云南地区宫颈癌癌前病变患者HPV 56的分子流行病学调查

周紫云; 王莹莹; 严志凌; 赵凌锋; 杨希; 许金美; 张芍

doi:10.12259/j.issn.2095-610X.S20231005

云南地区宫颈癌癌前病变患者HPV 56的分子流行病学调查

doi: 10.12259/j.issn.2095-610X.S20231005

1.
曲靖市疾病预防控制中心，云南曲靖　655000
2.
昆明医科大学基础医学院，云南昆明　650500
3.
昆明医科大学第三附属医院妇科，云南昆明　650118

基金项目: 云南省科技厅-昆明医科大学应用基础研究联合专项基金资助项目（202201AY070001-139）

详细信息

作者简介:
周紫云（1993～），女，云南师宗人，理学硕士，主要从事病毒分子流行病学研究工作

王莹莹与周紫云对本文有同等贡献

张芍，昆明医科大学第三附属医院妇科，硕士研究生，主治医师。中国抗癌协会宫颈癌专委会成员；昆明医科大学“一对一”本科生导师。主要从事普通妇科、妇科肿瘤、女性生殖内分泌疾病等研究工作；尤其擅长妇科常见良恶性肿瘤的手术、放化疗、内分泌及靶向治疗等综合治疗。近5年主持云南省科技厅面上项目在研课题：VRK1通过磷酸化P53蛋白调控细胞周期及凋亡通路促进宫颈癌发生发展的分子机制研究（202201AY070001-140）。参加国家自然科学基金、云南省科技厅、教育厅多项课题。在中文北大核心期刊以第一作者发表论文4篇；参与发表SCI 论文4篇。荣获2022年云南省卫生科技成果奖励三等奖（宿主免疫相关基因及HPV变异与宫颈癌发生发展的相关性和诊断价值）

通讯作者:
张芍，E-mail：zsspoon@126.com

中图分类号: R737.3
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出版历程
- 收稿日期: 2023-03-12
- 网络出版日期: 2023-09-11
- 刊出日期: 2023-10-25

Molecular Epidemiological Investigation of HPV 56 in Precancerous Cervical Cancer Patients in Yunnan Province

1.
Center for Disease Control and Prevention，Qujing City，Qujing Yunnan 655000
2.
School of Basic Medicine，Kunming Medical University，Kunming Yunnan 650500
3.
Dept. of Gynaecologic Oncology，The 3rd Affiliated Hospital of Kunming Medical University， Kunming Yunnan 650118，China

