Predictive Value of Marker of Proliferation Ki-67 and Cell Cycle Dependent Protein kinase Inhibitor P16INK4a in Cervical Biopsy to Determine Its Biological Behaviour

Objective: The aim of the study is to analyse the Immuno-histochemical expression of   Ki-67  and P16INK4a in CIN and cervical cancer cases  and  their utility to determine the accuracy of histological diagnosis and prediction of biological behavior of cervical lesion. Methodology: A retrospective cross-sectional study was carried in 110 numbers of cervical biopsy that included 25 CIN1, 21 CIN2, 12 CIN3, 26 SCC and 01 adenocarcinoma and 25 non neoplastic lesion. The tissue sections were stained with Ki-67 and P16INK4a. Results: Ki-67 expression was seen in 55.5% (61/110) cases of cervical lesion., out of which 3.6% (4/110; cervicitis -2/110 and metaplasia-2/110) cases were non dysplaia, 51.8% (57/110) cases were dysplasia /CIN of varying grade including invasive cancer. P16INK4a expression was noted 51.8% (57/110). There was an increasing trend of the intensity of Ki-67 and P16INK4a from focal positivity in low grade lesion to diffuse intensity in higher grade lesion and is statistically significant. There was strong association between the two variables Ki-67 and P16INK4a positive cases with their histologic grade. Conclusion: Though histopathology remains the ‘‘gold standard’’ for the diagnosis of CIN, both low and high-grade, biomarkers like Ki-67 and P16INK4a have emerged as helpful adjuncts. Their combined use may assist in the histopathologic classification of preinvasive lesions and facilitate the distinction from nondysplasia.     

Validation of p16INK4a as a marker of oncogenic human papillomavirus infection in cervical biopsies from a population-based cohort in Costa Rica.

Due to the high prevalence of cancer-associated types of human papillomavirus (HPV) and the poorly reproducible histologic classification of low-grade lesions, identifying infected women at highest risk for cancer prior to neoplastic progression remains a challenge. We therefore explored the utility of p16INK4a immunostaining as a potential diagnostic and prognostic biomarker for cervical neoplasia using paraffin-embedded tissue blocks (punch biopsies and loop electrosurgical excision procedures) obtained from women referred to colposcopy during the enrollment phase of the Guanacaste Project (1993 to 1994). All blocks from 292 women selected by HPV status (HPV negative, nononcogenic HPV positive, or oncogenic HPV positive) and representing the diagnostic spectrum of the population [normal to precancer: cervical intraepithelial neoplasia (CIN) 3] were immunostained for p16INK4a using the p16INK4a research kit based on the monoclonal antibody clone E6H4 (MTM Laboratories, Heidelberg, Germany). For CIN3, the sensitivity of diffuse p16INK4a immunostaining was 100% and the specificity was 95%. For CIN2, the sensitivity and specificity for diffuse staining were 81.1% and 95.4%, respectively. Generalized to the 10,000-woman cohort, this translated to positive predictive value and negative predictive value of 13.9% and 100% for CIN3, respectively, and 20.4% and 99.7% for CIN2 or CIN3, respectively. Of women with an initial diagnosis of less than CIN2 for whom follow-up data for up to 5 to 7 years were available, 44% with diffuse staining developed persistent infection (CIN2 or CIN3). Whereas our data support the diagnostic potential for p16INK4a, further prospective studies with detailed follow-up determining the prognostic capacity of this marker are needed.     

Predictive significance of the alterations of p16INK4A, p14ARF, p53, and proliferating cell nuclear antigen expression in the progression of cervical cancer.

