hTERC基因异常扩增在宫颈病变的临床意义

目的探讨应用FISH技术检测人类染色体端粒酶（hTERC）基因异常扩增在子宫颈病变的临床意义。方法选择经过液基细胞学（TCT）检查的130例患者为研究对象,用荧光原位杂交（FISH）方法检测脱落细胞hTERC基因异常扩增情况,与病理结果金标准相比对。结果130例患者中正常细胞学妇女20例,病理检查炎症26例,C IN1患者34例,C IN2/3患者34例,鳞状细胞癌（SCC）患者16例,FISH检测hTERC基因异常扩增阳性率在炎症组是3.8%、C IN1组23.5%、C IN2/3组82.4%和SCC组100%,炎症组和C IN1组与C IN2/3组和SCC组比较,hTERC基因扩增阳性率差异有显著统计学意义（P〈0.005）。随着病变程度增加,hTERC基因异常扩增阳性率增加。hTERC基因检测高级别病变（C IN2/3、鳞状细胞癌）的敏感度、特异度分别是88.0%、88.8%。C IN组hTERC基因平均拷贝数为2.18,SCC组平均拷贝数为3.05,SCC组hTERC基因拷贝数比C IN组明显增加（P〈0.01）,FISH检测结果显示,杂交信号hTERC基因高拷贝型比例在炎症组6.1%,C IN1组17.0%,C IN2/3组33.8%,SCC组47.7%,呈上升趋势（P〈0.05）。宫颈病变向高级别进展组（n=7）与病变维持低级别或低级别转归组（n=12）的hTERC基因异常扩增阳性比分别为5/7和0/12,两组有明显差异（P〈0.01）。平均拷贝数分别为2.19和2.04,两组有明显差异（P〈0.05）。结论hTERC基因的异常扩增随宫颈病变级别增高而增加,并对宫颈早期病变进展有预测意义。应用FISH技术检测hTERC基因的异常扩增可作为监测宫颈病变进展的生物遗传学指标,协助TCT和高危HPV DNA检测诊断高级别C IN和鳞状细胞癌。     

高危型人乳头瘤病毒感染的子宫颈非典型鳞状细胞中p16基因的缺失

目的：观察p16基因在高危型人乳头瘤病毒（ human papi11omavirus,HPV）感染的子宫颈非典型鳞状细胞（ atypi-ca1 squamous ce11s of undermined significance,ASCUS）中的缺失,探讨其作为子宫颈病变筛查的临床意义。方法收集27例高危型HPV-DNA检测为阳性、液基薄层细胞学（ thin-prep cyto1ogic test,TCT）诊断为ASCUS的脱落细胞标本。27例患者均在1个月内进行阴道镜检查并行组织学活检,根据组织学结果分组：对照组（包括炎症和HPV感染）7例;子宫颈上皮内瘤变（ cervica1 intraepithe1ia1 neop1asia,CIN ）组20例,其中CIN13例、CIN213例、CIN34例。制作细胞蜡块并切片,采用FISH技术检测ASCUS中p16基因缺失情况。结果 p16基因在对照组和CIN组中的缺失率分别为14.3%和85.0%,二者差异有统计学意义（ P<0.01）。结论采用FISH技术检测ASCUS细胞中p16基因缺失有助于从伴有ASCUS患者中筛选出子宫颈CIN患者。     

高危人乳头瘤病毒在宫颈癌筛查中的应用及卫生经济学评价

据2002年数据统计,全球估计有49万的子宫颈癌新发病例,超过27万妇女死于该病。目前高危人乳头瘤病毒（high risk-human pappiloma virus,HR-HPV）联合细胞学检测是大多数发达国家和发展中国家经济发达地区妇女首选的子宫颈癌筛查方案。研究显示,HPV检测可明显提高宫颈高度病变的检出率,提高细胞学检查的敏感性,从卫生经济学角度来看,两者联合检测是一个低本高效的方案,HR-HPV与细胞联合检测是目前宫颈癌筛查的最佳选择,但仍存在一定的局限性。     

