人乳头瘤病毒与宫颈病变发生发展的研究
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摘要
目的
1.研究人乳头瘤病毒(HPV)L1蛋白在液基细胞学结果异常涂片中的表达与临床意义。
2.探讨肿瘤相关基因14-3-3σ、hWAPL、HIF-1α的mRNA含量在正常宫颈组织、宫颈上皮内瘤变组织和宫颈癌组织中的表达量变化与意义。
3.探讨HC-Ⅱ法检测高危型HPV DNA病毒载量和宫颈病变的关系,HC-Ⅱ法在高度宫颈上皮内瘤变(HSIL)治疗后临床随诊的应用价值。
方法
1.对274例液基细胞学结果为ASC-US以上、HC-Ⅱ检测高危型HPV阳性的涂片进行HPV L1全反应抗体的免疫染色。所有病例均于初次细胞学检查后进行组织病理学检查。
2.收集正常宫颈、宫颈上皮内瘤变CIN1、CIN2、CIN3及未经放化疗的宫颈鳞癌组织新鲜标本。提取组织中的总RNA,应用实时荧光定量PCR法检测14-3-3σ、hWAPL、HIF-1σmRNA在正常宫颈、宫颈上皮内瘤变及宫颈鳞癌组织中的相对表达量。
3.2006年9月-2008年12月,对2830名妇女行宫颈癌筛查,以组织病理学检查结果为金标准,分析及HR-HPV病毒载量和宫颈病变的关系。
4.总结2004年1月至2008年6月经组织病理学证实≥CIN2,接受锥切术的648例患者相关资料。对术前、术后随访中HC-Ⅱ变化与病变残留复发情况进行动态观察和分析。
结果
1.不同细胞学涂片L1蛋白的表达情况不同,LSIL组L1表达率最高,达78.26%(90/115),其次为ASC-US,达63.81%(67/105),HSIL+SCC组为9.76%(4/41)。各组间统计有显著性差异(P<0.01)。L1表达者80%(44/55)消退至正常细胞学,20%(11/55)结果稳定,无一例进展。L1不表达者仅有37.50%(6/16)消退至正常细胞学,43.75%(7/16)结果稳定,细胞学结果进展占18.75%(3/16),进展为ASC-H和HSIL。
2.宫颈鳞癌组织中14-3-3σmRNA表达量高于正常宫颈组织及宫颈上皮内瘤变组织,统计学分析各组间有显著性差异(P<0.05)。
3.宫颈上皮内瘤变及宫颈鳞癌组织中hWAPLmRNA表达量均高于正常宫颈组织,统计学有显著性差异(P<0.05)。宫颈鳞癌组织中hWAPL mRNA表达量高于宫颈上皮内瘤变组织,统计学有显著性差异(P<0.05)。
4.正常宫颈、宫颈上皮内瘤变与宫颈鳞癌组织中HIF-1αmRNA的含量没有显著性差异(P>0.05)。
5.高危型HPV DNA病毒载量在不同宫颈病变中差异无显著性。
6.锥切术后切缘阳患者持续或复发≥CIN2为20.63%(20/97)明显高于切缘阴患者的1.63%(9/551),p<0.01。锥切术后HC-Ⅱ结果持续阴性,复发例数为0,HC-Ⅱ结果持续阳性,复发率达25%(23/92)。
结论
1.L1蛋白在液基细胞涂片中的表达情况能够帮助了解宫颈病变的进展程度。HPVL1不表达或持续不表达者,提示病变进展。
2.宫颈癌前病变进展为宫颈癌时,14-3-3σ可能参与其过程。
3.hWAPL可能参与了宫颈上皮内瘤变的发生、发展、癌变的过程。
4.HR-HPV DNA病毒载量与宫颈病变的级别没有相关性,RLU/PC≥1即有临床意义,定性比定量更重要。
5.HC-Ⅱ检测对锥切术后随访有密切关系。
OBJECTIVES
1.To investigate whether it is possible to use detection of the human papillomavirus (HPV) L1 capsid protein to predict the course of mild or moderate cervical intraepithelial neoplasia(CIN).
2.To explore the expressions of 14-3-3σ,hWAPL and HIF-1αmRNA in cervical cancer and CINs。To analyze the role they play in the progression of CIN_s.
3.To observe the relation between cervical neoplasia and the viral load aquired by HCII. To assess the value of HC-Ⅱfor follow up after conization.
