HPV E6/E7 mRNA检测在HPV分型及宫颈疾病筛查中的作用研究
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摘要
目的探讨人乳头瘤病毒(HPV)E6/E7 m RNA检测在女性宫颈疾病筛查中的作用。方法选取2016年9月至2017年9月贵州医科大学附属医院妇产科门诊和住院部就诊的HPV阳性感染者287例,其中宫颈疾病患者267例[包括宫颈炎、宫颈上皮内瘤变(CIN)Ⅰ~Ⅲ、宫颈癌],阳性感染者中体检无疾病表现及病理学检查无宫颈病变者20例。取患者宫颈脱落细胞用支链DNA技术(简称b-DNA技术)检测高危HPV E6/E7 m RNA,患者同时行宫颈液基薄层细胞检测和HPV分型检测,对不同年龄、HPV分型、阴道细胞学分型(简称TBS分型)及疾病分级的HPV E6/E7 m RNA的诊断价值进行效能分析。结果在不同年龄段内E6/E7阳性和阴性所占比例差异有统计学意义,P<0.05,在年龄组≤30岁组和>50~60岁组,E6/E7阳性者所占比例最大,分别为85.40%和84.40%;而HPV E6/E7m RNA拷贝数在不同年龄组内差异没有统计学意义。E6/E7结果在单一感染和混合感染,以及单一感染和多重感染间差异无统计学意义;在不同感染型别中,只有HPV 16型随病变严重程度的增加其感染率呈递增趋势(P<0.05),其余HPV型与病变严重程度无明显相关性(P>0.05)。不同疾病分组间,E6/E7阳性率的分布差异有统计学意义,表现为无宫颈病变组(0)<宫颈炎(15.60%) Objective To explore the detection of human papillomavirus E6/E7 m RNA in the screening of women's cervical disease.Methods A total of 287 HPV-positive patients were selected from September 2016 to September 2017 in the Department of Obstetrics and Gynecology of the Affiliated Hospital of Guizhou Medical University,among whom 267 patients had cervical disease(including cervicitis,CINⅠ~CINⅢ and cervical cancer).Another 287 HPV-postitive patients were chosen,among whom 20 had no disease manifestations on physical examination or cervical lesions on pathological examination.The branched-chain DNA technique(b-DNA)was used to detect the high-risk HPV E6/E7 m RNA in cervical exfoliated cells.Liquid-based thin-layer cervical cytology and HPV genotyping assays were used to analyze the diagnostic value of HPV E6/E7 m RNA in different ages,HPV typing,vaginal cytology type(TBS type),and disease grading.Results The difference in the proportion of positive and negative E6/E7 in different age groups was statistically significant(P<0.05);the proportion of E6/E7-positive patients in the age group≤30 years and >50-60 years was the largest,being 85.40% and 84.40%,respectively.However,HPV E6/E7 m RNA copy number did not differ significantly among different age groups.The E6/E7 results also showed that there was no significant difference between single infection and mixed infection,or between single infection and multiple infections.Only HPV 16 showed an increasing rate of infection with the increasing severity of lesions(P<0.05),and there was no significant correlation between other HPV types and lesion severity in different infection types(P>0.05).The distribution of E6/E7-positive rates had statistical difference among different disease groups:no cercical lesions(0)
引文
[1]Cambell LM,Pitta DR,De Assis AM,et al.Retrieval of HPV oncogenes E6 and E7 m RNA from cervical specimens using a manual open technology protocol[J].Springer Plus,2013,473(2):1-7.
[2]董云灿,耿建祥,张劲松,等.1722例已婚女性宫颈细胞中人乳头状瘤病毒基因的分型[J].国际检验医学杂志,2012,33(7):817-818,820.
[3]Munagala R,Kausar H,Munjal C,et al.With aferin A induces p53-dependent apoptosis by repression of HPV oncogenes and up-regulation of tumor suppressor proteins in human cervical cancer cells[J].Carcinogenesis,2011,32(11):1697-170.
[4]Schiffman M,Wentzensen N.Human papillomavirus infection and the multistage carcinogenesis of cervical cancer[J].Cancer Epidemiol Biomarkers Prev,2013,22:553-560.
[5]贾政军,周玉春,胡蓉,等.HPV m RNA实时荧光定量PCR检测及其与宫颈癌发生的关系[J].实用预防医学,2012,19(3):354-358.
[6]Rampias T,Boutati E,Pectasides E,et al.Activation of Wnt signaling pathway by human papillomavirus E6 and E7 oncogenes in HPV16-positive oropharyngeal squamous carcinoma cells[J].Mol Cancer Res,2010,8(3):433-443.
[7]Shen Y,Gong J,He Y,et al.Quantivirus?HPV E6/E7 RNA 3.0assay(b DNA)is as sensitive,but less specific than hybrid capture 2 test[J].J Virol Methods,2013,187(2):288-293.
[8]Haensen G,Krause U,Becker A,et al.Tumor hypoxia,p53,and prognosis in cervical cancer[J].Int J Radiation Oncol Biolphys,2001,50(4):865-872.
[9]Kraus I,Molden T,Holm R,et al.Presenee of E6 and E7 m RNA from human papillomavirus types 16,18,31,33 and 45 in the majority of cervical carcinomas[J].Clin Microbiol,2006,44:1310-1331.