人乳头瘤病毒感染与宫颈病变的相关性

目的 探讨宫颈病变与人乳头瘤病毒感染的情况.方法 对我院2008年5月～2009年12月妇科门诊和住院患者528人次,同时通过HPV-DNA检测及宫颈液基细胞学和病理检测结果,同时通过阴道镜下多点位活检病理对比来诊断CIN患者528人次,对其HPV感染情况和CIN病变程度之间的关系进行分析.结果 感染CIN1患者有193例占36.7%,HPV感染率84.3%,CIN2-3中有患者335例占63.3%.HPV感染率92.4%,经过分析统计其HPV感染率和CIN病情程度有统计学意义(P<0.05).结论 CIN感染等级程度与HPV病毒感染有相关性.     

hTERC和HPV联合检测在宫颈上皮内瘤变筛查中应用

目的 观察人端粒酶mRNA基因(human telomerase mRNA componentgene,hTERC)在宫颈上皮内瘤变(cervical intraepithelial neoplasia,CIN)及宫颈癌中的表达情况,探讨其作为宫颈病变筛查的临床意义.方法 收集135例宫颈疾病患者的宫颈活检组织标本.根据组织学结果分组,其中正常组(包括炎症患者)24例;CIN组100例,其中CIN 1级30例,CIN 2级50例,CIN 3级20例;宫颈癌(鳞癌)组11例.采用双色间期FISH技术检测宫颈细胞hTERC基因扩增情况,同时其中的85例患者进行了HPV-DNA检测,阳性者49例,阴性者36例.结果 (1)hTERC基因在正常组、CIN组及宫颈癌组中的阳性表达率分别为4.2%、38.0%和90.9%,各组间差异有统计学意义(P<0.01),与病变程度呈正相关(r=0.419,P<0.01).(2)hTERC基因在CIN 1～3级组中的阳性表达率分别为10.0%、48.0%和55.0%,CIN 2级组与CIN 3级组比较差异无统计学意义(P>0.05),CIN 1～3级组间比较差异有统计学意义(P<0.01),与病变的恶性程度呈正相关(r=0.354,P<0.01).(3)49例HPV阳性患者中,hTERC基因扩增阳性率为57.14%(28/49);而在36例HPV阴性患者中,hTERC基因扩增阳性率为25.0%(9/36),两者差异比较有统计学意义(P<0.005),同时hTERC基因的扩增情况与HPV感染情况两者间呈正相关(r=0.320,P<0.003).结论 hTERC基因的阳性率随宫颈病变级别的升高而增加,可预测病变的进展情况.hTERC基因的扩增和HPV感染呈正相关,提示HPV的感染可能是hTERC基因异常扩增的早期事件.HPV-DNA和hTERC基因联合检测相结合是宫颈CIN早期筛查的理想手段.     

宫颈上皮内瘤样病变患者高危型HPV感染基因分型分析

目的了解宫颈上皮内瘤变患者的高危型HPV的感染及其分型和不同程度宫颈病变的主要感染型别情况。方法应用型特异PCR检测宫颈癌前病变的患者的主要高危型HPV-16、18、33、58的感染及其分型情况的相关性研究。结果在本研究宫颈上皮内瘤变患者98例中,4种高危型HPV的总阳性例数为73例,HPV总的感染率为74.5%,存在多重感染。其中HPV-16、18、33、58的总感染率分别为53.1%、38.7%、17.3%和30.6%。CIN的Ⅰ/Ⅱ/Ⅲ3组患者的4种高危型HPV的感染率分别为42.9%、61.1%和93.2%。结论主要高危型HPV在宫颈上皮内瘤变患者中感染的主要型别依次为HPV16、HPV18、HPV58、HPV33,主要为HPV16和HPV18型;不同程度CIN的高危型HPV的总感染率不同,随病变程度的加重而增加。     

子宫颈刮片中人乳头瘤病毒的基因分型

目的 确定不同型人乳头瘤病毒（ＨＰＶ）感染的自然历程以及其持续感染在子宫颈癌发展过程中的作用。方法 应用聚合酶链反应检测荷兰１５５名妇女子宫刮颈片中的ＨＰＶ ＤＮＡ，应用线样探针分析法（ＬｉＰＡ）进行，包括ＨＰＶ６，１１，１６，１８，３１，３３，３５，４０，４２，４３，４４，４５，５１，５２，５６和５８的基因分型。结果 １５５例妇女子宫颈片中ＨＰＶ ＤＮＡ检出率为６０％，其中在宫颈细胞学检查正常或     

