外阴HPV感染表型与p53、p73多态性的相关性研究

发布时间：2018-04-09 10:09

本文选题：人乳头瘤病毒　切入点：鲍恩样丘疹病　出处：《北京协和医学院》2012年博士论文

【摘要】：尖锐湿疣(condyloma accuminatum CA)和鲍恩样丘疹病bowenoid papulosis of vulva, BP),前者是皮肤科门诊和性病门诊最常见的性病,由高危HPV感染者有癌变可能；后者病理形态上有组织排列和细胞的异型性,被认为是一种低度恶性的病毒感染性皮肤病或癌前病变。 HPV感染可以引起不同的临床表型,CA感染以低危型HPV为主,而BP感染的HPV以高危型为多。高危和低危型HPV感染的易感性在人群中是否有差异,感染高危低危型HPV之后其临床表型是否和人群的基因多态性有关,值得研究。近年关于p53及其同家族的抑癌基因p73的多态性和HPV相关的肿瘤易感性研究逐渐增多。部分研究发现HPV相关肿瘤和p53codon72密码子或p73第二外显子第4和14位碱基的基因多态性有关。但不同地区和人群的研究结果不一致。我们收集江苏南京地区感染HPV的尖锐湿疣和鲍恩样丘疹病患者的皮损和血样,以及在性病门诊进行健康体检的性病高危人群为阴性对照,进行HPV分型以及HPV感染临床表型和宿主p53、p73基因多态性的关系研究。初步探索不同HPV感染的临床表现是否有不同,以及HPV易感性和宿主p53、p73基因多态性的相关性,为今后的研究做一定的参考。全文分三部分。第一部分尖锐湿疣及鲍恩样丘疹病中HPV病毒基因型与疾病临床表现之间关系的研究目的通过直接测序法对尖锐湿疣以及鲍恩样丘疹病中HPV进行基因分型研究。根据测序所得到的型别,结合文献中尖锐湿疣和鲍恩样丘疹病中HPV的常见型别,分别设计型特异引物,通过特异性PCR来验证和补充测序结果,以探讨尖锐湿疣和鲍恩样丘疹病中HPV的分子流行病学状况。方法通过直接测序法对收集到的尖锐湿疣和鲍恩样丘疹病患者皮损中的HPV进行基因分型研究,根据测序得到的型别和文献中的常见型别,设计特异性引物,通过型特异性PCR来验证和补充测序结果,结合病例的临床表现,分析HPV不同型别感染的临床表现是否有不同。结果共收集CA标本124例,BP标本14例.CA患者HR-HPV的发生率为6.03%(7/116),BP为50%(7/14),两组比较,差异有统计学意义(X2=20.76P0.01)。这与既往文献报道的BP以HPV-16感染为主一致。因此,临床上怀疑BP的患者,除做病理确诊外,建议行HPV型别检测。感染HR-HPV的7例CA患者中,男性5例,女性2例；肛周3例,系带旁1例,包皮1例,小阴唇2例；菜花状5例,丘疹状1例,乳头样1例。经统计,性别、部位、皮损形态,与易感HR-HPV之间并无相关性, 结论我们对收集到的生殖器部位尖锐湿疣患者的HPV型别进行了鉴定,并对其皮损形态、皮损发生部位和感染HPV型别之间进行了统计学分析,发现前两者和患者感染HPV型别之间统计学上并无显著相关性。第二部分尖锐湿疣和鲍恩样丘疹病患者p53基因多态性分布的研究目的收集尖锐湿疣和鲍恩样丘疹病患者的血液标本,做p53多态性检测,分析p53的72密码子的基因改变与皮肤HPV感染的HPV型别、临床表现的关系。方法实验组选取本院皮肤科门诊2010年6月到2011年9月接受激光治疗的尖锐湿疣患者及病理确诊并接受激光治疗的鲍恩样丘疹病患者的血样及皮损。阴性对照组来自本院性病科性病体检排除尖锐湿疣的健康人群或患其他性病患者血样。实验组和对照组血样做p53多态性检测。结合患者皮损内感染HPV的型别,和实验组对照组p53多态性结果,对数据结果进行统计学分析。结果收集CA患者血样116例,测序阳性112例,收集BP患者血样14例,测序阳性14例。