摘要

摘要: 目的研究单一感染HPV56型的癌前病变患者中HPV56 E6、E7和L1基因变异特征，获取云南地区宫颈癌癌前病变患者中HPV56型分子流行病学数据。方法随机选取300例宫颈癌癌前病变患者，通过HPV分型试剂盒对患者感染HPV的型别进行检测，并对HPV56型感染患者中HPV E6 E7和L1基因进行测序，最后完成对研究对象所感染的HPV56的谱系分布及HPV56 E6、E7和L1基因的遗传变异进行分析。结果研究对象所感染的HPV56病毒主要为A谱系，90%以上为A谱系，约10%为B谱系，HPV56 E6、E7、L1基因中共存在13个变异位点，其中E6基因中6个、E7基因中4个、L1基因中3个。HPV56 E6基因的6个变异位点中有2个为同义突变（T158C（S19S）、G437A（P112P）），4个为非同义突变（A141C（S14R）、A241G（N47S）、A263C（K54N）、G279A（D60N））。HPV56 E7基因的4个变异位点中均为非同义突变（C601A（D10E）、G605A（V12I）、G770C（E67Q）、G802C（Q77H））。HPV56 L1基因的3个变异位点中有2个为同义突变（G6454T（P321P）、A6868C（I459I）），1个为非同义突变（A5867G（N126D））。结论初步获得了云南地区宫颈癌癌前病变患者中HPV56 E6、E7和L1基因变异情况，获得了HPV56 E6、E7和L1基因的分子流行学特征，为进一步分析云南地区宫颈癌癌前病变患者中HPV56 E6、E7和L1基因的分子流行病学特征提供了必要的数据。
- 宫颈癌癌前病变 /
- HPV56 /
- 变异 /
- 谱系
Abstract: Objective To study the genetic variation characteristics of HPV56 E6, E7 and L1 in patients with precancerous lesions infected with HPV56, and to obtain the molecular epidemiological data of HPV56 in patients with precancerous lesions of cervical cancer in Yunnan. Methods A total of 300 cervical intraepithelial neoplasia patients were enrolled and HPV genotyping kit was used to detect the types of HPV infection in the patients. The HPV E6, E7, and L1 genes in patients infected with HPV 56 were sequenced. Finally, the lineage distribution of HPV 56 and the genetic variations of HPV 56 E6, E7, and L1 genes in the study subjects were analyzed. Results In this study, the HPV56 virus infected by the study subjects mainly belonged to the A lineage, with more than 90% belonged to the A lineage, and about 10% belonged to the B lineage. There were 13 mutation loci in the E6, E7 and L1 genes of HPV56, including 6 in the E6 gene, 4 in the E7 gene and 3 in the L1 gene. Among the 6 mutation sites of HPV56 E6 gene, 2 were synonymous mutations （T158C （S19S）, G437A （P112P））, and 4 were non-synonymous mutations （A141C （S14R）, A241G （N47S）, A263C （K54N）, G279A （D60N））. The four mutation sites of HPV56 E7 gene were all non-synonymous mutations （C601A （D10E）, G605A （V12I）, G770C （E67Q）, G802C （Q77H））. Among the three mutation sites of HPV56 L1 gene, two were synonymous mutations （G6454T （P321P）, A6868C （I459I））, and one was non-synonymous mutation （A5867G （N126D））. Conclusion This study obtained preliminary data on the genetic variations of HPV 56 E6, E7, and L1 genes in CIN patients in Yunnan area, and obtained the molecular epidemiological characteristics of HPV 56 E6, E7, and L1 genes. It provides necessary data for further analysis of the molecular epidemiological characteristics of HPV 56 E6, E7, and L1 genes in CIN patients in Yunnan area.
- Cervical intraepithelial neoplasia /
- HPV56 /
- Variation /
- Lineage

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图 1 HPV56 E6基因系统发生树

Figure 1. Phylogenetic tree of E6 of HPV56

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图 2 HPV56 E7基因系统发生树

Figure 2. Phylogenetic tree of E7 of HPV56

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图 3 HPV56 L1基因系统发生树

Figure 3. Phylogenetic tree of L1 of HPV56

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表 1 HPV56 E6、E7、L1基因的扩增引物和测序引物

Table 1. The amplification primers and sequencing primers of E6 E7 and L1 genes of HPV56

基因	方向	序列5′-3′	退火温度℃
E6	Forward	ATTGGGAGTGACCGAAAAGG	63
	Reverse	ACAACACGCAGGTCCTCTTT
E7	Forward	GCTACAGATGTCAAAGTCCG	61
	Reverse	GCCTCTACTTCAAACCATCC
L1-1	Forward	GCCCCTTTAGGTAATGTGTGG	63
	Reverse	CCACATAGAATCACCATAGGCAT
L1-1	Forward	ATGATAGACACAGGATTTGGCG	63
	Reverse	ATACAACACACAAACACAGTTACAC