PURPOSE: The purpose of this research was to evaluate the clinical significance of p16INK4A, p14ARF, p53, and proliferating cell nuclear antigen (PCNA) expression in tumor progression of cervical cancer. DESIGN: Seventeen patients (40 samples) with consecutive cervical lesions from normal squamous epithelium, inflammation of the cervix to cervical intraepithelial neoplasm (CIN) and invasive cervical squamous cell cancer (SCC), or from CIN to SCC were collected for this study. Expression of p16INK4A, p14ARF, p53, and PCNA were detected by immunohistochemistry on paraffin-embedded sections. Human papillomavirus DNA was detected simultaneously with PCR and typed according to its DNA sequence. RESULTS: p16INK4A overexpression was significantly higher in CIN (75%) and in SCC (75%) than in normal or inflammation of the cervix (12.5%; P < 0.01, P < 0.05, respectively). The positive rate of p14ARF expression was higher in SCC (83%) than in normal/inflammation of the cervix (25%; P < 0.05). PCNA expression was negative in normal or inflammation of the cervix, but an increased in expression was seen in 63.2% in CIN and 100% in SCC (P < 0.01, P < 0.05). When the time interval for disease progression from initial biopsy to CIN 3 or invasive cancer was compared with states of p16INK4A expression, cases stained positive for p16INK4A progressed within 64.2 months as compared with 122.3 months among those stained negatively (P < 0.01). Cases with increased p14ARF expression also had a short time interval for disease progression of 78.8 months as compared with 108.3 months in cases that were p14ARF negative. Cases with stable or decreased p53 expression had the shortest time interval for progression of 32.3 months in contrast to cases with no p53 expression (113.9 months). However, cases with increasing p53 expression progressed within 60.8 months. CONCLUSIONS: Our results suggested that altered states of p16INK4A, p14ARF, p53, and PCNA may be valuable markers to predict the progression of cervical neoplasia.     

P16INK4A、Brn-3a、C-myc及端粒酶在宫颈癌中的诊断价值研究

目的 探讨P16INK4A、Brn-3a及C-myc基因表达及端粒酶活性在宫颈癌诊断中的价值。方法 选取2012年3月至2014年5月接受手术治疗的宫颈癌患者40例,检测宫颈癌组织中P16INK4A、Brn-3a、C-myc的表达情况及端粒酶活性水平。同时选取40例宫颈上皮内瘤样病变（CIN）组织和28例正常宫颈组织作对比。采用受试者工作特征曲线（ROC）评价组织中P16INK4A、Brn-3a、原癌基因C-myc及端粒酶活性水平在宫颈癌诊断中的价值。结果 宫颈癌组织中P16INK4A、Brn-3a和C-myc的阳性表达率分别为100.0％、95.0％和100.0％,均高于正常宫颈和CIN组织,差异有统计学意义（P＜0.05）;CIN Ⅱ和CIN Ⅲ组织的P16INK4A阳性表达率高于正常宫颈和CINⅠ组织（P＜0.05）;正常宫颈组织中Brn-3a和C-myc阳性表达率均为7.14％,明显低于其他组织（P＜0.05）;宫颈癌组织中端粒酶活性水平为（44.38±3.82）U／g,高于其余各组,差异有统计学意义（P＜0.05）。Brn-3a、C-myc诊断宫颈癌的曲线下面积Az＞0.8,且灵敏度和特异度均在80.0％以上,而P16INK4A及端粒酶活性水平的诊断效能相对较低,但4个指标联合检测的诊断效能获提高,灵敏度和特异度分别为89.5％和83.4%,曲线下面积Az为0.879。结论 宫颈癌组织中P16INK4A、Brn-3a、C-myc表达水平和端粒酶活性异常,可能与宫颈癌的发生发展有密切关系,可作为宫颈癌诊断的参考指标。     

p16(INK4a) is a useful marker of human papillomavirus integration allowing risk stratification for cervical malignancies

The present study was conducted to assess utility of p16(INK4a) immunopositivity as a surrogate marker for genomic integration of high-risk human papillomavirus infection (hrHPV). A total of 29 formalin-fixed, paraffin-embedded cervical low-grade squamous intraepithelial lesions (LSILs), 27 high-grade squamous intraepithelial lesions (HSILs) and 53 invasive squamous cell carcinomas (SCCs), histologically-diagnosed between 1st January 2006 to 31st December 2008 at the University of Malaya Medical Centre were stained for p16(INK4a) (CINtec Histology Kit (REF 9511, mtm laboratories AG, Heidelberg, Germany). Immunopositvity was defined as diffuse staining of the squamous cell cytoplasm and or nucleus (involving > 75% of the intraepithelial lesions or SCCs). Staining of basal and parabasal layers of intraepithelial lesions was pre-requisite. One (3.4%) LSIL, 24 (88.9%) HSIL and 46 (86.8%) SCC were p16(INK4a) immunopositive. All normal squamous epithelium did not express p16(INK4). p16(INK4a) expression was significantly lower (p<0.05) in LSIL compared with HSIL and SCC with no difference in expression between HSIL and SCC.The increased p16(INK4a) immunopositivity in HSIL and SCC appears in line with the integrated existence of the hrHPV and may provide more insightful information on risk of malignant transformation of cervical squamous intraepithelial lesions than mere hrHPV detection.     

