bax和p53基因共转染对人舌癌细胞增殖和凋亡的影晌

Effect on Proliferation and Apoptosis of Human Lingual Carc inoma Cells Cotransfected by bax and p53 Genes

目的 探讨诱导凋亡的 bax基因和 p5 3抑癌基因共转染对人癌细胞增殖和凋亡的作用。方法 构建 p SV- CIP- bax- CAT嵌合基因。在其 bax基因上游含人 a1( )型胶原基因的第一内含子 2 17bp片段 ( + 82 2～ +10 93)和 317bp启动子片段。经阳离子转染试剂 DOSPER介导 ,分别用 p SV- CIP- bax- CAT转染、p SV- CIP- bax-CAT和 p SV- p5 3- CAT共转染培养的人舌癌 Tca8113细胞系 ( L CC)。结果 免疫狭缝印迹和 EL ISA检测证实 ,转染组和共转染组比对照的 bax和 p5 3基因表达量明显增加。 MTT显色分析、TU NEL荧光显微镜和流式细胞仪监测结果显示 ,bax基因或 p5 3基因均具抑制 L CC增殖和诱导凋亡的作用 ( bax组和 bax+ p5 3组的抑制率分别为2 3.9%和 2 6 .1% )。结论 人 a1( )型胶原基因的顺式作用元件能促使 bax基因在 L CC异位表达 ;bax与 p5 3共转染 48小时 ,对 L

Objective To investigate th e effects on prolife ration and apoptosis of human cancerous cells cotransfected by bax apoptosis in ducing gene and p53 tumor suppressor gene. Methods The chi meric gene pSV CIP bax CAT was constructed in which bax gene was flanked upstream by a 217bp fragment (+822^+1093) of the first intron and a 317bp promoter fragment of human a 1(Ⅰ) colla gen gene . Human lingual carcinoma cell line Tca 8113 (LCC) in culture was respe ctively transfected with pSV CIP bax CAT, and cotransfected with pSV p53 CA T and pSV CIP bax CAT by using the transfection reagent DOSPER. Res ults Immuno slot blot and ELISA demonstrated that the expression of bax and p53 genes incr eased remarkably in the transfected and cotransfected LCC, compared with the con trols(LCC transfected with pSV CIP CAT and LCC).MTT colorimetric assay,TUNE L fluorecence microscopy and flow cytometry showed that foreign bax or p53 gene inhibited the LCC growth and induced apoptosis(23.9% and 26.1% of inhibitory ra te by bax gene and p53 plus bax genes respectively). Conclusion The ectopic expression of the bax gene in the LCC promoted by the cis act ing elements of human a1(Ⅰ)collagen gene has obviously synergistic effect on th e proliferation and apoptosis of the LCC cotransfected with p53 gene plus bax gene for 48 hours.