摘要
目的观察人β-防御素1(hβD1)对宫颈癌Hela细胞HPV18复制和表达的影响。方法基因转染:通过FugenHD介导,将已构建的hβD1/psectag质粒转染至Hela细胞(实验组),并设空载组和空白组。通过免疫细胞化学染色观察转染后目的基因在细胞中的表达;荧光定量PCR检测转染48 h和72 h后Hela细胞HPV18DNA拷贝数的变化;半定量RT-PCR检测转染后48 h和72 h的Hela细胞HPV18E6 mRNA表达的改变。结果转染hβD1/psectag质粒48 h和72 h后的Hela细胞均有hβD1的表达,后者明显增强,而空载组和空白组未见表达。较之空载组和空白组,实验组48 h HPV18 DNA拷贝数增多,72 h HPV18 DNA拷贝数减少,但差异无统计学意义。实验组48 h的Hela细胞HPV18E6 mRNA表达较空载组和空白组未见明显改变,而72 h的Hela细胞HPV18E6 mRNA表达较空白组减少(P<0.05)。结论hβD1对Hela细胞HPV18E6 mRNA表达有抑制作用并呈浓度依赖性,而对其DNA复制作用不明显。
Objective Investigate the influences of human β-defensinl （hβD1） on the replication and expression of HPV18 in Hela cell. Methods Gene transfection., mediated by Fugen HD, hβD1/psectag plasmid was transfected to Hela cell [hβD1/psectag ： liposome complexes（hβD1/psectag z lip）group], and control groups [psectag：liposome complexes（psectag ： lip）group and blank group] were also established. After transfection, the expression of the target gene in Hela cell was investigated by the method of immunocytochemistry. 48 h and 72 h after the transfection, the change of the copy number of HPV18 DNA in Hela cell was investigated respectively by the quantitative fluorescent PCR method, and the change of the HPV18 E6 mRNA in Hela cell was evaluated by the semiquantitative RT-PCR. Results 48 h and 72 h after the transfection of hβD1/psectag plasmid to Hela cell, hβD1 was expressed in both of the two groups, and the latter showed a tendency of stronger expression. Compared with the control groups, the copy number of HPV18 DNA in Hela cell in the hβD1/psectag ： lip group increased at 48 h and decreased at 72 h after the transfection, but the change was not statistically significant. 48 h after the transfection, compared with the control groups, the expression of HPV18 E6 mRNA in Hela cell in the hβD1/ psectag ： lip group changed a little; and 72 h after the transfection, the expression level of HPV18 E6 mRNA decreased significantly （P〈0.05）. Conclusion hβD1 displayed an inhibitory effect to the expression of HPV18 mRNA in Hela cell in a concentration-dependent pattern, but no significant effect on the duplication of HPV18 DNA.
出处
《四川大学学报（医学版）》
CAS
CSCD
北大核心
2009年第2期232-235,共4页
Journal of Sichuan University(Medical Sciences)
关键词
人β-防御素1
人乳头瘤病毒
宫颈肿瘤
基因表达
基因复制
Human β-defensinl （hβD1） Human papilloma virus Cervix uteri tumor Gene expression Gene duplication