METHODS
1.Imrnunocytochemical analysis using antibody against HPV L1 capsid protein was carried out on 274 cases of TRIPATH Pap tests with abnormal cytologic diagnoses and positive for high-risk HPV DNA detected by HC-Ⅱ.
2.Collect fresh tissues from cervix.Extract total RNA and evaluate the relative expressions of 14-3-3σ,hWAPL and HIF-1αmRNA with real-time fluorogenetic quantitative PCR(RFQ-PCR).
3.A total of 2830 women were screened for cervical lesion.We assessed the relations between CINs and the viral load aquired by HC-Ⅱ.
4.648 patients accepted conization for≥CIN II histologically.We analyzed the changes of HC-Ⅱresults and the recurrences of lesions.
RESULTS
1.Cytologic diagnosis revealed a higher expression of L1 protein in LSIL(75.63%, 90/119)than in ASC-US(63.81%,67/105)and in HSIL/SCC(4/41,9.76%)(P<0.01).
2.71 cases without treatment accepted LCT test after lyear.In the cases expressing L1, 80%(44/55) regressed to NILM,and no one progressed.In the cases not expressing L1,only 37.50%(6/16) regressed to NILM,and 18.75%(3/16) progressed to ASC-H and HSIL.
3.The average quantity of 14-3-3σmRNA in cervical caner tissues was significantly higher than that in cervicitis tissues(t-test,P<0.05).
4.The average quantity of hWAPL mRNA in cervical caner and CINs tissues was significantly higher than that in cervicitis tissues(t-test,P<0.05).
5.The viral loads of high-risk HPV DNA in invasive cervical cancer and CINs were similar.
6.After conization,20.62%of the patients with positive surgical margins got persistent/recurrent lesions,while only 1.63%of those with negative surgical margins did.No persistent/recurrent lesions happened on the patients with persistent negative results of HC-Ⅱ.While 25%of those with persistent positive results of HC-Ⅱdid.
CONCLUSIONS
1.The expression of HPV L1 in LSIL was significantly higher than in HSIL.The disease may progress while not expressing L1 protein or non-expressing persistently.
2.14-3-3σand hWAPL may play an role in the emergence of cervical cancer.
3.The viral load of high-risk HPV DNA has no relation with the severity of cervical lesions.HC-Ⅱhas great significance for the follow-up after conization.
1.研究人乳头瘤病毒(HPV)L1蛋白在液基细胞学结果异常涂片中的表达与临床意义。
2.探讨肿瘤相关基因14-3-3σ、hWAPL、HIF-1α的mRNA含量在正常宫颈组织、宫颈上皮内瘤变组织和宫颈癌组织中的表达量变化与意义。
3.探讨HC-Ⅱ法检测高危型HPV DNA病毒载量和宫颈病变的关系,HC-Ⅱ法在高度宫颈上皮内瘤变(HSIL)治疗后临床随诊的应用价值。
方法
1.对274例液基细胞学结果为ASC-US以上、HC-Ⅱ检测高危型HPV阳性的涂片进行HPV L1全反应抗体的免疫染色。所有病例均于初次细胞学检查后进行组织病理学检查。
2.收集正常宫颈、宫颈上皮内瘤变CIN1、CIN2、CIN3及未经放化疗的宫颈鳞癌组织新鲜标本。提取组织中的总RNA,应用实时荧光定量PCR法检测14-3-3σ、hWAPL、HIF-1σmRNA在正常宫颈、宫颈上皮内瘤变及宫颈鳞癌组织中的相对表达量。
3.2006年9月-2008年12月,对2830名妇女行宫颈癌筛查,以组织病理学检查结果为金标准,分析及HR-HPV病毒载量和宫颈病变的关系。
4.总结2004年1月至2008年6月经组织病理学证实≥CIN2,接受锥切术的648例患者相关资料。对术前、术后随访中HC-Ⅱ变化与病变残留复发情况进行动态观察和分析。
结果
1.不同细胞学涂片L1蛋白的表达情况不同,LSIL组L1表达率最高,达78.26%(90/115),其次为ASC-US,达63.81%(67/105),HSIL+SCC组为9.76%(4/41)。各组间统计有显著性差异(P<0.01)。L1表达者80%(44/55)消退至正常细胞学,20%(11/55)结果稳定,无一例进展。L1不表达者仅有37.50%(6/16)消退至正常细胞学,43.75%(7/16)结果稳定,细胞学结果进展占18.75%(3/16),进展为ASC-H和HSIL。
2.宫颈鳞癌组织中14-3-3σmRNA表达量高于正常宫颈组织及宫颈上皮内瘤变组织,统计学分析各组间有显著性差异(P<0.05)。
3.宫颈上皮内瘤变及宫颈鳞癌组织中hWAPLmRNA表达量均高于正常宫颈组织,统计学有显著性差异(P<0.05)。宫颈鳞癌组织中hWAPL mRNA表达量高于宫颈上皮内瘤变组织,统计学有显著性差异(P<0.05)。
