Survivin shRNA-APC双基因共表达稳转株对人HT-29结肠癌细胞中PCNA、Ki-67表达的影响  

Effects of Survivin shRNA-APC co-expression stable transfected strain on the expression of PCNA and Ki-67 in human HT-29 colon cancer cells

作　　者：袁喜先 张蒙蒙 曲若梅[1] 张玉健 孙元佳 陈月 张书娟 袁晓岚 YUAN Xi-xian;ZHANG Meng-meng;QU Ruo-mei;ZHANG Yu-jian;SUN Yuan-jia;CHEN Yue;ZHANG Shu-juan;YUAN Xiao-lan(First Affiliated Hospital of Jiamusi University,Jiamusi,Heilongjiang 154003,China)

机构地区：[1]佳木斯大学附属第一医院,黑龙江佳木斯154003 [2]佳木斯大学临床医学院,黑龙江佳木斯154007

出　　处：《中国临床研究》2019年第5期597-599,603,共4页Chinese Journal of Clinical Research

摘　　要：目的探讨生存素短发夹RNA-腺瘤性结肠息肉病(SurvivinshRNA-APC)双基因共表达稳转株对HT-29结肠癌细胞裸鼠皮下移植瘤组织细胞中增殖细胞核抗原(PCNA)、Ki-67表达的影响。方法将30只SPF级裸鼠随机分为5组,为SurvivinshRNA-APC双基因组、SurvivinshRNA及APC单基因组、空载组和阴性对照组,对各组稳转株及HT-29结肠癌细胞分别进行培养,PBS重悬使癌细胞浓度达到2×10^6/ml,于每只裸鼠右前腋下分别接种相对应的细胞悬液,构建移植瘤模型,每6天测量移植瘤体积,待7周后统一处死所有裸鼠,剥离移植瘤,将瘤体组织用多聚甲醛固定,石蜡包埋切片,根据免疫组化检测人HT-29结肠癌细胞裸鼠皮下移植瘤组织细胞中PCNA、Ki-67表达。结果免疫组化结果显示,Ki-67蛋白及PCNA蛋白主要定位于移植瘤组织细胞的胞核内;在阴性对照组和空载组均可见较多棕黄色或棕褐色颗粒,双基因组细胞内可见极少量黄色或淡黄色细颗粒,APC组及SurvivinshRNA组染色程度介于二者之间。APC组、SurvivinshRNA组、双基因组PCNA、Ki-67蛋白表达积分分别低于阴性对照组及空载组(P<0.01);双基因组PCNA、Ki-67蛋白表达积分分别低于APC组、SurvivinshRNA组(P<0.01);阴性对照组与空载组比较、APC组与SurvivinshRNA组比较,PCNA、Ki-67蛋白表达积分差异无统计学意义(P>0.05)。结论SurvivinshRNA-APC双基因共表达稳转株可通过下调HT-29结肠癌细胞裸鼠皮下移植瘤组织细胞中PCNA、Ki-67的表达,抑制移植瘤细胞增殖,其效果较单基因稳转株更为显著。Objective To investigate the effect Survivin shRNA-APC co-expression stable transfected strain on the expression of PCNA and Ki-67 in HT-29 colon cancer xenograft cells in nude mice.Methods Thirty SPF-grade nude mice were randomly divided into five groups: Survivin shRNA-APC double genome group, Survivin shRNA group, APC group, blank group and control group.Steady transfected lines and HT-29 colon cancer cells were cultured in five groups, respectively.Resuspend cancer cells in PBS to achieve a concentration of 2 × 10^6 /ml.Cell suspension was inoculated into the right anterior axilla of each nude mouse to construct the transplanted tumor model.The volume of transplanted tumors was measured every 6 days.After 7 weeks,all nude mice were executed.The transplanted tumors were peeled off and fixed with paraformaldehyde.The expression of PCNA and Ki-67 in human HT-29 colon cancer cells was detected by immunohistochemistry.Results Immunohistochemical results showed that Ki-67 protein and PCNA protein were mainly located in the nucleus of transplanted tumor cells;brown or brown granules could be seen in both control group and blank group;a small amount of yellow or light yellow fine granules could be seen in survivin shRNA-APC double genome group;the staining degree of APC group and Survivin shRNA group was between the two groups.The expression of PCNA and Ki- 67 protein increased in the order of double genome group→APC group, Survivin shRNA group→blank group and control group ( P < 0.01).Conclusion Survivin shRNA-APC co-expressing stable transfection could significantly inhibit the proliferation of transplanted tumor cells by down-regulating the expression of PCNA and Ki-67 in HT-29 colon cancer cells subcutaneously transplanted in nude mice,and its effect is more significant than that of single gene stable transfection.

关 键 词：结肠癌 增殖 增殖细胞核抗原 Ki-67 生存素 生存素短发夹RNA 腺瘤性结肠息肉病 

分 类 号：R735.35[医药卫生—肿瘤]