生重点项目资助(111Z035);福建省自然基金资助项目(2013J0I392);国家博士后基金(2015M572810)

摘　　要：目的观察经骨桥蛋白(osteopontin,OPN)转染人骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)对高转移潜能肝癌细胞(high metastatic potential hepatocellular carcinoma cells,MHCC97-H)增殖、侵袭能力的影响。方法构建OPN慢病毒载体,转染BMSCs。CCK-8法检测与BMSCs共培养前后MHCC97-H的增殖能力,共培养48 h,检测OD值。Transwell法检测BMSCs与MHCC97-H细胞共培养后细胞侵袭情况,共培养48 h,结晶紫染色,细胞计数。PCR法检测共培养前后MHCC97-H转移相关因子OPN、αV、β3、基质金属蛋白酶-2(MMP-2)基因表达。结果 BMSCs基因转染成功,OPN高表达。MHCC97-H与BMSCs细胞共培养后,MHCC97-H增殖能力增强,与对照组比较,差异有统计学意义(P<0.05)。经OPN转染的BMSCs细胞对MHCC97-H细胞增殖无明显作用(P>0.05)。经OPN转染的BMSCs细胞对MHCC97-H细胞侵袭具有明显的促进作用(P<0.05)。经OPN转染的MHCC97-H细胞OPN表达明显降低,而αv、β3、MMP-2表达明显升高(P<0.01)。结论 BMSCs具有促进MHCC97-H细胞增殖的作用,OPN基因转染的BMSCs细胞具有促进MHCC97-H细胞侵袭的作用。外源性OPN抑制内源性OPN表达,外源性OPN通过激活MMP-2活性促进肿瘤侵袭。Objective To observe the proliferation and invasion ability of osteopontin （OPN） transfected bone mar- row mesenehymal stem cells （BMSCs） on high metastatic hepatoeelluar carcinoma cells （MHCC97-H）. Methods OPN lentiviral vector was builded and BMSCs were transfeeted. Proliferative capacity of MHCC97-H co-culture with BMSCs test was detected by CCK-8. After 48 hours, OD value was detected. The invasion ability of MHCC97-H cells after BM- SCs with hepatoma ceils co-culture experiments was detected by Transwell. After 48 hours co-culture, crystal violet was stained and cells were counted. Metastatic factors, such as OPN、αV、β3、 matrix metalloproteinase-2 （MMP-2） gene expressions of MHCC97-H cells were detected by PCR mothods. Results BMSCs transfecting successed, OPN showed high expression. Proliferation ability of MHCC97-H co-cultured with BMSCs cells was increased and there was statisti- cally different compared with the control group （P 〈 0.05）. OPN gene transfeeted BMSCs had no significant effect on proliferation ability of MHCC97-H. Invasion ability of MHCC97-H co-cultured with gene infected BMSCs cells was in- creased （P 〈 0. 05 ）. OPN gene expression of OPN gene transfeeted BMSCs was significantly lower and OPN、αV、β3、 MMP-2 gene expressions were significantly higher compared with control group （P 〈 0. 01 ）. Conclusion BMSCs can promote MHCC97-H cells proliferation, and OPN gene transfected BMSCs can promote MHCC97-H cells invasion. Exogenous OPN can inhibit endogenous OPN expression. Exogenous OPN can activate MMP-2 activity and promote tumor invasion.

关 键 词：骨髓间充质干细胞 整合素 骨桥蛋白 高转移潜能肝癌细胞 

分 类 号：R735.7[医药卫生—肿瘤]