p16INK4a免疫细胞化学检测在筛查宫颈癌中的作用

目的 探讨p16^INK4a免疫细胞化学检测在筛查宫颈癌及其癌前病变中的作用。方法 选择220例宫颈液基细胞学剩余标本,制作液基薄片进行p16^INK4a 免疫细胞化学检测,随访组织活检结果,并与高危人乳头瘤病毒（HR—HPV）DNA检测结果进行对照。结果 p16^INK4a在宫颈细胞学诊断的鳞状细胞癌（SCC）、鳞状上皮内高度病变（HSIL）、鳞状上皮内低度病变（LSIL）、非典型鳞状细胞-小除外上皮内高度病变（ASC—H）和非典型鳞状细胞-不能明确意义（ASC—US）病例的阳性表达率分别为100．0％（7／7）、92．2％（107／116）、24．3％（17／70）、100．0％（14／14）和36．4％（4／11）。150例p16^INK4a阳性者中,111例有组织活检诊断,其中宫颈上皮内瘤变（CIN）2级及以上病变者97例（87．4％）;70例p16^INK4a阴性者中,18例有组织活检诊断,无一例CIN2及以上病变。p16^INK4a在CIN2及以上病变与在CIN1之间的阳性表达率差异有统计学意义（P〈0.01）,而HR-HPV DNA的阳性率在两者之间差异无统计学意义。结论 p16^INK4a在宫颈HSIL及以上病变中高表达,有利于高危病例的筛选。     

Evaluation of p16INK4a expression in ThinPrep cervical specimens with the CINtec p16INK4a assay: correlation with biopsy follow-up results

BACKGROUND: The aim of this study was to examine p16(INK4a) protein expression in ThinPrep (Cytyc Corporation, Marlborough, Mass) cervical specimens by using the CINtec p16(INK4a) Cytology Kit (Dako, Glostrup, Denmark). The ability of this assay to accurately identify underlying high-grade lesions was assessed by using follow-up biopsies and comparing these results with Hybrid Capture 2 (Digene, Gaithersburg, Md) high-risk HPV (hc(2)) results. METHODS: Three hundred ninety-eight residual ThinPrep samples were collected, and histological follow-up data were retrieved for abnormal cytology specimens. After preparation of a Papanicolaou-stained slide, a second slide was processed in preparation for p16(INK4a) immunostaining. High-risk human papillomavirus testing (hc(2)) was also performed. RESULTS: Of the 163 cytologically abnormal samples, 6-month biopsy follow-up data were available for 45% of the specimens. At initial blinded evaluation, 21 of the 26 cases with cervical intraepithelial neoplasia (CIN) II/III follow-up were positive for p16(INK4a), yielding an overall diagnostic sensitivity of 81%; 29 of the 47 cases diagnosed as CIN I or less were p16(INK4a) negative, yielding a diagnostic specificity of 62%. In comparison, the hc(2) test results indicated a diagnostic sensitivity of 100% with a diagnostic specificity of 15%. After review of selected cases with CIN II/III follow-up, 25 of 26 slides were deemed to be positive for p16(INK4a), increasing the diagnostic sensitivity to 96%. CONCLUSIONS: The CINtec p16(INK4a) Cytology Kit, in combination with ThinPrep cervical samples, allowed clear evaluation of p16(INK4a) protein overexpression. Diagnostic specificity of the CINtec p16(INK4a) assay was significantly improved relative to hc(2). To increase p16(INK4a) immunostaining in abnormal cells, a modified kit version with improved staining performance has been developed and is currently being evaluated.     

Comparison of p16INK4a Immunocytochemistry with the HPV Polymerase Chain Reaction in Predicting High Grade Cervical Squamous Intraepithelial Lesions

Aim: To compare p16INK4a immunocytochemistry with the HPV polymerase chain reaction in predictinghigh grade cervical squamous intraepithelial lesions. Materials and Methods: This diagnostic case-control studywas conducted from January 2010 until December 2010. We obtained 30 samples, classified according to thedegree of cervical intraepithelial neoplasia (CIN): 11 samples for CIN 1, 9 samples for CIN 2, and 10 samples forCIN 3. HPV PCR, p16INK4a immunocytochemistry, and histopathological examination were performed on allsamples. Statistical analysis was conducted using SPSS 20.0. Results: In predicting CIN 2-3, we found p16INK4ato have similar specificity and positive predictive value as HPV PCR (95%, 97.2% vs 96.7%), but better sensitivity(87.5% vs 72.5%) and negative predictive value (82.1% vs 67.6%). The most prevalent types of high-risk HPVin our study were HPV 33, 35, 58, 52, and 16. Conclusions: p16INK4a has better diagnostic values than HPVPCR and may be incorporated in the triage of ASCUS and LSIL to replace HPV PCR. Genotype distribution ofHPV differs in each region, providing a challenge to develop HPV vaccines based on the epidemiology of HPVin that particular region.     