[10]许剑利,徐克惠.高危型HPV检测及TCT检查在宫颈癌筛查中的应用分析[J].中国实用妇科与产科杂志,2014,30(12):946-949.
[11]吴云燕,裘明利,女性高危型人乳头瘤病毒感染分布情况研究[J].中国实用妇科与产科杂志,2017,33(1):122-124.
[12]Bosch FX,Manos MM,Muňoz N,et al.Prevalence of human papillomavirus in cervical cancer:a worldwide perspective[J].JNCI,1995,87(11):796-802.
[13]Ghittoni R,Accardi R,Hasan U,et al.The biological properties of E6 and E7 oncoproteins from human papillomaviruses[J].Virus Genes,2010,40(1):1-13.
[14]Steben M.Human papillomavirus infection:epidemiology and pathophysiology[J].Gynecol Oncol,2007,107(2 Suppl 1):S2-5.
[15]关嵩青,林莉,叶菲,等.深圳市妇女宫颈癌组织中人乳头瘤病毒16型E6、E7致癌基因的序列分析[J].中国实验诊断学,2014,18(3):397-400.
[16]耿建祥,王旭波.人乳头瘤病毒检测及其临床应用[M].北京:人民卫生出版社,2009:428-448.
[17]隋龙,丛青.妇科肿瘤筛查[J].中国实用妇科与产科杂志,2016,32(5):395-398.
[18]吴婷婷,李智,李丹军,等.高级别宫颈上皮内瘤变宫颈锥切术后病灶残留的多因素分析及临床处理探讨[J].中国实用妇科与产科杂志,2016,7(32):671-675.
[19]Ho GY,Studentsov YY,Bierman R,et al.Natural history of human papillomavirus type 16 virus-like partical antibodies in young women[J].Cancer Epidemiol Biomarkers Prev,2004,13:110-116.
[2]董云灿,耿建祥,张劲松,等.1722例已婚女性宫颈细胞中人乳头状瘤病毒基因的分型[J].国际检验医学杂志,2012,33(7):817-818,820.
[3]Munagala R,Kausar H,Munjal C,et al.With aferin A induces p53-dependent apoptosis by repression of HPV oncogenes and up-regulation of tumor suppressor proteins in human cervical cancer cells[J].Carcinogenesis,2011,32(11):1697-170.
[4]Schiffman M,Wentzensen N.Human papillomavirus infection and the multistage carcinogenesis of cervical cancer[J].Cancer Epidemiol Biomarkers Prev,2013,22:553-560.
[5]贾政军,周玉春,胡蓉,等.HPV m RNA实时荧光定量PCR检测及其与宫颈癌发生的关系[J].实用预防医学,2012,19(3):354-358.
[6]Rampias T,Boutati E,Pectasides E,et al.Activation of Wnt signaling pathway by human papillomavirus E6 and E7 oncogenes in HPV16-positive oropharyngeal squamous carcinoma cells[J].Mol Cancer Res,2010,8(3):433-443.
[7]Shen Y,Gong J,He Y,et al.Quantivirus?HPV E6/E7 RNA 3.0assay(b DNA)is as sensitive,but less specific than hybrid capture 2 test[J].J Virol Methods,2013,187(2):288-293.
[8]Haensen G,Krause U,Becker A,et al.Tumor hypoxia,p53,and prognosis in cervical cancer[J].Int J Radiation Oncol Biolphys,2001,50(4):865-872.
[9]Kraus I,Molden T,Holm R,et al.Presenee of E6 and E7 m RNA from human papillomavirus types 16,18,31,33 and 45 in the majority of cervical carcinomas[J].Clin Microbiol,2006,44:1310-1331.
[10]许剑利,徐克惠.高危型HPV检测及TCT检查在宫颈癌筛查中的应用分析[J].中国实用妇科与产科杂志,2014,30(12):946-949.
[11]吴云燕,裘明利,女性高危型人乳头瘤病毒感染分布情况研究[J].中国实用妇科与产科杂志,2017,33(1):122-124.
[12]Bosch FX,Manos MM,Muňoz N,et al.Prevalence of human papillomavirus in cervical cancer:a worldwide perspective[J].JNCI,1995,87(11):796-802.
[13]Ghittoni R,Accardi R,Hasan U,et al.The biological properties of E6 and E7 oncoproteins from human papillomaviruses[J].Virus Genes,2010,40(1):1-13.
[14]Steben M.Human papillomavirus infection:epidemiology and pathophysiology[J].Gynecol Oncol,2007,107(2 Suppl 1):S2-5.
[15]关嵩青,林莉,叶菲,等.深圳市妇女宫颈癌组织中人乳头瘤病毒16型E6、E7致癌基因的序列分析[J].中国实验诊断学,2014,18(3):397-400.
[16]耿建祥,王旭波.人乳头瘤病毒检测及其临床应用[M].北京:人民卫生出版社,2009:428-448.
[17]隋龙,丛青.妇科肿瘤筛查[J].中国实用妇科与产科杂志,2016,32(5):395-398.
[18]吴婷婷,李智,李丹军,等.高级别宫颈上皮内瘤变宫颈锥切术后病灶残留的多因素分析及临床处理探讨[J].中国实用妇科与产科杂志,2016,7(32):671-675.
[19]Ho GY,Studentsov YY,Bierman R,et al.Natural history of human papillomavirus type 16 virus-like partical antibodies in young women[J].Cancer Epidemiol Biomarkers Prev,2004,13:110-116.