4.正常宫颈、宫颈上皮内瘤变与宫颈鳞癌组织中HIF-1αmRNA的含量没有显著性差异(P>0.05)。
5.高危型HPV DNA病毒载量在不同宫颈病变中差异无显著性。
6.锥切术后切缘阳患者持续或复发≥CIN2为20.63%(20/97)明显高于切缘阴患者的1.63%(9/551),p<0.01。锥切术后HC-Ⅱ结果持续阴性,复发例数为0,HC-Ⅱ结果持续阳性,复发率达25%(23/92)。
结论
1.L1蛋白在液基细胞涂片中的表达情况能够帮助了解宫颈病变的进展程度。HPVL1不表达或持续不表达者,提示病变进展。
2.宫颈癌前病变进展为宫颈癌时,14-3-3σ可能参与其过程。
3.hWAPL可能参与了宫颈上皮内瘤变的发生、发展、癌变的过程。
4.HR-HPV DNA病毒载量与宫颈病变的级别没有相关性,RLU/PC≥1即有临床意义,定性比定量更重要。
5.HC-Ⅱ检测对锥切术后随访有密切关系。
OBJECTIVES
1.To investigate whether it is possible to use detection of the human papillomavirus (HPV) L1 capsid protein to predict the course of mild or moderate cervical intraepithelial neoplasia(CIN).
2.To explore the expressions of 14-3-3σ,hWAPL and HIF-1αmRNA in cervical cancer and CINs。To analyze the role they play in the progression of CIN_s.
3.To observe the relation between cervical neoplasia and the viral load aquired by HCII. To assess the value of HC-Ⅱfor follow up after conization.
METHODS
1.Imrnunocytochemical analysis using antibody against HPV L1 capsid protein was carried out on 274 cases of TRIPATH Pap tests with abnormal cytologic diagnoses and positive for high-risk HPV DNA detected by HC-Ⅱ.
2.Collect fresh tissues from cervix.Extract total RNA and evaluate the relative expressions of 14-3-3σ,hWAPL and HIF-1αmRNA with real-time fluorogenetic quantitative PCR(RFQ-PCR).
3.A total of 2830 women were screened for cervical lesion.We assessed the relations between CINs and the viral load aquired by HC-Ⅱ.
4.648 patients accepted conization for≥CIN II histologically.We analyzed the changes of HC-Ⅱresults and the recurrences of lesions.
RESULTS
1.Cytologic diagnosis revealed a higher expression of L1 protein in LSIL(75.63%, 90/119)than in ASC-US(63.81%,67/105)and in HSIL/SCC(4/41,9.76%)(P<0.01).
2.71 cases without treatment accepted LCT test after lyear.In the cases expressing L1, 80%(44/55) regressed to NILM,and no one progressed.In the cases not expressing L1,only 37.50%(6/16) regressed to NILM,and 18.75%(3/16) progressed to ASC-H and HSIL.
3.The average quantity of 14-3-3σmRNA in cervical caner tissues was significantly higher than that in cervicitis tissues(t-test,P<0.05).
4.The average quantity of hWAPL mRNA in cervical caner and CINs tissues was significantly higher than that in cervicitis tissues(t-test,P<0.05).
5.The viral loads of high-risk HPV DNA in invasive cervical cancer and CINs were similar.
6.After conization,20.62%of the patients with positive surgical margins got persistent/recurrent lesions,while only 1.63%of those with negative surgical margins did.No persistent/recurrent lesions happened on the patients with persistent negative results of HC-Ⅱ.While 25%of those with persistent positive results of HC-Ⅱdid.
CONCLUSIONS
1.The expression of HPV L1 in LSIL was significantly higher than in HSIL.The disease may progress while not expressing L1 protein or non-expressing persistently.
2.14-3-3σand hWAPL may play an role in the emergence of cervical cancer.
3.The viral load of high-risk HPV DNA has no relation with the severity of cervical lesions.HC-Ⅱhas great significance for the follow-up after conization.
引文
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