α1，2-岩藻糖转移酶基因转染对卵巢癌细胞生物学行为的影响  

α1,2-FUCOSYLTRANSFERASE GENE TRANSFECTION INFLUENCES ON BIOLOGICAL BEHAVIOR OF OVARIAN CARCINOMA-DERIVED RMG-I CELLS

作　　者：郝莹莹[1] 林蓓[1] 赵越[1] 张宇华[2] 李飞飞[1] 刁斌[3] 欧阳玲[1] 张淑兰[1] 

机构地区：[1]中国医科大学附属盛京医院妇产科,沈阳110004 [2]辽宁省肿瘤医院,沈阳110042 [3]沈阳市第四医院,沈阳110031

出　　处：《分子细胞生物学报》2008年第6期435-442,共8页Journal of Molecular Cell Biology

基　　金：国家自然基金资助项目(30170980;30571958);归国人员博士启动基金项目(2000479);辽宁省博士启动基金项目(001040)

摘　　要：为探讨α1,2-岩藻糖转移酶(α1,2-fucosvltransfemse,α1,2-FT)基因转染对卵巢癌细胞RMG-(?)生物学特性的影响,利用PCR方法克隆人α1,2-FT基因的编码区HFUT-H,构建表达载体pcDNA3.1-HFUT-H,利用磷酸钙法将其转染入卵巢癌细胞系RMG-(?),建立α1,2-FT基因稳定高表达细胞株RMG-I-H。采用RT-PCR及α1,2-FT活性测定检测转染前后细胞系α1.2-FT基因表达及α1,2-FT活性的变化,利用免疫细胞化学方法测定细胞表面Lewis y抗原表达变化.利用MTT法、流式细胞仪细胞增殖周期检测方法测定转染前后细胞系生物学特性的变化。分别以转染前细胞RMG-I和转染空载体细胞RMG-I-C为对照组。结果显示转染后细胞α1,2-FT mRNA表达增高,α1,2-FT活性增加20-30倍。转染后的细胞易复层生长,生长速度加快。细胞克隆边缘呈树突状延伸生长。转染后细胞RMG-I-H软琼脂克隆形成率(35%)明显高于对照组RMG-I (13%)及RMG-I-C(12%),为对照组的2.6倍。RMG-I-H细胞G_1期比例从对照组的74.14%下降到59.46%,G_2-M期和S期比例分别从对照组的2.05%和23.95%上升到7.32%和33.27%(P< 0.05)。这些结果充分说明,α1,2-FT及Lewis y抗原具有促进RMG-I细胞增殖,提高RMG-I细胞生存能力,促进肿瘤发生、发展的作用。To explore the effect of α1,2-fucosyltransferase （α1,2-FT） gene on the biological behavior of ovarian carcinoma cells, the expression vector pcDNA3.1-HFUT-H which containing human α1,2-FT gene coding region was constructed and transfected into ovarian carcinoma cell line RMG-I. α1,2-FF gene expression, Lewis y antigen expression, α1,2-FT activity, rates of cell growth and tumorogenesis in nude mice were compared before and after RMG-I being transfected. The transfection of cxl, 2-FT gene significantly increased the expression of α1,2-FT gene and Lewis y antigen in RMG-I cells. After the transfection, RMG-I cells showed remarkable improvement of α1,2-FT activity, cell proliferation, the ability of colony formation in soft agar and tumorogenesis in nude mice. The growth rates of α1,2-FF gene transfected RMG-I cells were obviously increased, presenting that the proportion of the cells in G2-M and S phases was obviously increased from 2.05% to 7.32% and from 23.95% to 33.27% respectively, and that of the cells in G1 phase was obviously decreased from 74.14% to 59.46%, as compared with the control group. These findings indicate that α1,2-FF and Lewis y antigen have the ability to enhance the proliferation and elevate the survival rates of RMG-I cells, which can promote the genesis and development of ovarian carcinoma.

关 键 词：α1 2-岩藻糖转移酶基因 转染Lewis y抗原 卵巢癌 

分 类 号：R737.31[医药卫生—肿瘤]