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表 2 HPV56 E6、E7、L1基因变异情况

Table 2. E6，E7 and L1 gene variation of HPV56

核苷酸位置	E6						E7				L1
核苷酸位置	141	158	241	263	279	437	601	605	770	802	5867	6454	6868
X74483.1-E6	A	T	A	A	G	G	C	G	G	G	A	G	A
cin1-2	−	C	−	−	A	A	A	−	C	C	G	T	C
cin1-4	−	C	−	−	A	A	A	−	C	−	G	T	C
cin1-5	−	C	−	−	A	A	A	−	C	−	0	0	0
cin1-6	−	C	−	−	A	A	A	−	C	−	0	0	0
cin1-7	−	C	−	−	A	A	A	−	C	−	G	T	C
cin1-8	−	C	−	−	A	A	A	−	C	−	G	T	C
cin1-9	−	C	−	−	A	A	A	−	C	−	G	T	C
cin1-10	−	C	−	−	A	A	A	−	C	−	G	T	C
cin1-12	−	C	−	−	A	A	A	−	C	−	G	T	C
cin1-14	−	C	−	−	A	A	A	−	C	−	G	T	C
cin1-16	−	C	−	−	A	A	A	−	C	−	G	T	C
cin1-17	−	C	−	−	A	A	0	0	0	0	G	T	C
cin1-19	−	C	−	−	A	A	0	0	0	0	0	0	0
cin1-20	−	C	−	−	A	A	0	0	0	0	G	T	C
cin1-21	0	0	0	0	0	0	0	0	0	0	G	T	C
cin1-22	0	0	0	0	0	0	0	0	0	0	G	T	C
cin2-2	0	0	0	0	0	0	A	−	C	−	0	0	0
cin2-5	0	0	0	0	0	0	A	−	C	−	0	0	0
cin2-12	0	0	0	0	0	0	A	−	C	−	0	0	0
cin2-16	−	C	−	−	A	A	A	−	C	−	G	T	C
cin2-18	−	C	−	−	A	A	A	−	C	−	0	0	0
cin2-19	−	C	−	−	A	A	A	−	C	−	G	T	C
cin2-22	0	0	0	0	0	0	A	−	C	−	0	0	0
cin2-28	−	C	−	−	A	A	0	0	0	0	G	T	C
cin2-29	−	C	−	−	A	A	0	0	0	0	0	0	0
cin2-32	−	C	−	−	A	A	0	0	0	0	G	T	C
cin2-33	−	C	−	−	A	A	A	−	C	−	0	0	0
cin2-34	−	C	−	−	A	A	0	0	0	0	0	0	0
cin2-35	−	C	−	−	A	A	A	−	C	−	G	T	C
cin2-37	0	0	0	0	0	0	A	−	C	−	G	T	C
cin2-42	C	−	G	C	−	−	−	A	−	C	0	0	0
cin2-46	C	−	G	C	−	−	−	A	−	C	0	0	0
氨基酸变化	S14R	−	N47S	K54N	D60N	−	D10E	V12I	E67Q	Q77H	N126D	−	−
标记为“0”的部分表示未扩增/测出基因序列; 标记为“-”的部分表示未检测到变异。

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参考文献(28)