人端粒酶RNA基因荧光原位杂交检测在宫颈细胞学检查中的应用

目的 探讨人类染色体端粒酶RNA基因(hTERC)扩增的荧光原位杂交(FISH)检测在宫颈脱落细胞防癌筛查中的意义.方法 收集2008年2-10月上海长海医院门诊就诊146例患者宫颈液基细胞学样本,应用间期双色FISH技术检测hTERC基因的扩增情况,并与细胞学和组织学结果进行对照.结果 杂交成功120例(细胞学阴性20例、细胞学阳性100例).hTERC基因扩增的阳性率与细胞学等级成正相关(r=0.465,P＜0.01),与组织学等级成正相关(r=0.610,P＜0.01),各级的阳性率分别为:炎性病变0(0/13)、宫颈上皮内瘤样病变(CIN)Ⅰ级2 8.6%(6/21)、CINⅡ级11/18、CINⅢ级75.0%(18/24)、鳞状细胞癌91.7%(22/24);FISH方法检出高级别(CINⅡ/Ⅲ级)以上病变的敏感性为77.3%(51/66),特异性为82.4%(28/34).阳性预测值89.5%,阴性预测值65.1%,低级别(CIN Ⅰ级)以下病变的hTERC阳性率与高级别以上病变相比差异有统计学意义(x2=32.550,P＜0.01).结合hTERC高倍扩增(信号数＞4)的出现,检出高级别以上病变的敏感性提高到81.2%.结论 hTERC基因扩增的FISH检测有助于辅助细胞学诊断,提高高危病变的检出率.实验结果判断除参照阈值之外,还应结合hTERC基因的扩增类型,出现高倍扩增亦提示高级别以上病变.

Abstract：

Objective To investigate the value of fluorescence in situ hybridization (FISH)detection of human telomerase RNA component ( hTERC ) gene amplification in screening of cervical lesions.Methods A total of 146 post-thinPrep cytology test (TCT) samples were analyzed using FISH by two-color interphase probe targeting hTERC gene at chromosome 3q26 and the data were compared with the cytological and histological results. Results FISH analysis was successful in 120 cases (20 cases of normal and 100 abnormal cases by TCT). Gene amplification of hTERC by FISH had a positive correlation with the cytological (r = 0. 465, P ＜ 0. 01 ) and histological grade results ( r = 0. 610, P ＜ 0. 01 ). Extra copies of hTERC were seen in 28.6% (6/21) of CIN Ⅰ , 61.1% (11/18) of CIN Ⅱ , 75.0% (18/24) of CIN Ⅲ and 91.7% (22/24) of squamous cell carcinoma, respectively. None (0/13) of the inflammation cases showed hTERC amplification. The sensitivity and specificity for detecting high grade lesions by FISH were 77. 3%(51/66) and 82. 4% (28/34) ;and the positive and negative predictive values were 89. 5% and 65. 1%,respectively. The rate of hTERC gene gain in high grade lesions was significantly higher than that in the low grade lesions( x2 =32. 550,P ＜0. 01 ). Combined with the high copy numbers, the sensitivity for detecting high grade lesions was increased to 81.2%. Conclusions Detection of hTERC gene amplification by FISH improves the screening efficiency of high-risk cervical epithelial lesions. The presence of high copy numbers of hTERC correlates with the presence of high grade cervical dysplasia.     

TERC基因检测在不同级别宫颈病变中的诊断价值

目的检测不同级别宫颈病变中TERC基因的表达情况,探索其在不同宫颈病变中的诊断价值。方法采用荧光原位杂交（fluorescence in situ hybridization,FISH）技术检测20例正常对照和100例患者（CINⅠ14例、CINⅡ35例、CINⅢ36例、宫颈癌15例）的宫颈脱落上皮细胞TERC基因的表达情况。并以组织病理学结果做对照。结果随宫颈病变级别增加,TERC基因扩增的阳性率、异常细胞数和异常核型的复杂性均显著增加。宫颈癌／CINⅢ该基因扩增阳性率显著高于CINⅡ／Ⅰ及正常对照（P〈0．01）,CINⅡ病变者显著高于CINⅠ者（P〈0．01）。结论TERC基因扩增与宫颈病变的发展关系密切,FISH检测TERC基因扩增在不同级别宫颈病变的诊断中有重要意义。     

