两株肿瘤细胞系HLAI类分子基因表达水平鉴定及意义.docVIP

两株肿瘤细胞系HLAI类分子基因表达水平鉴定及意义 　 作者：关德莹，沈晗，邵红伟，黄树林 【摘要】 目的 探讨两株常见肿瘤细胞系Hela细胞和BEL7402细胞HLAI类分子基因表达水平及其 影响 CTL反应性的可能意义。 方法 提取肿瘤细胞的基因组DNA，利用PCR反应在基因组水平鉴定HLAI类分子基因的拷贝数情况；提取细胞总RNA， RTPCR反应合成cDNA，利用PCR反应鉴定肿瘤细胞HLAI类分子在RNA水平的表达情况。结果 两株肿瘤细胞的HLAI类基因在基因组水平的拷贝数无明显下降；在RNA水平上，Hela细胞的HLAA、HLAB和HLAC基因均有表达，表达水平无明显下降，BEL7402细胞的HLAA和HLAC基因有表达，但HLAC基因表达水平降低，而HLAB基因未见明显表达。 结论 RNA水平上，部分HLAI类分子基因表达的降低或缺失推测可能是导致BEL7402细胞比Hela细胞出现CTL反应性下降的原因之一。 论文代写 【关键词】 HLAI类分子；Hela细胞；BEL7402细胞；mRNA 　　Abstract:Objective To investigate the expression level of HLAI molecule genes in two tumor cell lines, Hela and BEL7402 and the possible significance of CTL reactions.Methods Genome DNA were extracted from these two tumor lines for detecting the HLAI molecule genes′ levels by PCR.Total RNA from two cell lines were extracted for RTPCR, HLAI molecule genes′expression on RNA level were judged by PCR and agarose gel electrophoresis.Results HLAI molecule genes levels did not decrease significantly on genome level.On RNA level, HLAA, HLAB and HLAC all expressed normally without significant descent in Hela cell line. The expression of HLAA and HLAC were both detected in BEL7402 cell line, but the expression of HLAC gene showed significant decease, and no expression of HLAB gene were detected on RNA level.Conclusion At RNA level, reduction or absence HLAI molecule genes′ expression of Hela cell line were not found, however, the partial descent and absence of HLAI molecule genes′ expression in BEL7402 cell line was the possible reason for the reduction of CTL reaction. 毕业论文 　　Key words:HLAI molecule；Hela cell line ；BEL7402 cell line；mRNA 毕业论文 　　人类主要组织相容性复合物（MHC），又称为人类白细胞抗原(human leukocyte antigen，HLA )系统，位于第6号染色体短臂（6p21.31）约3.6Mb［1］，按功能差别可大致分为HLAI类分子和HLAII类分子两大类，其中HLAI类分子在机体的细胞免疫中发挥重要作用。 研究 发现，抗肿瘤免疫的核心是机体针对肿瘤的细胞免疫反应，其中CD8+CTL细胞必需依靠肿瘤自身HLAI类分子呈递的肿瘤抗原才能得以活化发挥杀伤作用。但由于肿瘤细胞的遗传不稳定性，会导致HLAI类分子的表达下降或缺失，是引起肿瘤免疫耐受形成的重要原因［2, 3］。因此，了解肿瘤细胞的HLAI类分子表达情况对研究如何打破免疫耐受，开展过继性细胞学