The effect of mutations in the upstream regulatory region of HPV-16 on the activity of virus early promoter

宋　丹; 史　茜; 侯向前

doi:10.19401/j.cnki.1007-3639.2017.02.005

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The effect of mutations in the upstream regulatory region of HPV-16 on the activity of virus early promoter

更新时间：2025-12-31

- The effect of mutations in the upstream regulatory region of HPV-16 on the activity of virus early promoter
- China Oncology Vol. 27, Issue 2, Pages: 109-114(2017)
- 作者机构：
  
  新疆大学生命科学与技术学院,新疆,乌鲁木齐,830046
- 作者简介：
- 基金信息：
- DOI：10.19401/j.cnki.1007-3639.2017.02.005
  CLC：
- Published Online：22 March 2017，
  
  Published：22 March 2017
- 稿件说明：
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宋　丹,史　茜,侯向前,等. HPV-16

宋　丹, 史　茜, 侯向前. The effect of mutations in the upstream regulatory region of HPV-16 on the activity of virus early promoter[J]. China Oncology, 2017, 27(2): 109-114.
宋　丹,史　茜,侯向前,等. HPV-16 DOI： 10.19401/j.cnki.1007-3639.2017.02.005.

宋　丹, 史　茜, 侯向前. The effect of mutations in the upstream regulatory region of HPV-16 on the activity of virus early promoter[J]. China Oncology, 2017, 27(2): 109-114. DOI： 10.19401/j.cnki.1007-3639.2017.02.005.

摘要

背景与目的：新疆是宫颈癌高发区，该地区宫颈癌高发与人乳头瘤病毒16型(human papillomavirus type 16，HPV-16)感染密切相关。该研究旨在分析新疆地区妇女宫颈病样组织中HPV-16上游调控区(upstream regulatory region，URR)的突变及其功能。方法：以新疆妇女子宫颈上皮非典型增生(cervical intraepithelial neoplasia，CIN)和宫颈癌病样组织标本DNA为模板，PCR扩增HPV-16 URR片段，PCR产物经测序比对，筛选代表性的URR突变体构建至pGL3-Basic载体，将其转染Vero细胞，48 h后检测荧光素酶活性，分析URR突变体启动子活性。结果：采用聚合酶链反应(polymerase chain reaction，PCR)获得了55个HPV-16 URR DNA片段，测序及序列分析发现44个突变位点，其中nt7192(G→T)、nt7433(-→T)、nt7435(C→G)和nt7863(A→-)4个位点的突变为所有序列共有，nt7520(G→A)位点的突变存在于54个样品中，剩余39个位点的突变存在于不同样品中。根据突变的位置、频率和程度，筛选出9个URR突变体分别克隆至pGL3-Basic中荧光素酶基因前并转染Vero细胞。荧光素酶活性分析表明，不同URR突变体的启动子活性差异较大，来源于宫颈癌的URR突变体启动子活性显著高于来源于CIN的URR突变体(P0.01)，部分宫颈癌URR突变体的启动子活性显著高于SiHa和Caski细胞来源的URR参照序列的启动子活性。结论：新疆地区分离的HPV-16URR发生多位点突变，其中部分突变增强了URR内部启动子的活性，导致HPV-16致癌活性增强。

Abstract

Background and purpose: The incidence of cervical cancer is rather high in Xinjiang

which is closely associated with the infection of human papilloma virus type 16 (HPV-16). The purpose of this study was to analyze the variants and function of HPV-16 upstream regulatory region (URR) in the tissues of cervical cancer biopsies from Xinjiang. Methods: The DNAs were extracted from the tissues of cervical epithelial atypical hyperplasia (CIN) and cervical cancer biopsies. HPV-16 URR segments were amplified by PCR. Based on the sequence analysis of the URR

the representative URR variants were selected and cloned into pGL3-Basic. The recombinant plasmids were transfected into Vero cell lines respetively. Luciferase activity of transfected cells was detected 48 h after transfection. Results: Fifty-five HPV-16 URR DNA fragments were obtained through PCR

and 44 mutations were found from the URR fragments. 4 of these mutations

including nt7192(G→T)

nt7433(- →T)

nt7435 (C→G) and nt7863 (A→-) occurred in all sequences. The mutation at nt7520 (G→A) occurred in 54 URR sequences

and the 39 other mutations were present in different samples. Based on the location and frequency of the mutations in the URR fragments

9 URR variants were selected and cloned into pGL3-Basic. Then the luciferase activity of the cells transfected with pGL3- URR plasmids was detected respectively. Promoter activity of URR mutants from cervical cancer are significantly higher than that of URR mutants from CIN (P0.01). Promoter activity of URR fragments from some cervical cancer was significantly higher than that of the URR fragments from SiHa and Caski cells. Conclusion: Multiple mutations occurred in HPV-16 URR of cervical cancer patients from Xinjiang. The promoter activity and carcinogenicity of some URR mutants have been enhanced.

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