[1]	Arbyn M,Weiderpass E,Bruni L,et al. Estimates of incidence and mortality of cervical cancer in 2018: A worldwide analysis[J]. Lancet Glob Health,2020,8(2):e191-e203. doi: 10.1016/S2214-109X(19)30482-6
[2]	李双,李明珠,丛青,等. 人乳头瘤病毒疫苗临床应用中国专家共识[J]. 中国妇产科临床杂志,2021,22(2):225-234. doi: 10.13390/j.issn.1672-1861.2021.02.045
[3]	Burd E M. Human papillomavirus and cervical cancer[J]. Clin Microbiol Rev.,2003,16(1):1-17. doi: 10.1128/CMR.16.1.1-17.2003
[4]	Lorincz A T, Quinn A P, Goldsborough M D, et al. Human papillomavirus type 56: A new virus detected in cervical cancers [J]. J Gen Virol, 1989, 70 （ Pt 11）: 3099-3104.
[5]	Yang L,Yang H,Wu K,et al. Prevalence of HPV and variation of HPV 16/HPV 18 E6/E7 genes in cervical cancer in women in South West China[J]. J Med Virol,2014,86(11):1926-1936. doi: 10.1002/jmv.24043
[6]	Zhao S,Zhao X,Hu S,et al. Distribution of high-risk human papillomavirus genotype prevalence and attribution to cervical precancerous lesions in rural North China[J]. Chin J Cancer Res,2019,31(4):663-672. doi: 10.21147/j.issn.1000-9604.2019.04.10
[7]	Harari A,Chen Z,Burk R D. Human papillomavirus genomics: Past,present and future[J]. Curr Probl Dermatol,2014,45:1-18.
[8]	Zhou Z,Yang H,Yang L,et al. Human papillomavirus type 16 E6 and E7 gene variations associated with cervical cancer in a Han Chinese population[J]. Infect Genet Evol,2019,73:13-20. doi: 10.1016/j.meegid.2019.04.008
[9]	Wu Y, Chen Y, Li L, et al. Analysis of mutations in the E6/E7 oncogenes and L1 gene of human papillomavirus 16 cervical cancer isolates from China [J]. J Gen Virol, 2006, 87（Pt 5）: 1181-1188.
[10]	Lee C W,Bae J H,Lee S J,et al. Distribution of human papillomavirus type 16 E6 and E7 gene variants in the progression of cervical dysplasia in Korean women[J]. J Obstet Gynaecol Res,2011,37(10):1320-1326. doi: 10.1111/j.1447-0756.2010.01520.x
[11]	Sun Z,Ren G,Cui X,et al. Genetic diversity of HPV-16 E6,E7,and L1 genes in women with cervical lesions in Liaoning Province,China[J]. Int J Gynecol Cancer,2011,21(3):551-558. doi: 10.1097/IGC.0b013e3182112023
[12]	Buck C B,Day P M,Trus B L. The papillomavirus major capsid protein L1[J]. Virology,2013,445(1-2):169-174. doi: 10.1016/j.virol.2013.05.038
[13]	Jing Y,Wang T,Chen Z,et al. Phylogeny and polymorphism in the long control regions E6,E7,and L1 of HPV Type 56 in women from southwest China[J]. Mol Med Rep,2018,17(5):7131-7141.
[14]	Burk R D,Harari A,Chen Z. Human papillomavirus genome variants[J]. Virology,2013,445(1-2):232-243. doi: 10.1016/j.virol.2013.07.018
[15]	中华医学会. 临床诊疗指南. 妇产科学分册[M]. 北京: 人民卫生出版社, 2007: 69-73.
[16]	Munoz N,Bosch FX,de Sanjose S,et al. Epidemiologic classification of human papillomavirus types associated with cervical cancer[J]. N Engl J Med,2003,348(6):518-527. doi: 10.1056/NEJMoa021641
[17]	Franceschi S,Rajkumar R,Snijders P J,et al. Papillomavirus infection in rural women in southern India[J]. Br J Cancer,2005,92(3):601-606. doi: 10.1038/sj.bjc.6602348
[18]	Nielsen A,Kjaer S K,Munk C,et al. Type-specific HPV infection and multiple HPV types: Prevalence and risk factor profile in nearly 12,000 younger and older Danish women[J]. Sex Transm Dis,2008,35(3):276-282. doi: 10.1097/OLQ.0b013e31815ac5c7
[19]	Liu J,Ma S,Qin C,et al. Prevalence and genotype distribution of human papillomavirus in Zhengzhou,China,in 2016[J]. Arch Virol,2020,165(3):731-736. doi: 10.1007/s00705-019-04515-3
[20]	Wu H,Wu E,Ma L,et al. Lineage distribution and E2 sequence variation of high-risk human papillomavirus types isolated from patients with cervical cancer in Sichuan Province,China[J]. Arch Virol,2015,160(11):2845-2855. doi: 10.1007/s00705-015-2559-z
[21]	Dai S,Yao Y,Yan Z,et al. The association of human papillomavirus type 16 E2 variations with cervical cancer in a Han Chinese population[J]. Infect Genet Evol,2018,64:241-248. doi: 10.1016/j.meegid.2018.07.002
[22]	Morgan E L,Scarth J A,Patterson M R,et al. E6-mediated activation of JNK drives EGFR signalling to promote proliferation and viral oncoprotein expression in cervical cancer[J]. Cell Death Differ,2020,28(5):1669-1687.
[23]	Scarth J A,Patterson M R,Morgan E L,et al. The human papillomavirus oncoproteins: A review of the host pathways targeted on the road to transformation[J]. J Gen Virol,2021,102(3):001540.
[24]	Howie H L,Katzenellenbogen R A,Galloway D A. Papillomavirus E6 proteins[J]. Virology,2009,384(2):324-334. doi: 10.1016/j.virol.2008.11.017
[25]	Hiller T,Stubenrauch F,Iftner T. Isolation and functional analysis of five HPVE6 variants with respect to p53 degradation[J]. J Med Virol,2008,80(3):478-483. doi: 10.1002/jmv.21093
[26]	Roman A,Munger K. The papillomavirus E7 proteins[J]. Virology,2013,445(1-2):138-168. doi: 10.1016/j.virol.2013.04.013
[27]	Kirnbauer R,Taub J,Greenstone H,et al. Efficient self-assembly of human papillomavirus type 16 L1 and L1-L2 into virus-like particles[J]. J Virol,1993,67(12):6929-6936. doi: 10.1128/jvi.67.12.6929-6936.1993
[28]	Tan W,Felber B K,Zolotukhin A S,et al. Efficient expression of the human papillomavirus type 16 L1 protein in epithelial cells by using Rev and the Rev-responsive element of human immunodeficiency virus or the cis-acting transactivation element of simian retrovirus type 1[J]. J Virol,1995,69(9):5607-5620. doi: 10.1128/jvi.69.9.5607-5620.1995

