摘要
目的探讨慢病毒介导的人乳头瘤病毒(Human papilloma virus,HPV)16型E6基因shRNA在体内对荷瘤裸鼠宫颈癌生长的抑制作用。方法将BALB/c-nu裸鼠随机分为空白对照组、干扰组和无义干扰组,经皮下接种宫颈癌Caski细胞(2×106个),移植瘤直径达0.5 cm时,空白对照组于瘤体局部注射PBS,干扰组和无义干扰组分别于瘤体局部注射靶向HPV16型E6基因的shRNA-PLL3.7干扰慢病毒和无义干扰慢病毒(3×108TU/ml),2周后检测瘤体大小和重量的变化,采用免疫组化法检测肿瘤组织中HPV16型E6、P53和P21蛋白的表达。结果与无义干扰组和空白对照组比较,干扰组裸鼠体内肿瘤明显缩小,瘤体重量明显降低(P<0.000 1),无义干扰组与空白对照组比较,差异无统计学意义(P>0.05);干扰组肿瘤组织中HPV16型E6蛋白的表达被抑制,P53和P21蛋白的表达水平明显升高。结论慢病毒介导的HPV16型E6基因shRNA能有效抑制动物体内宫颈癌的生长,具有潜在的开发应用前景。
Objective To investigate the inhibitory effect of lentivirus-mediated short hairpin RNA（shRNA） of E6 gene of human papilloma virus type 16（HPV16） on growth of cervical cancer in nude mice.Methods Nude BALB / c mice were randomly divided into blank control,interference and nonsense interference groups,and transplanted s.c.with 2 × 106 cervical cancer Caski cells.After the diameters of transplanted tumors reached 0.5 cm,PBS was injected into the tumors of mice in blank control group,while interfering lentivirus shRNA-PLL3.7 and nonsense interfering lentivirus into those in interference and nonsense interference groups respectively,each at a dosage of 3 × 10^8 TU / ml.The sizes and weights of tumors were measured 2 weeks later,in which the expressions of E6,P58 and P21 proteins of HPV16 were determined by immunohistochemical assay.Results Both the sizes and weights of tumors were significantly lower in interference group than in nonsense interference and blank control groups（P〈 0.000 1）,while showed no significant difference in the latter two groups（P 〉0.05）.The expression level of HPV16 E6 in interference group decreased,while those of P53 and P21 increased significantly.Conclusion Lentivirus-mediated HPV16 E6 shRNA inhibited the growth of cervical cancer cells in nude mice,which was potentially worthy of development and application.
出处
《中国生物制品学杂志》
CAS
CSCD
2013年第4期512-514,520,共4页
Chinese Journal of Biologicals