结果发现CA, BP患者体细胞有三种类型,CGC纯合型,CCC纯合型,CGC/CCC杂合型。CA组与对照组相比,Arg/Pro基因型者罹患CA的几率较其他两种纯和型低。BP组三种基因型与BP患病率无显著相关性(P0.05)。p53基因多态性与HPV高危和低危型易感性之间无显著相关。结论我们发现Arg/Pro基因型对罹患CA有保护性作用,但对同样为HPV感染高度相关的BP则未发现显著相关,将p53codon72Arg和Pro两个等位基因的分布与感染HPV的类型作相关性分析,未发现显著差异。究其原因,可能与我们所收集的BP病例数量较少有关,后续研究还需要进一步扩大样本量,完善临床数据,进一步进行研究。第三部分尖锐湿疣和鲍恩样丘疹病患者p73基因多态性分布及与HPV感染型别相关性的研究目的将尖锐湿疣和鲍恩样丘疹病患者的血液标本做p73多态性检测,分析p73第二外显子第4位和14位碱基的多态性与皮肤HPV感染的HPV型别、临床表现的关系。方法实验组选取本院皮肤科门诊2010年6月到2011年9月接受激光治疗的尖锐湿疣患者及病理确诊并接受激光治疗的鲍恩样丘疹病患者的血样及皮损。阴性对照组来自本院性病科性病体检排除尖锐湿疣的健康人群或患其他性病患者血样。实验组和对照组血样做p73多态性检测。结合患者皮损内感染HPV的型别,和实验组对照组p73多态性结果,对数据结果进行统计学分析。结果收集CA患者血样90例测序阳性83例,BP患者血样14例,测序阳性14例。结果发现BP、CA患者体细胞有A4T14/G4C14杂合型,A4T14纯合型,G4C14纯合型三种类型。p73第二外显子GC、AT两个等位基因的分布、与CA/BP相关性,以及与感染HPV的类型作相关性分析。在CA患者中GC和AT两基因分布与对照组相比无明显差异,BP患者中相比携带AT基因,携带GC基因则降低了BP的患病风险。与AT/AT基因相比,携带GC/AT和GC/GC基因降低了感染HPV患尖锐湿疣的风险。鲍恩样丘疹病组与对照组相比,携带A4T14基因型的个体罹患BP的风险要高于携带G4C14者；相对A4T14/A4T14基因型,G4C14/G4C14基因型罹患BP的风险显著降低；将G4C14/A4T14与G4C14/G4C14基因型相比,后者也有较低的罹患BP的风险。p73第二外显子的等位基因和基因型分布与感染高危和低危型HPV之间未见显著相关性。结论携带AT基因增加了BP和CA的患病风险。A4T14/A4T14基因型和G4C14/A4T14基因型比G4C4/G4C4基因型罹患BP的风险增加；携带GC/AT和GC/GC基因比携带AT/AT基因者患尖锐湿疣的风险较低。p73第二外显子的等位基因和基因型分布与CA和BP中高危和低危型HPV易感性之间在本次研究中未见显著相关性。
[Abstract]:Condyloma Acuminatum CA and Bowen ' s papulosis , the former is the most common sexually transmitted disease in Dermatology Clinic and STD clinic , which may be caused by high - risk HPV infection .
The latter pathological form is characterized by tissue arrangement and cell differentiation , and is considered to be a low - degree malignant virus infectious skin disease or precancerous lesion .

