摘要
研究荧光定量PCR法在RNA干扰(RNA i)技术抑制宫颈癌Caski细胞人乳头瘤病毒16型E6基因(HPV16 E6)的应用情况,并与传统使用的灰度值分析法进行比较。构建shRNA重组质粒干扰载体转染至Caski细胞中。通过荧光定量PCR法和灰度值分析法检测干扰后细胞HPV16 E6基因的mRNA及蛋白质表达水平。实验表明HPV16 E6基因mRNA及蛋白质的表达量都有明显降低,与阴性对照空白质粒组相比有显著差异(P<0.01);RNA i对宫颈癌Caski细胞中HPV16 E6基因的表达具有较高的抑制率;荧光定量PCR法较之传统的灰度值分析法更为准确。
The application of fluorescent quantitative PCR in RNA interference inhibiting HPV 16 E6 gene expression of the Caski cervical cancer cell is studied and compared with densitometry analysis traditionally applied. The constructed shRNA plasmid vectors are transfected into Caski cell. The mRNA and the protein expression of HPV16 E6 gene after transfection are examined by fluorescent quantative PCR and densitometry analysis. There is a notable decrease in the mRNA expression of HPV16 E6 gene as well as the protein expression, which has siginificant difference compared with the negative control group. The result of fluorescent quantitative PCR is superior to that of densitometry analysis. RNAi can effectively inhibit the expression of HPV E6 gene in Caski cervical cancer. Fluorescent quantitative PCR, more accurate than the traditional densitometry analysis, is worthy of wide promotion in future reseaches. Quantitative analytical method with more accuracy in protein expression experiment is expected to be the breakthrough point in future reseaches.
出处
《重庆大学学报（自然科学版）》
EI
CAS
CSCD
北大核心
2007年第12期64-67,共4页
Journal of Chongqing University
基金
重庆市自然科学研究基金资助项目(CSTC2004B135124)
关键词
RNA干扰
人类乳头瘤病毒
宫颈癌
荧光定量PCR
RNA interference
HPV
cervical cancer
fluorescent quantative PCR
densitometry analysis