乳头瘤病毒L1衣壳蛋白及p16INK4A基因在子宫颈病变中的表达及其相关性

 目的　研究乳头瘤病毒衣壳蛋白（HPVL1）及p16INK4A基因在子宫颈病变患者中的表达与相关性。方法　采用免疫组织化学Max-Vision法检测210例HPV阳性液基细胞学标本及组织中HPV L1蛋白及p16INK4A的表达。结果　在各级液基细胞学标本中,HPVL1蛋白阳性率各组比较差异有统计学意义（χ2＝70.50、P＜0.005）,其中低度鳞状上皮内病变（LSIL）组最高[68 %（34／50）];p16INK4A表达阳性率随着病变级别的增高逐渐增高（13 %、28 %、52 %、100 %、100 %）,在子宫颈癌（SCC）组最高[100 %（30／30）]。HPVL1+p16INK4A -率在LSIL组中最高[32 %（16／50）];HPVL1-p16INK4A +率在SCC组最高[100 %（30／30）]。在各级别病理组织中,HPVL1蛋白表达阳性率不同,差异有统计学意义（χ2＝54.37、P＜0.005）,其中,CINⅠ级中最高[60.4 %（32／53）];p16INK4A表达阳性率随着病变级别的增高逐渐增高,在子宫颈癌组最高[100 %（28／28）]。HPVL1+p16-率在CINⅠ级中最高[45.3 %（24／53）];HPVL1-p16+率在子宫颈癌中最高[100 %（28／28）]。结论　HPVL1蛋白与p16INK4A联合检测可以区分不同的子宫颈病变,将有潜在发展与有自愈可能的病例分开,避免漏诊与过度治疗。     

Combined p16INK4a and human papillomavirus testing improves the prediction of cervical intraepithelial neoplasia (CIN II-III) in Thai patients with low-grade cytological abnormalities

Thailand is in the process of developing a national cervical screening program. This study examined p16INK4a staining and HPV prevalence in abnormal cervical samples with atypical squamous cells of undetermined significance (ASCUS) and low-grade squamous intraepithelial lesion (LSIL), to evaluate the efficacy of combined HPV and p16INK4a detection to predict CIN II-III. Totals of 125 ASCUS and 87 LSIL cases were re-evaluated by Pap test and cervical cells of ASCUS and LSIL cases were prepared on slides for p16INK4a detection by immunocytochemistry. HPV genotyping of DNA extracts was performed by GP5+/6+ PCR and reverse line blot hybridization. Histopathologic tests were performed to identify cervical lesion. Total of 212 cases were diagnosed to normal (20), ASCUS (112), LSIL (78) and HSIL (2). HPV was detected in ASCUS (49/112, 43.8%), LSIL (60/78, 76.9%) and HSIL (2/2, 100%) cases. The majority of HPV positive samples typed for high-risk HPV. 55.7% (107/192) of abnormal cases (ASCUS, LSIL and HSIL) were positive p16INK4a. For the 111 HPV DNA positive cases, 34 of 49 (69.4%) ASCUS cases and 49 of 60 (81.7%) LSIL cases were p16INK4a positive. 140 biopsies were taken and histological classified: CIN negative (65 cases), CIN I (56 cases) and CIN II-III (19 cases). HPV DNA detection predicted CIN II-III with sensitivity and specificity of 84% and 49%, whereas p16INK4a staining showed higher sensitivity (89.5%) and specificity (56.2%). The prediction of CIN II-III was significantly better by combination of positive HPV DNA and p16INK4a with 93.8% sensitivity and 59.2% specificity. Detection of HPV DNA combined with p16INK4a in cervical cells can predict CIN II-III and may improve the screening diagnosis of Thai women at risk for CIN II-III or cancer.     