人乳头瘤病毒阳性宫颈癌和宫颈上皮内瘤变患者血浆中miR-34a的表达及临床意义

目的 探究miR-34a在人乳头瘤病毒(HPV)阳性宫颈癌和宫颈上皮内瘤变(CIN)患者血浆中的表达及其对HPV阳性宫颈癌的诊断价值。方法 选取2016年1月至2021年1月期间在本院就诊的HPV阳性宫颈癌患者80例作为宫颈癌组,并选取同一时间与宫颈癌组患者年龄无差异的HPV阳性CIN患者60例作为CIN组,HPV阳性但子宫良性病变者50例作为对照组。通过qRT-PCR检测3组受试者血浆中miR-34a的表达情况,并分析其与HPV阳性宫颈癌患者临床病理特征的关系;检测血浆中CA125和SCCA的表达情况;使用Pearson法分析HPV阳性宫颈癌患者血浆中miR-34a与CA125或SCCA水平的相关性;经ROC曲线评估miR-34a与CA125、SCCA对HPV阳性宫颈癌的诊断价值。结果 宫颈癌组患者血浆中miR-34a表达水平明显低于CIN组和对照组(0.48±0.15vs 0.74±0.28和0.96±0.38,F=50.796,P<0.05),且CIN组低于对照组(P<0.05)。宫颈癌组患者血浆中CA125(U/mL)、SCCA(ng/mL)水平明显高于CIN组和对...     

Cervical intraepithelial neoplasia and aneusomy of TERC: Assessment of liquid‐based cytological preparations

The implementation of effective screening programs has decreased the incidence and mortality of cervical carcinoma. However, single screening tests are subjective and carry a significant false‐negative rate. Therefore, supplementary tests to support the Papanicolaou (PAP) smear are being developed. Human papillomavirus (HPV) testing has increased the specificity of the PAP smear, but has high sensitivity rate. This has proven unhelpful in low‐grade lesions and in young women. Cervical carcinogenesis is a multifactorial disorder. In addition to exposure to oncogenic HPV, which is regarded as the initiator, there must be a promoter to eventuate in invasive disease. The promoter factor appears to be the acquisition of extra copies of chromosome 3q and has been shown to be a constant recurrence in cervical carcinoma (squamous and adenocarcinomas). The 3q region contains the RNA sequence of the human telomerase gene TERC. Recent studies have shown a strong correlation between high‐grade cervical lesions and abnormalities of TERC. This study supports the previous studies and examines the status of the TERC gene in low and high‐grade cervical intraepithelial lesions, diagnosed on cytology. ASC‐H smears are also examined in an attempt to categorize lesions that are more likely to progress. Potentially this may help identify women in need of close clinical follow‐up and early treatment. Diagn. Cytopathol. 2009. © 2009 Wiley‐Liss, Inc.     

High Grade Cervical Intraepithelial Neoplasia and Viral Load of High-Risk Human Papillomavirus: Significant Correlations in Patients of 22 Years Old or Younger

High-risk human papillomavirus (HR-HPV) is recognized as the primary cause for the development of cervical cancers and their precursor lesions. We investigated whether high-grade cervical dysplasia correlates with high viral load of HR-HPV in an age-dependent manner. Cases were retrospectively selected to include patients with a prior cytological diagnosis of ASCUS or higher grade squamous intraepithelial lesions, and a positive Digene Hybrid Capture II (HC-II) HR-HPV testing within 2 months before or after cervical biopsy. The quantitative viral load data was classified as negative, low, moderate and high according to the manufacturer’s instruction. Cases were then stratified into 4 age groups: ≤22 years, 23-30 years, 31-40 years and >40 years. Chi-Square analysis and logistic regression were performed where appropriate. A total of 995 patients were identified. Age categories were significantly associated with HPV loads (p=0.046). Moderate to high viral loads of HPV were significantly related to the histological grade of dysplasia (p=0.029). Logistic regression analysis further confirmed the association of HPV with histological grades, even after adjusting for age factor. In particular, high-grade dysplasia (p=0.011) but not low grade dysplasia (p=0.140) was significantly associated with moderate to high HPV loads. Patients of 22 years old or younger were the only group found significantly correlated with high viral loads of HPV (p=0.015). In conclusion, high-grade squamous intraepithelial lesions and patients’ age of 22 years old or younger are significantly associated with a moderate to high viral load of HR-HPV.     

