单羧酸转运蛋白反义基因转染人肺腺癌细胞对其生物学特性的影响  

Effect of antisense transfection of monocarboxylate transporter on cell biological characteristics in human lung adenocarcinoma cells

作　　者：张桂芝[1] 黄桂君[1] 郭先健[1] 钱桂生[1] 

机构地区：[1]第三军医大学附属新桥医院全军呼吸内科研究所,重庆400037

出　　处：《第三军医大学学报》2001年第2期128-130,共3页Journal of Third Military Medical University

基　　金：国家自然科学基金资助项目 !( 3990 0 0 6 7);重庆市科委课题

摘　　要：目的　研究单羧酸转运蛋白基因第一亚型 (MCT1)反义表达载体转染肿瘤细胞对其细胞内pH(pHi)调节、乳酸转运及生长性质的影响。方法　通过电穿孔法将单羧酸转运蛋白MCT1基因反义表达重组子pLXSN MCT1转染于肺腺癌细胞系A5 49中 ,经G418筛选转染细胞阳性克隆。用PCR方法鉴定MCT1反义表达重组载体在基因组的转染整合 ;以分光光度法测定pHi及乳酸含量变化 ,并以细胞生长曲线研究细胞的生长情况。结果　与未转染的A5 49细胞比较 ,转染MCT1反义表达重组载体的细胞pHi降低、乳酸显著升高 (P <0 .0 0 1) ;且细胞的生长受到明显抑制。结论　单羧酸转运蛋白MCT1基因在肿瘤细胞pHi调节、乳酸转运及细胞的生长中起着重要的调节作用。Objective To study the effect of transfecting anti sense expression vector of the first subtype of the monocarboxylate transporter (MCT1) gene into human lung adenocarcinoma cells on intracellular pH (pHi) regulation, lactate transportation and cell growth. Methods MCT1 antisense gene recombinant vector pLXSN MCT1 was introduced into human lung cancer cells A549 with electroporation. The cell colonies resistant to G418 were selected. Positive clones were examined by PCR to confirm the integration of genomic A549 DNA and antisene MCT1 gene. The changes of pHi and lactate transportation were detected with spectrophotometry. Cell growth was studied with cell growth curve. Results pHi and lactate transport were remarkably decreased in the transfected cells, and the cell growth was inhibited compared with the cells without transfection( P < 0.001) . Conclusion MCT1 gene may play an important role in pHi regulation, lactate transport and cell growth in lung tumor cells.

关 键 词：单羧酸转运蛋白基因 基因转染 PHI 乳酸 肺腺瘤 

分 类 号：R734.2[医药卫生—肿瘤]