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云南地区宫颈癌癌前病变患者HPV 56的分子流行病学调查

doi: 10.12259/j.issn.2095-610X.S20231005

通讯作者:
张芍，E-mail：zsspoon@126.com

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Molecular Epidemiological Investigation of HPV 56 in Precancerous Cervical Cancer Patients in Yunnan Province

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目录

[1]	孟丽燕, 王超群, 陈星慧, 何军晶, 牛雅茹, 马丽莎. 早期宫颈癌术后HPV持续感染的危险因素分析, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20240422
[2]	张帆, 郭伟昌, 谭洪, 殷和佳, 彭露萍, 赵燕, 李会芳. 葡萄糖变异参数构建糖尿病周围神经病变风险预测模型的探讨, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20230526
[3]	谭丁及, 尹洪莉, 朱锐, 张曦, 余鑫, 飞勇, 李昕, 段铭, 何亮, 杨宏英. 宫颈上皮内瘤变和宫颈癌患者的阴道微生态特点, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20210821
[4]	张娟, 张红芸. 宫颈管搔刮术在宫颈癌筛查中的价值, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20210822
[5]	高健博, 史晓依, 郝小生, 梁建民, 吴雪梅, 陈银波. 儿童视神经脊髓炎谱系疾病16例临床分析, 昆明医科大学学报.
[6]	卿清, 刘洋, 周红林. 宫颈癌筛查策略的研究进展, 昆明医科大学学报.
[7]	廖婧, 许长俊, 周红林, 刘洋. c-Myc、hTERC基因检测及HC2-HPV分子检测对宫颈癌前病变的诊断价值, 昆明医科大学学报.
[8]	徐永琴, 梅雪, 罗汝琼, 李伟, 张海, 张方芳. HPV DNA含量及血清、分泌物MMP在宫颈癌患者中的表达及对预后的反应价值, 昆明医科大学学报.
[9]	翟淑娟, 潘景, 孙春意, 刘洋, 杨莹莹, 周红林. p62、OPTN在宫颈癌中的表达及其与HPV的相关性, 昆明医科大学学报.
[10]	孙春意, 翟淑娟, 曹青萍, 杨莹莹, 周红林. 宫颈柱状上皮异位与宫颈癌及CIN的关系, 昆明医科大学学报.
[11]	杨琳琳, 王薇, 张磊, 张红平, 杨宏英. 宫颈癌患者治疗前检测血清HPV16E6 mRNA和CK19 mRNA的临床意义, 昆明医科大学学报.
[12]	朱天波. 宫颈癌术前新辅助化疗的临床应用, 昆明医科大学学报.
[13]	李正金. 13种高危型HPV感染联合TCT检查在大理地区宫颈癌筛查中的应用, 昆明医科大学学报.
[14]	段敬. 血清SCCA及MMPs水平在宫颈癌患者检测宫颈癌的临床分析, 昆明医科大学学报.
[15]	严志凌. 宿主p53基因多态性与HPV16 感染、宫颈癌发生发展的相关性, 昆明医科大学学报.
[16]	马原. 腹腔镜下宫颈癌根治术治疗宫颈癌的临床分析, 昆明医科大学学报.
[17]	刘梅. 2007年至2008年昆明地区腹泻患儿轮状病毒非结构蛋白基因特征的分析, 昆明医科大学学报.
[18]	. 康复新外敷治疗宫颈癌放疗后皮肤损伤的疗效观察, 昆明医科大学学报.
[19]	曹兰. 儿童、青少年强迫谱系障碍所致皮肤损害54例临床分析, 昆明医科大学学报.
[20]	黄雅. Brn-3a、PPAR-γ在宫颈癌及癌前病变中的表达, 昆明医科大学学报.

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云南地区宫颈癌癌前病变患者HPV 56的分子流行病学调查

doi: 10.12259/j.issn.2095-610X.S20231005

通讯作者: 张芍，E-mail：zsspoon@126.com

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Molecular Epidemiological Investigation of HPV 56 in Precancerous Cervical Cancer Patients in Yunnan Province

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通讯作者:
张芍，E-mail：zsspoon@126.com