免疫检查点分子B7-H3对骨肉瘤细胞增殖和转移的影响  

Impact of immune checkpoint B7-H3 on proliferation and metastasis of osteosarcoma cells

作　　者：吴晗[1] 吴学建[1] Wu Han;Wu Xuejian(Department of Orthopedics,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]郑州大学第一附属医院骨科,450052

出　　处：《中华实验外科杂志》2020年第11期2039-2041,共3页Chinese Journal of Experimental Surgery

摘　　要：目的观察免疫检查点分子B7-H3过表达对骨肉瘤细胞系MG63、U2OS、K7M2增殖和转移的影响。方法通过慢病毒介导的基因过表达技术,构建骨肉瘤细胞系MG63、U2OS、K7M2的对照组(空载体组)和实验组(B7-H3过表达组)细胞。在体外环境下,应用氮兰四唑盐/吩嗪硫酸甲酯(MTS/PMS)实验和克隆形成实验检测细胞增殖能力;应用划痕实验,检测细胞迁移能力。使用免疫健全的BALB/c小鼠,构建皮下成瘤模型和肺转移模型,检测体内环境下骨肉瘤细胞增殖和转移能力。应用Graph Pad 8统计软件分析,组间比较采用t检验分析。结果MTS/PMS实验结果显示,培养6 d后,MG63、U2OS、K7M2实验组细胞吸光度(A)值(2.466±0.229、2.244±0.163、2.404±0.241)均比对照组(1.821±0.189、1.929±0.164、2.008±0.111)显著增高,差异有统计学意义(t=4.852、3.040、3.338,P<0.05);克隆形成实验结果显示,在培养10 d后,MG63、K7M2实验组细胞形成的克隆数[(24.333±4.163)、(32.667±6.028)个]明显高于对照组[(6.667±2.517)、(15.333±3.215)个],差异有统计学意义(t=6.290、4.395,P<0.05),而U2OS实验组与对照组细胞形成的克隆数[(20.333±3.786)个比(19.667±3.055)个],差异无统计学意义(t=0.237,P>0.05)。划痕实验结果显示,3种骨肉瘤细胞实验组与对照组比较,在划痕愈合率方面的差异均无统计学意义。小鼠皮下成瘤模型表明,过表达B7-H3在体内环境下可以显著促进骨肉瘤的增殖。小鼠肺转移模型表明,过表达B7-H3在体内环境下可以显著促进骨肉瘤的转移。结论过表达B7-H3在体外环境可部分增强骨肉瘤细胞的增殖,而在体内环境下可显著促进骨肉瘤的形成和转移。Objective To observe the function of overexpressed B7-H3 on proliferation and metastasis of MG63,U2OS and K7M2 osteosarcoma cell lines.Methods By infecting with recombinant lentivirus,the stable cell lines of control group(vector group)and B7-H3(overexpressing group)were constructed respectively.Cell proliferation was detected with 3-(4,5-Dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium(MTS)/Phenazine methosulfate(PMS)assay and colony formation in vitro,wound healing assay was conducted to verify the effect of B7-H3 on cell migration.With immune competent BALB/c mouse,the subcutaneous tumor model and pulmonary metastasis model were established,and the impact of B7-H3 on tumor formation and metastasis of osteosarcoma was further assessed in vivo.Results The results of MTS/PMS assay showed that after 6 days,absorbance(A)values of B7-H3 groups in MG63,U2OS,and K7M2 cell lines(2.466±0.229,2.244±0.163,2.404±0.241)was significantly higher than that of control group(1.821±0.189,1.929±0.164,2.008±0.111,t=4.852,3.040,3.338,P<0.05).Colony formation showed that after culturing for 10 days,the number of clones formed by B7-H3 groups in MG63 and K7M2 cell lines(24.333±4.163,32.667±6.028)was significantly higher than that of control groups(6.667±2.517,15.333±3.215,t=6.290,4.395,P<0.05),while difference between groups of U2OS cell lines were not statistically significant(20.333±3.786 vs.19.667±3.055,t=0.237,P>0.05).Wound healing assay showed that,there was no statistically significant difference in healing rates,between three B7-H3 groups and three control groups,respectively.The subcutaneous tumor model showed that overexpressed B7-H3 could significantly promote the proliferation and tumor formation of osteosarcoma in vivo.The metastasis model showed that overexpressed B7-H3 could significantly enhance the metastasis of osteosarcoma in vivo.Conclusion Overexpressed B7-H3 partly promoted the proliferation of osteosarcoma in vitro,but significantly contributed to osteosarcoma formatio

关 键 词：骨肉瘤 免疫检查点 B7-H3 增殖 转移 

分 类 号：R738.1[医药卫生—肿瘤]