HPV16编码蛋白与宫颈癌细胞内CALCA和TFPI-2表达的关系

doi:10.3969/j.issn.1004-616x.2017.06.006

Abstract

Abstract: OBJECTIVE:We investigated the expression of CALCA and TFPI-2,and their association with expression of human papillomaviurs (HPV) 16 coding proteins in cervical carcinoma cells. METHODS:We designed and synthesized small-hairpin RNAs (shRNA) which were specific to the sequences of E6 or E7 genes of HPV16. In addition,we prepared a series of shRNA expression constructs using pRNAi plasmid that could simultaneously express shRNA and green fluorescent protein (GFP) reporter gene. After transient expression of shRNA vectors in HPV16-positive SiHa cervical carcinoma cells by lipid transfection,we analyzed transfection efficiency,inhibition of HPV-coding gene expression and its effect on candidate gene expression using fluorescent imaging and quantitative RT-PCR. RESULTS:Relatively high transfection efficiency was confirmed by using confocal microscopy on cell population based on their release of green fluorescence. According to quantitative RT-PCR analysis,E6 or E7 expressions were,to a very high extent,inhibited after transfection of shRNA vectors. Expressions of CDK4 and BCL-2 were significantly decreased after selective inhibition of E6 or E7 gene expression. These observations suggest that the inhibition of HPV16 coding gene expression might have inhibited cell proliferation and induced cell-cycle arrest. Expressions of CALCA and TFPI-2 were significantly upregulated after inhibition of E7 gene expression but not remarkable after inhibition of E6 gene expression. CONCLUSION:Results of this study together with previous reports suggest that HPV16 E7 protein may promote the downregulation of CALCA and TFPI-2 in cervical carcinoma cells. The mechanism may be involved with HPV-driven cervical carcinogenesis.

Key words: HPV16, coding proteins, SiHa cells, cervical carcinoma, CALCA, TFPI-2

CLC Number:

R349.7

DILARE Lidifu, MUTALIFU Aimaiti, SHENG Lei, GUZHALINUER Maitisha, AIERKEN Rozibilali, ABULIZI Abudula. Expression of HPV16 coding proteins and regulation of CALCA and TFPI-2 expression in cervical carcinoma cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2017, 29(6): 431-436.

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Expression of HPV16 coding proteins and regulation of CALCA and TFPI-2 expression in cervical carcinoma cells

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