雌激素对转STGC3基因CNE2细胞系生长增殖的影响  

Effect of Estrogen on CNE2 Cell Line Transfected With STGC3 Gene

作　　者：胡波[1] 邱青朝[1] 贺修胜[1] 罗桥[1] 唐国华[1] 龙治峰[1] 廖银花[1] 

机构地区：[1]南华大学肿瘤研究所,衡阳421001

出　　处：《生物化学与生物物理进展》2007年第5期538-545,共8页Progress In Biochemistry and Biophysics

基　　金：国家自然科学基金(30470967);湖南省自然科学基金(03Jjy3029);中国博士后科学基金(2004035652)资助项目~~

摘　　要：为探讨雌激素对STGC3基因抑瘤的促进作用,将重组的pcDNA3.1(+)-STGC3真核表达载体导入鼻咽癌细胞系CNE2,经G418筛选,RT-PCR及蛋白质印迹检测STGC3的表达,获得稳定高表达STGC3基因的pcDNA3.1(+)/STGC3/CNE2细胞系.采用细胞计数法,检测雌二醇(β-estradiol)对体外培养pcDNA3.1(+)/STGC3/CNE2细胞生长增殖的影响.将pcDNA3.1(+)/STGC3/CNE2细胞接种于裸小鼠前肢背部皮下,观察分析雌性与雄性裸鼠成瘤的差别.运用RT-PCR、免疫组织化学及蛋白质印迹方法,分别从mRNA及蛋白质水平,分析STGC3基因在裸鼠移植瘤组织中的表达状况.移植瘤组织病理切片检查,观察瘤细胞形态学变化.用流式细胞仪测定移植瘤组织的细胞周期分布.研究结果表明:细胞体外培养,pcDNA3.1(+)/STGC3/CNE2细胞经β-estradiol处理后,其生长速度明显减缓(P<0.05);裸鼠体内研究,接种pcDNA3.1(+)/STGC3/CNE2细胞实验组的移植瘤体积和重量均小于对照组,差异有显著性意义(P<0.05);实验组中,雌性裸鼠组移植瘤体积和重量均小于雄性组,差异有显著性意义(P<0.05),雌性裸鼠组移植瘤生长最慢,而对照组中雄性与雌性裸鼠组间瘤块的差异无显著性意义(P>0.05);接种pcDNA3.1(+)/STGC3/CNE2细胞的雌性裸鼠组移植瘤,阻滞于G0/G1期细胞数大于其他各组(P<0.05).上述体内外研究结果显示,雌激素可能具有增强STGC3基因对CNE2细胞系的生长抑制作用.To investigate the effect of estrogen to STGC3 gene on CNE2 cell line in vitro and in vivo. The recombinant pcDNA3.1 （＋）/STGC3 plasmid was constructed and transfected into CNE2 cell line with lipofectamine 2000. CNE2 cell clones with stable STGC3 expression were obtained through G418 selection and identified by RT-PCR and Western blotting methods. The effect of β-estradiol on growth rate of pcDNA3.1 （＋）/ STGC3/CNE2 cell line was examined by cytometry. The pcDNA3.1 （＋）/STGC3/CNE2 cell line was inoculated subcutaneously in nude mice. Tumorigenicity diversity was analyzed in female and male nude mice. STGC3 expression was detected in tumor xenografts in nude mice by RT-PCR, immunohistochemistry and Western blotting methods. Cell morphologic changes were observed by microscope. Flow cytometry was used to analyze cell cycles. The results indicated that pcDNA3.1 （＋）/STGC3/CNE2 cell growth rate decreased after treated with β-estradiol in vitro. There were significant differences in tumor size and mass between pcDNA3.1 （＋）/ STGC3/CNE2 and control cases （P 〈 0.05）. Furthermore, tumor size and mass had significant differences between male and female nude mice in the STGC3 transfection cases and tumor growth rate of pcDNA3.1 （＋）/ STGC3/CNE2 in the female nude mice was the lowest in all cases （P 〈 0.05）. No significant difference was found between male and female nude mice in other control cases （P 〉 0.05）. There were more cells blocked in G0/GI phase in tumor xenografts ofpcDNA3.1 （＋）/STGC3/CNE2 cell line of the female nude mice case than that in other cases. Data above indicated that estrogen might enhance the effect of STGC3 to inhibit CNE2 cell proliferation in vitro and in vivo.

关 键 词：STGC3基因 雌激素 鼻咽癌 CNE2细胞系 裸鼠 

分 类 号：R739.63[医药卫生—肿瘤]