FISH技术检测TERC基因在子宫颈上皮内瘤变1级与2级鉴别诊断中的实用价值

目的：采用FISH技术检测TERC基因在子宫颈上皮内瘤变( cervical intraepithelial neoplasia, CIN)不能明确1级或2级(CIN1/2)病变组织中的扩增,探讨其在CIN分级中的临床病理实用意义。方法选取子宫颈组织标本42例,其中正常子宫颈鳞状上皮20例,CIN1/2级22例(其中初步诊断1级8例,2级14例)。采用FISH技术检测石蜡包埋组织中TERC基因的扩增情况。结果正常子宫颈鳞状上皮中TERC基因无扩增,CIN1/2级组织中TERC基因的扩增率为22．7%(5/22),77．3%(17/22)的TERC基因无扩增;其中初步诊断1级8例,1例TERC基因扩增;初步诊断2级14例,4例TERC基因扩增,差异有统计学意义(P<0．05)。结论在石蜡包埋组织切片难以确定CIN1/2级时,应用FISH技术进行TERC基因检测,TERC基因扩增者可考虑为CIN2级,建议临床积极处理;无扩增者,考虑为CIN1级,可保守处理;FISH技术具有临床病理实用价值。     

Different amplification patterns of the human telomerase RNA gene in invasive cervical carcinomas and cervical intraepithelial neoplasia grade III

The aim of this study was to compare the amplification patterns of the human telomerase RNA gene (hTERC) in invasive cervical carcinomas (ICC) and cervical intraepithelial neoplasia grade III (CIN III) and to define their potential clinical implications. Cervical liquid-based cytological (LBC) specimens were collected from 53 squamous cell carcinomas (SCC), 14 CIN III, and 20 normal controls. Copy numbers of the hTERC gene were measured by fluorescence in situ hybridization (FISH) using a dual-color probe containing the hTERC probe and the control, chromosome 3 centromere-specific probe (CSP3). Nucleus with abnormal FISH pattern for hTERC was observed in 0.94-90.65% of SCC cells and in 0-85.59% of CIN III cells. Using the threshold of 5.89%, the occurrence of hTERC amplification in SCC and CIN III was similar (90.6% vs. 85.7%, P = 0.630). However, the median percentage of cells with extra gains of hTERC (hTERC:CSP3 > 1) in SCC was higher than in CIN III (64.3% vs. 31.7%, P = 0.001). Among those cells, the 3:2 signal pattern was the leading pattern for both SCC and CIN III; high-level amplification of hTERC was more common in SCC than in CIN III (60.9% vs. 48.9%, P = 0.002). In SCC, it was not found that extra gains of hTERC were associated with any clinicopathological parameters. Thus, hTERC amplification was common in cervical exfoliated cells from SCC and CIN III. More complex amplification patterns of hTERC were present in ICC. Clinical usefulness of hTERC amplification in LBC samples was limited in ICC.     

HPV感染与宫颈癌前病变

宫颈癌是一个由癌前病变逐渐衍变为癌的连续的病理过程。目前认为宫颈癌前病变,即宫颈上皮内瘤变与HPV感染有关。HPVs是一种双链小DNA病毒,由病毒蛋白外壳和核心DNA物质构成。病毒基因组分为早期基因区、晚期基因区及长调控区;其中早期区编码的E6、E7蛋白对于病毒的复制起关键作用。生殖道HPVs在有性活动的人群中普遍存在,在有性生活的女性中,至少有75%将在人生中的某个时间感染HPV。感染HPV后绝大多数人可以自然消退。只有感染了高危亚型、同时又具备其他高危因素的妇女才可能进展为HSIL或宫颈癌。因此对于HPV的感染既要重视,又不必恐惧。目前的治疗主要是针对由HPV引起的宫颈或外生殖器的局部病变。     

