HHLA2过表达胃癌细胞株构建及细胞功能的初步研究  

Construction and cell function study of HHLA2-overexpressing gastric cancer cells

作　　者：韩晓娟 徐佳倩 朱玉兰 王莹 李源 冯珺 邵东[1] Xiaojuan Han;Jiaqian Xu;Yulan Zhu;Ying Wang;Yuan Li;Jun Feng;Dong Shao(Department of Gastroenterology,the Third Affiliated Hospital of Soochow University,Changzhou 213003,China;Department of Oncology,the Third Affiliated Hospital of Soochow University,Changzhou 213003,China;Cancer Biological Diagnosis and Treatment Center,the Third Affiliated Hospital of Soochow University,Changzhou 213003,China)

机构地区：[1]苏州大学附属第三医院消化内科,江苏常州213003 [2]苏州大学附属第三医院肿瘤科,江苏常州213003 [3]苏州大学附属第三医院肿瘤生物诊疗中心,江苏常州213003

出　　处：《中华消化病与影像杂志（电子版）》2023年第6期373-377,共5页Chinese Journal of Digestion and Medical Imageology(Electronic Edition)

基　　金：常州市应用基础研究计划(CJ20210074);常州市卫生健康委青年人才科技项目(QN202015)。

摘　　要：目的使用慢病毒载体构建稳定过表达人内源性逆转录病毒H-长末端重复关联蛋白2(HHLA2)的胃癌细胞系,探讨细胞发生的生物学功能改变。方法由目的基因以及Lipo2000转染试剂构建重组质粒,将该质粒转染293T细胞,浓缩其细胞上清液并测定浓度,再将浓缩液转染胃癌细胞株BGC-823和MGC80-3,利用流式细胞术分选并验证GFP+稳定过表达的细胞株。开展细胞功能实验(含迁移试验和增殖试验等)以分析过表达HHLA2胃癌细胞株的生物学功能。结果酶切及测序结果示,重组慢病毒载体成功构建。流式分选及验证示,HHLA2过表达胃癌稳转株。过表达HHLA2可使胃癌细胞株MGC80-3和BGC-823加快增殖,其24 h、48 h、72 h吸光度值均较对照细胞株升高(P<0.05)。过表达HHLA2可增强胃癌细胞株BGC-823的迁移能力,其24 h、48 h迁移率均较对照细胞株升高(P<0.05);MGC80-3细胞迁移距离未明显变长(P>0.05)。结论HHLA2过表达能够提高胃癌细胞的增殖和迁移能力。Objective To construct gastric cancer cell lines overexpressing human endogenous retrovirus subfamily H long terminal repeat associating protein 2(HHLA2)by lentivirus vector and verify their biological functions.Methods The target gene and Lipo2000 reagent were used to construct recombinant plasmid,which was then applied to transfect 293T cell.We collect The cell supernatant was concentrated and the concentration was determined.The concentrated liquid was then transfected into gastric cancer cell lines BGC-823 and MGC80-3.The flow cytometry was used to sort and verify the GFP+stable overexpressing cell lines.Cell function experiments(including migration experiment and proliferation experiment)were applied to explore the biological functions of overexpressing HHLA2 gastric cancer cell lines.Results The results of restriction enzyme digestion and sequencing showed that the recombinant lentivirus vector was successfully constructed.Flow sorting and verification showed that HHLA2 overexpressed gastric cancer stably transformed strains.Overexpression of HHLA2 could accelerate the proliferation of gastric cancer cell lines MGC80-3 and BGC-823,and their absorbance values at 24 h,48 h and 72 h were higher than those of the control cell lines(P<0.05).Overexpression of HHLA2 could enhance the migration ability of gastric cancer cell line BGC-823,and its migration rate at 24 h and 48 h was higher than that of the control cell line(P<0.05).However,the migration distance of MGC80-3 cells did not increase significantly(P>0.05).Conclusion Overexpression of HHLA2 can promote the proliferation and migration ability of gastric cancer.

关 键 词：胃癌 人内源性逆转录病毒-H长末端重复关联蛋白2 慢病毒载体 流式细胞术 

分 类 号：R735.2[医药卫生—肿瘤]