乙型肝炎表面S+前S1融合基因转化CHO-dhfr^-的细胞染色体变化及致瘤性  

Chromosome aberration and carcinogenicity of CHO dhfr ~ - cells transformed by plasmid containing S+ and Pre S1 of fussion gene of hepatitis B virus

作　　者：田淑芳[1] 阮薇琴[1] 王秀平[1] 张陵林[1] 杨芙蓉[1] 宗芳[1] 阮力[1] 

机构地区：[1]中国预防医学科学院病毒学研究所遗传免疫室,北京100052

出　　处：《中华实验和临床病毒学杂志》2001年第3期208-211,共4页Chinese Journal of Experimental and Clinical Virology

基　　金：8 6 3高科技发展计划资助项目 (86 3 2 18 0 1)

摘　　要：目的　了解乙型肝炎病毒表面S +前S1融合重组DNA(乙型肝炎重组DNA)整合到中国地鼠卵巢细胞二氢叶酸还原酶阴性突变株细胞 (CHOdhfr- )染色体的变化及致瘤性。方法　应用分子克隆技术构建含HBsAg的表面抗原S +前S1基因的质粒经转染、克隆、加压 (MTX和MSX)筛选出高效表达乙型肝炎表面S +S1融合抗原的转基因细胞株 (GdSS1 18)进行染色体制片 ,同时将制备的细胞、细胞DNA和细胞匀浆皮下接种裸鼠观察 3周。结果　整合了外源乙型肝炎重组DNA的转基因细胞系 (GdSS1 18)不同的传代均发生染色体结构畸变 ,畸变率为 11% ,5 6%及 2 9% ,而未整合外源乙型肝炎重组DNA的对照CHOdhfr- 细胞为 6% ,二者的染色体众数无明显变化 ,均为 2 0条。整合与未整合外源重组DNA的细胞接种裸鼠以后观察 3周均无肿瘤生长。结论　整合有外源乙型肝炎重组DNA的细胞株 (GdSS1 18)其染色体结构畸变率高于未整合的CHO dhfr- 对照细胞 ,但二者的染色体众数仍为 2 0条 ;不同代数GdSS1 18细胞产物接种裸鼠后未见肿瘤的生长。Objective To investigate the chromosome aberrations and carcinogenicity of CHO dhfr - cell induced by integration of plasmid containing S+ and Pre S1 fusion gene of hepatitis B surface antigen(HBsAg).Methods The plasmid pCHBSS1G was constructed with S+ Pre S1 of HBsAg.CHO dhfr - cells were transformed with this recombinant plasmid DNA and CHO cells lines with integrated DNA were cloned and named GdSS1 18.The GdSS1 18 cell lines secreting the S+ Pre S1 fusion protein of HBsAg at high level were developed by screening in increased concentration of MTX and MSX in cultured media.To make sample of cells chromosome,the cells integrated DNA were subcutaneously injected into nude mouse.Results The CHO dhfr - cell lines integrated with S and Pre S1 fusion protein of HBsAg were developed and named GdSS1 18 cell lines.The frequencies and type of chromosome aberrations of the GdSS1 18 cell lines with different passage generations were 11%,56% and 29%,respectively,while that of the control CHO dhfr - cell lines was 6%.There was no change in the mode of chromosomes,both cell lines having 20 chromosomes.Both cell lines were non oncogenic in nude mouse.Conclusion The chromosome aberration of CHO dhfr - cells with integrated DNA were obviously higher than that of the original CHO dhfr - cells without integrated DNA.Both cell lines were non oncogenic in nude mouse.

关 键 词：乙型肝炎表面抗原 仓鼠 卵巢 染色体畸变 致瘤性 

分 类 号：R392-33[医药卫生—免疫学]