pGPU6/GFP/Neo-hTERT-shRNA对结直肠癌SW480细胞裸鼠移植瘤的抑制  

Inhibition of pGPU6/GFP/Neo-hTERT-shRNA on colorectal cancer SW480 cell xenograft in nude mice

作　　者：蔡艳玲[1] 罗小玲[1] 葛连英[2] 刘爱群[2] 谢裕安[1] 

机构地区：[1]广西医科大学附属肿瘤医院生物治疗科,广西壮族自治区南宁530021 [2]广西医科大学附属肿瘤医院内镜室,广西壮族自治区南宁530021

出　　处：《中国肿瘤生物治疗杂志》2012年第1期72-76,共5页Chinese Journal of Cancer Biotherapy

基　　金：广西自然科学基金资助项目(No.2010GXNSFA013238);广西卫生厅重点课题资助项目(No.Z200971)~~

摘　　要：目的:研究靶向hTERT基因的重组质粒pGPU6/GFP/Neo-hTERT-shRNA对人结直肠癌SW480细胞裸鼠移植瘤的治疗作用。方法:于裸鼠右侧腋下皮下注射人结直肠癌SW480细胞建立结直肠癌移植瘤动物模型,随机分为生理盐水组(NS组)、pGPU6/GFP/Neo-NC-shRNA组(NC-shRNA组)和pGPU6/GFP/Neo-hTERT-shRNA组(hTERT-shRNA组),各组连续进行相应治疗6次后,观察肿瘤的生长状况,测量肿瘤体积并绘制肿瘤生长曲线,H-E染色观察肿瘤组织形态学变化,免疫组织化学法检测移植瘤组织中hTERT蛋白的表达,TUNEL法检肿瘤组织中细胞凋亡情况,RT-PCR法检测瘤组织中hTERT mR-NA的表达。结果:与NC-shRNA组和NS组比较,hTERT-shRNA组移植瘤体积增长速度减慢;hTERT-shRNA组移植瘤组织中见肿瘤细胞形态明显改变,凋亡细胞数明显增多[(36.30±5.05)%vs(5.25±1.06)%、(6.95±1.07)%,P<0.01];hTERT-shRNA组hTERT的mRNA和蛋白表达均明显受到抑制(171.42±30.94 vs 146.89±21.43、137.35±25.49,P<0.01;0.39±0.09 vs 0.81±0.335、0.750±0.206,P<0.05)。结论:重组质粒pGPU6/GFP/Neo-hTERT-shRNA通过下调hTERT mRNA和蛋白水平的表达促进肿瘤细胞的凋亡,抑制结直肠癌移植瘤的生长。Objective：To investigate the treatment effect of recombinant plasmid pGPU6/GFP/Neo-hTERT-shRNA targeting hTERT gene on human colorectal cancer SW480 cell xenograft in nude mice.Methods： Human colorectal cancer SW480 cells were subcutaneously implanted under the skin of the right armpit to establish nude mice model of colorectal cancer,after the tumors grew to a definite size.The mice were randomly divided into three groups： normal saline（NS group）,pGPU6/GFP/Neo-NC-shRNA group（NC-shRNA group）and pGPU6/GFP/Neo-hTERT-shRNA group（hTERT-shRNA group）.After each group was treated for 6 consecutive times,the growth status of the tumor was observed,tumor volume was measured,tumor growth curve was drawn,tumor tissue morphology was observed with H-E staining,the expression of hTERT protein in the tumors was detected by immunohistochemistry,cell apoptosis was inspected by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling（TUNEL）,and the expression of hTERT mRNA was checked by RT-PCR.Results： The growth of tumor volume became slower in hTERT-shRNA group than did that in NS group and NC-shRNA group.Compared with NS group and NC-shRNA group,the tumor cell morphology changed obviously and the number of apoptotic cells increased significantly in the transplanted tumor tissues in hTERT-shRNA group（[36.3±5.05]% vs [5.25±1.06]%,[6.95±1.07]%,P0.01）.Compared with NS group and NC-shRNA group,the expression of hTERT protein was significantly inhibited in hTERT-shRNA group（[171.42±30.94] vs [146.89±21.43],[137.35±25.49],P0.01）.Compared with NS group and NC-shRNA group,the expression of hTERT mRNA was significantly inhibited in hTERT-shRNA group（0.39±0.09 vs 0.81±0.34,0.75±0.21,P0.05）.Conclusion： Recombinant plasmid pGPU6/GFP/Neo-hTERT-shRNA promotes apoptosis of implanted human colorectal cancer by down-regulating the expression of hTERT mRNA and hTERT protein in tumor tissues,thus inhibiting the growth.

关 键 词：RNA干扰 HTERT基因 结直肠癌 裸鼠 凋亡 

分 类 号：R735.3[医药卫生—肿瘤] R730.54[医药卫生—临床医学]