Silver staining method for nucleolar organizer regions in cervical smears

The distribution of nucleolar organizer regions (NORs) was studied in Papanicolaou preparations of cervical smears in order to distinguish benign from preneoplastic lesions. Destained smears (six defined as normal, six as inflammatory with squamous metaplasia, six as CIN I, six as CIN II, and five as CIN III) were submitted to the Ag-NOR method after staining with Orange G and EA36. Ag-NOR count was performed in previously outlined fields on the smears. Statistically significant differences (P < .05) were found between the normal smears, inflammatory smears with squamous metaplasia, and each grade of CIN. We conclude that the Ag-NOR technique could be useful to evaluate cervical smears of doubtful interpretation, using previous demarcation of the abnormal fields/cells. Diagn. Cytopathol. 16:497–499, 1997. © 1997 Wiley-Liss, Inc.     

Clinicopathological Implications of Human Papilloma Virus (HPV) L1 Capsid Protein Immunoreactivity in HPV16-Positive Cervical Cytology

Background: The objective of this study was to investigate the expression of human papilloma virus (HPV) L1 capsid protein in abnormal cervical cytology with HPV16 infection and analyze its association with cervical histopathology in Korean women.Material and Methods: We performed immunocytochemistry for HPV L1 in 475 abnormal cervical cytology samples from patients with HPV16 infections using the Cytoactiv^® HPV L1 screening set. We investigated the expression of HPV L1 in cervical cytology samples and compared it with the results of histopathological examination of surgical specimens.Results: Of a total of 475 cases, 188 (39.6%) were immunocytochemically positive and 287 (60.4%) negative for HPV L1. The immunocytochemical expression rates of HPV L1 in atypical squamous cells of unknown significance (ASCUS), low-grade squamous intraepithelial lesions (LSIL), high-grade squamous intraepithelial lesions (HSIL), and cancer were 21.8%, 59.7%, 19.1%, and 0.0%, respectively. LSIL exhibited the highest rate of HPV L1 positivity. Of a total of 475 cases, the multiple-type HPV infection rate, including HPV16, in HPV L1-negative cytology samples was 27.5%, which was significantly higher than that in HPV L1-positive cytology samples (p = 0.037). The absence of HPV L1 expression in ASCUS and LSIL was significantly associated with high-grade (≥cervical intraepithelial neoplasia [CIN] 2) than low-grade (≤CIN1) histopathology diagnoses (p < 0.05), but was not significantly different between HPV16 single and multiple-type HPV infections (p > 0.05). On the other hand, among 188 HPV L1-positive cases, 30.6% of multiple-type HPV infections showed high-grade histopathology diagnoses (≥CIN3), significantly higher than the percentage of HPV16 single infections (8.6%) (p = 0.0004)Conclusions: Our study demonstrates that the expression of HPV L1 is low in advanced dysplasia. Furthermore, the absence of HPV L1 in HPV16-positive low-grade cytology (i.e., ASCUS and LSIL) is strongly associated with high-grade histopathology diagnoses. The multiplicity of HPV infections may have an important role in high-grade histopathology diagnoses (≥CIN3) in HPV L1-positive cases.     

Comparative genomic hybridization as a tool in tumour cytogenetics

The quality of cytogenetic analysis of solid tumours has greatly improved in the past decade, but a number of technical difficulties remain which limit the characterization of solid tumour chromosomes by conventional cytogenetics alone. The identification of regions of chromosomal abnormality has been aided by the introduction of molecular cytogenetic techniques such as fluorescence in situ hybridization (FISH). Of these, a recently developed approach, comparative genomic hybridization (CGH), has had a particular impact on the cytogenetic analysis of solid tumours. It incorporates the sensitivity of in situ techniques and overcomes many of the drawbacks of conventional cytogenetic analysis. This review first outlines the CGH method, giving details for the preparation of DNA probes and target human metaphase chromosomes together with information on the in situ technique and data handling criteria used in our laboratory. It then presents an overview of some of the current applications of CGH, together with a discussion of future directions in the field. Copyright © 1999 John Wiley & Sons, Ltd.