高低转移肺腺癌细胞系中肿瘤抑制基因p16、p21和p53表达研究  

Expression of tumor suppressor genes p16, p21 and p53 in a pair of lung adenocarcinoma cell lines with different metastasis potentials: Anip973 and AGZY83-a

作　　者：王柏秋[1] 闫承慧[1] 吴焱[1] 高慧[1] 王琦[1] 金焰[1] 黄承滨[1] 张贵寅[1] 傅松滨[1] 李璞[1] 

机构地区：[1]哈尔滨医科大学医学遗传学教研室,150086

出　　处：《中华医学遗传学杂志》2001年第2期128-131,共4页Chinese Journal of Medical Genetics

基　　金：黑龙江省自然科学基金!(Q98- 9) ;黑龙江省科学技术计划攻关项目!(G99C2 0 - 6- 1 )&&

摘　　要：目的　探讨肿瘤抑制基因对肺腺癌细胞生长的抑制作用。方法　利用 Fu Gene转染方式分别将 p2 1和 p16基因的表达质粒转入一对肺腺癌细胞系 Anip973和 AGZY83- a中 ,同时用含野生型 p5 3基因的腺病毒感染 p16基因转染前后的这一对细胞系。对 P16和 P2 1蛋白过表达的细胞系进行了细胞生长曲线、克隆形成率、原位末端标记分析和流式细胞仪分析。结果　 p16基因的过表达只能使细胞系的 G1 期细胞比例提高 ,但细胞生长曲线、克隆形成率均未出现改变 ,未检测到凋亡信号。 P2 1蛋白过表达的一对细胞系细胞生长曲线斜率降低 ,克隆形成能力下降 ,并出现明显的 G1 期阻滞 ,但未检测到凋亡信号。p5 3基因感染 AGZY83- a、Anip973及经过 p16基因转染的细胞 AGZY83- ap16和 Anip973p16后呈现时间依赖性表达 ,细胞生长曲线和四唑盐比色法分析提示 ,野生型 p5 3基因的大量表达明显抑制以上 4种细胞的生长 ,Anip973和 Anip973p16的生长抑制率高于 AGZY83- a和 AGZY83- ap16 ;Anip973p16和 AGZY83- ap16的生长抑制率高于 Anip973和 AGZY83- a。这 4种细胞在感染 p5 3后出现典型的凋亡信号。结论　 p16基因的过表达并不能抑制细胞系的生长 ,而 p2 1基因的过表达通过 G1 期阻滞抑制这 1对肺腺癌细胞的生长 ;野生型 p5Objective: In order to investigate the suppression effect of tumor suppressor genes in lung adenocarcinoma. Methods: p16 and p21 expression vectors were transfected into a pair of lung adenocarcinoma cell lines with different metastasis potentials: Anip973 (high metastasis potential) and AGZY83-a (low metastasis potential). In the mean time, AGZY83-a, Anip973, AGZY83-ap16 and Anip973p16 were infected with recombinant adenovirus encoding wild-type p53 gene. The suppression effects of these genes were evaluated by cell growth curve, MTT, cloning efficiency assay, flow cytometric analysis and TUNEL technique. Results: Overexpression of p16 gene in Anip973 and AGZY83-a could only lengthen the G1 phase while increased expression of p21 in both of the cell lines was associated with significant lengthening of G1 phase, decreased proliferation potential and decreased cloning efficiency. High efficient expression of wild-type p53 gene in AGZY83-a, Anip973, Anip973p16 and AGZY83-ap16 inhibited the growth of these four kinds of lung cancer cells and killed the cells in the end. Apoptosis was detected in all the four kinds of cells. The suppression effect of p53 gene was higher in Anip973 and Anip973p16 than in AGZY83-a and AGZY83-ap16 while co-expression of p53 and p16 in this pair of cell lines inhibited the cells more efficiently as compared with the expression of p53 gene. Conclusion: Increased expression of p21 gene suppressed the lung adenocarcinoma cells by G1 arrest and the co-transfection of tumor suppressor genes p16 and p53 into the lung adenocarcinoma cell line proved more effective in lung cancer gene therapy.

关 键 词：高低转移肺腺癌 肿瘤抑制期 P16 P21 P53 基因表达 基因治疗 

分 类 号：R734.2[医药卫生—肿瘤] R730.54[医药卫生—临床医学]