人肺癌细胞MCT2基因的克隆及其mRNA表达分析

目的　克隆人肺癌细胞单羧酸转运蛋白第二亚型基因 (MonocarboxylateTransporter 2 ,MCT2 )的编码序列 ,并对其在肿瘤组织细胞mRNA表达作半定量分析。方法　 (1)通过RT PCR方法克隆人肺腺癌A5 4 9细胞中的MCT2基因片段。应用四色荧光、双脱氧终止法测序 ;通过Genebank数据库应用分析软件对克隆基因序列进行同源性分析。 (2 )采用半定量RT PCR方法比较人肺癌组织、癌旁正常肺组织及人肺癌细胞株中MCT2mRNA的表达水平。结果　 (1)应用RT PCR方法克隆出人肺癌细胞的MCT2基因片段 ;肺癌细胞此基因编码序列与人肝细胞MCT2基因的cDNA编码序列具有高度同源性。 (2 )人肺癌组织及人肺癌细胞株中MCT2mRNA的表达非常显著地高于癌旁正常肺组织 (P <0 .0 0 1)。结论　MCT2基因为一序列保守的基因 ,在肿瘤细胞中超表达并可能对细胞能量代谢及生长增殖具有重要作用

Cloning monocarboxylate transporter 2 gene of human lung cancer cells and analyzing its mRNA expression level

Objective 1.To clone and analyze the homologous character of monocarboxylate transporter 2 (MCT2)gene code sequence in human lung cancer cells. 2. To detect the expression level of MCT2 mRNA in human lung cancer cells.Methods MCT2 code sequence of human lung cancer A549 cells was cloned by RT PCR method and sequenced with four fluorescence color test and Sanger's method ;The homologous features of the sequence was analyzed with Genebank database by software. Using the semi quantative RT PCR method, the content of MCT2 mRNA in human lung cancer cells and normal cells was examined . Results (1)MCT2 gene of human pulmonary adenocarcinoma cells was cloned and the squence of the gene is high homologous with the code region of human hepatic cells MCT2. (2) The expression level of MCT2 mRNA in human lung cancer cells is very significantly high, compare to lung normal cells( P <0.001). Conclusion The sequence of MCT2 gene is conservative. The up regulation of MCT2 mRNA in human lung cancer cells might play an important role to their energy metablism and growth .