论文部分内容阅读
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 CC抑制增殖和诱导凋亡的作用具明显协同效应
Objective To investigate the effect of co-transfection of apoptosis-inducing bax gene and p5 3 tumor suppressor gene on proliferation and apoptosis of human cancer cells. Methods The p SV-CIP-bax-CAT chimeric gene was constructed. The first intron 2 17 bp fragment (+82 2 to +10 93) and the 317 bp promoter fragment containing the human a1 (type) collagen gene were upstream of their bax genes. The human tongue cancer Tca8113 cell line (cotransfected with p SV-CIP-bax-CAT, p SV-CIP-bax-CAT and p SV-p5 3- L CC). Results The results of immunostaining and ELISA showed that the expression of bax and p5 3 genes in transfected and co-transfected cells were significantly increased compared with the control. MTT colorimetric analysis, TU NEL fluorescence microscopy and flow cytometry monitoring results showed that either bax gene or p5 3 gene inhibited L CC proliferation and induced apoptosis (bax group and bax + p5 3 groups were 2 3.9% and 26.1%). Conclusions Cis-acting elements of human a1 (type) collagen gene can promote ectopic expression of bax gene in L CC; cotransfection of bax with p5 3 for 48 hours has obvious synergistic effect on L CC inhibition and apoptosis induction