ODC反义RNA表达质粒的构建及其对人肺鳞癌细胞生长特性的影响  

Construction of ODC Antisense RNA Expression Piasmid and Its Effect on the Growth Characteristics of Human Lung Squamous Carcinoma Cells

作　　者：关钧[1] 范慕贞[1] 曹淑兰[1] 

机构地区：[1]中日友好临床医学研究所生化及分子生物学研究室,北京100029

出　　处：《中国肿瘤生物治疗杂志》1995年第1期21-24,共4页Chinese Journal of Cancer Biotherapy

基　　金：国家自然科学基金资助项目(39470793)

摘　　要：鸟氨酸脱羧酶(ODC)是多胺生物合成途径中的限速酶,其活性的异常升高及多胺堆积与恶性肿瘤的发生、发展及转移密切相关.作者构建一个能表达人ODC反义RNA的真核表达质粒pCMV/ODCas,用磷酸钙沉淀法将其导入人肺鳞癌细胞系LTEP-78(L78),经G418筛选,获两株转染细胞系L78/ODCasl和L78/ODCas4.与亲本细胞相比,其生长速度减慢;软琼脂集落形成能力减弱,DNA、RNA及蛋白质等生物合成受到抑制,ODC酶活性降低.经Southern blot及PCR分析,进一步证明了上述变化与ODC反义序列稳定完整地整合到转染细胞基因组中有关.Orinthine decarboxylase (ODC) is a rate-limiting enzyme in polyamine biosynthesis. Abnormal elevation of ODC activity and subsequent polyamine accumulation are intimately associated with the genesis, development and metastasis of cancer. ODC antisense RNA expression plasmid, pCMV/ODCas, was constructed, with which the cells of human lung squamous carcinoma were transfected by the calcium phosphate precipitation method. After G418 selection, two G418 resistant colonies were obtained. The growth rate, the ability to form colony in soft agar, the biosyntheses of DNA, RNA and proteins, and ODC activities of the two antisense transfectants were significantly suppressed in comparison with the parental cell line. It was demonstrated that these changes were associated with the stable integration of exogenous antisense ODC gene in the genomic DNA of transfectants by analysis of Southern blot and PCR.

关 键 词：ODC 肺鳞癌细胞 反义RNA 表达质粒 转染细胞 多胺生物合成 集落形成 基因组 DNA 磷酸钙沉淀法 

分 类 号：R734.2[医药卫生—肿瘤] R914.2[医药卫生—临床医学]