HPV infection can lead to different clinical phenotypes , CA infection is mainly low - risk HPV , while the HPV of BP infection is high risk type . The susceptibility of HPV infection after high - risk and low - risk HPV infection is related to the polymorphism of human gene polymorphism . In recent years , it has been found that HPV - related tumor and p53codon72 codon or p73 - second exon 4 and 14 - base gene polymorphism are related . However , the results of different regions and population are inconsistent .

We collected lesions and blood samples from patients with HPV infection in Nanjing , Jiangsu Province , as well as the relationship between HPV typing and clinical phenotype of HPV infection and p53 and p73 gene polymorphism in STD clinic .

Study on the relationship between HPV genotypes and clinical manifestations in first partial condyloma and Bowen - like papulosis

Objective To study the genotyping of HPV in patients with condyloma and Bowen ' s papulosis by direct sequencing . According to the type of HPV in the literature , specific primers were designed to verify and complement the sequencing results by specific PCR to investigate the molecular epidemiology of HPV in Condyloma Acuminatum and Bowen - like papulosis .

Methods HPV types in lesions of patients with condyloma and Bowen - like papulosis were analyzed by direct sequencing . Specific primers were designed according to the types of sequencing and the common types in the literature . The results of sequencing were verified and supplemented by typing specific PCR . The clinical manifestations of the cases were analyzed .

Results The incidence of HR - HPV was 6.03 % ( 7 / 116 ) , BP was 50 % ( 7 / 14 ) .
3 cases were perianal , 1 case was near the frenulum , 1 case was covered , 2 cases were small .
There were 5 cases of cauliflower , 1 case of papule , 1 case of nipple , and no correlation was found between statistic , sex , site , lesion morphology and susceptibility HR - HPV .

Conclusion The HPV types of patients with genital area of genital area were identified , and the morphology , location of lesions and HPV types were analyzed statistically . There was no statistically significant correlation between the HPV types before and after the detection of HPV types .

Study on p53 gene polymorphism in the second part of patients with Condyloma Acuminatum and Bowen - like papulosis

Objective To investigate the relationship between the changes of p53 gene and HPV type and clinical manifestation of HPV infection in patients with condyloma and Bowen - like papulosis .

Methods The blood samples and skin lesions of the patients who were diagnosed and treated with laser were selected from the dermatological department of our hospital from June 2010 to September 2011 .

Results There were 116 cases of blood samples collected from CA patients , 112 cases were sequenced , 14 cases were collected from BP and 14 cases were sequenced .

Conclusion We find that the Arg / Pro genotype has protective effect on CA , but there is no significant correlation between the distribution of p53codon72Arg and Pro two alleles and the type of infected HPV . There is no significant difference between the distribution of p53codon72Arg and Pro .

The distribution of p73 gene polymorphism in patients with third partial condyloma and Bowen - like papulosis and its correlation with HPV infection

Objective To investigate the polymorphism of p73 gene exon 4 and 14 base and the relationship between HPV type and clinical manifestation of HPV infection .

Methods The blood samples and skin lesions of the patients who received laser therapy from June 2010 to September 2011 were selected from the experimental group . The negative control group received blood samples from the healthy population or patients with other sexually transmitted diseases . The control group and the control group were tested for p73 polymorphism .

Results There were 83 cases of blood sample in CA patients , 14 cases of BP and 14 cases of sequencing . The results showed that there was no significant difference between the two alleles of the two alleles in the patients with CA , the correlation between the two alleles of A4T14 / G4C14 , the correlation with CA / BP and the type of infected HPV .
Compared with A4T14 / A4T14 genotype , the risk of G4C14 / G4C14 genotype was significantly lower than that of A4T14 / A4T14 .
Comparing G4C14 / A4T14 with G4C14 / G4C14 genotype , the latter also had lower risk of BP . The allele and genotype distribution of p73 second exon was not significantly correlated with high - risk and low - risk HPV infection .

Conclusion The genotypes of A4T14 / A4T14 and G4C14 / A4T14 were higher than those of G4C4 / G4C4 genotype .
There was no significant correlation between allele and genotype distribution of p73 second exon and high - risk and low - risk HPV susceptibility in CA and BP .

【学位授予单位】：北京协和医学院
【学位级别】：博士
【学位授予年份】：2012
【分类号】：R752.53

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