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目的 观察HepG2细胞因乙型肝炎病毒X抗原 (HBx)转染导致的基因表达差异。 方法①用逆转录病毒感染的方法建立表达乙型肝炎病毒X抗原 (HepG2X)及氯霉素乙酰转移酶 (HepG2Cat)的细胞模型。②用抑制差减杂交的方法做HepG2X和对照细胞 (HepG2Cat)的cDNA文库差示筛选。③用3 2 P随机引物标记法标记探针做NorthernBlot杂交及地高辛缺口翻译标记法做原位分子杂交以筛选基因探针。结果 经cDNA文库差减杂交及抑制性PCR扩增共得到 10个cDNA探针 ,其中 8个被HBx所启动 ,2个因HBx而表达下降。经检测 ,这些基因在HepG2X和对照细胞中的表达均有差异。在被HBx所抑制的 2个差示表达基因中 ,1个与人的蛋白翻译起始因子同源 ,同时发现 ,此基因在正常组织中表达较强 ,而在肿瘤组织中表达被抑制。结论 HBx能够改变宿主细胞的基因表达 ,并且这些基因变化可能和肝癌发生有关
Objective To observe the difference of gene expression in HepG2 cells caused by hepatitis B virus X antigen (HBx) transfection. Method 1 A cell model expressing hepatitis B virus X antigen (HepG2X) and chloramphenicol acetyltransferase (HepG2Cat) was established by retroviral infection. 2 HepG2X and control cells (HepG2Cat) were differentially screened by inhibition subtractive hybridization. 3 Use the 3 2 P random primers to label the probes for Northern Blot hybridization and digoxin gap translation labeling for in situ molecular hybridization to screen for gene probes. Results A total of 10 cDNA probes were obtained by cDNA library subtractive hybridization and suppression PCR amplification, of which 8 were initiated by HBx and 2 were decreased by HBx. Upon detection, the expression of these genes was different in HepG2X and control cells. Of the two differentially expressed genes that were inhibited by HBx, one was homologous to human protein translation initiation factor. It was also found that this gene is strongly expressed in normal tissues and suppressed in tumor tissues. Conclusion HBx can alter the gene expression of host cells, and these gene changes may be related to the occurrence of liver cancer.