P16INK4a和Ki-67在宫颈癌前病变诊断中的意义

目的研究P16INK4a、Ki-67在宫颈癌前病变中的表达及早期诊断价值。方法宫颈活检标本92例,采用免疫组织化学PV-9000二步法检测宫颈鳞癌28例,CIN 45例（CINⅢ级17例、CINⅡ级7例、CINⅠ级21例）,炎性病变19例。结果在宫颈癌、CIN病变、宫颈炎P16INK4a的阳性表达率分别为100%、44.4%和10.5%,Ki-67的阳性表达率分别为100%、51.11%、21.05%,三组不同病变比较均差异有显著性（P〈0.001）。P16INK4a与Ki-67表达存在正相关（r2=0.893,P〈0.001）。结论 P16INk4a、Ki-67联合检测可以提高宫颈癌的早期诊断率。     

P16INK4a及Ki-67免疫细胞化学检测对ASC-US中高级别病变检出的意义

目的：探讨液基细胞学剩余细胞标本P16INK4a、Ki-67免疫细胞化学染色检测宫颈癌前病变的价值。方法：选取2010年1月至2012年6月因宫颈疾病于北京大学第三医院妇产科就诊患者液基细胞学剩余标本50例。所有患者行高危型HPVDNA杂交捕获II代（HCII）检测,P16INK4a、Ki-67免疫细胞化学检测,同时行阴道镜检查及组织病理学活检。结果：以病理诊断将患者分为CIN2以下组和CIN2及以上组。CIN2及以上组P16INK4a及Ki-67表达量高于CIN2以下组,差异均具有统计学意义（P〈0．05）;P16INK4a或Ki-67检测对高级别病变预测的准确性在ASC—US组中优于异常细胞学组;在ASC—US中,P16INK4a或Ki-67检测对高级别病变预测的准确性优于高危型HPV检测。结论：宫颈脱落细胞中P16INK4a或Ki-67免疫细胞化学检测相比于高危型HPV检测可以提高对ASC—US中高级别病变的检出作用。     

p16和Ki-67免疫组化检测与宫颈上皮内瘤变(CIN)程度的相关性分析

目的:分析免疫组化检测p16和Ki-67与宫颈上皮内瘤变（CIN）严重程度的关系,以及 p16联合Ki-67在高级别CIN鉴别中的效能。方法:采用回顾性研究,纳入2016年1月至2018年3月期间粤北人民医院经HE染色病理确诊的宫颈病变患者160例,其中慢性宫颈炎者共38例归为对照组,CINⅠ级、Ⅱ级、Ⅲ级分别为41例、46例、35例;再将CIN患者分为低级别鳞状上皮内病变（LSIL）和高级别鳞状上皮内病变（HSIL）组。对比对照组与CINⅠ级、Ⅱ级、Ⅲ级组、ISIL组、HSIL组p16及Ki-67阳性表达情况;采用Spearman相关性分析观察p16阳性表达、Ki-67阳性表达与CIN严重程度的关系。联合使用p16、Ki-67半定量评分,评估联合p16、Ki-67半定量评分诊断结果与病理结果一致性,评估效能。结果:CINⅠ-Ⅲ级组p16阳性率较对照组高,CINⅡ、Ⅲ级p16阳性率较CINⅠ级高;HSIL组阳性率较LSIL组高,差异均有统计学意义,P＜0.05。Ki-67在对照组中阳性率为0.00%,在LSIL组、HSIL组中分别为68.29%、100.00%。p16与宫颈病变程度呈正相关,r=0.692,P=0.021 0;Ki-67与宫颈病变程度呈正相关,r=0.612,P=0.001 8。p16联合Ki-67诊断炎症结果与病理完全一致,CINⅠ级共36例（87.80%）符合,CINⅡ级共43例（93.48%）符合,CINⅢ级共34例（97.14%）符合;LSIL符合率为87.80%（36/41）,HSIL符合率为95.06%（77/81）。p16联合Ki-67诊断LSIL敏感性、特异性为94.74%和97.58%,诊断HSIL敏感性、特异性分别为96.25%、98.81%,诊断CIN敏感性、特异性分别为92.62%、96.48%,其中LSIL敏感性高于单用,CIN诊断特异性较单用p16高,P＜0.05。结论:p16和Ki-67表达与宫颈上皮内瘤样病变严重程度密切相关,可以通过观察二者表达情况辅助CIN病情严重程度判断,联合p16和Ki-67免疫组化检测能提高诊断高级别鳞状上皮内病变诊断特异性、敏感性。     

p16INK4A表达与宫颈癌患者临床病理特征及预后的关系

目的 探讨p16INK4A(p16)蛋白在宫颈癌和宫颈上皮内瘤变(CIN)中的表达及其与临床病理特征和预后的相关性.方法 选取宫颈活检和手术切除的宫颈癌患者的宫颈标本100例,CIN组织80例以及正常宫颈组织50例,采用免疫组织化学法对p16蛋白的表达进行检测,并对所得结果进行统计分析.结果 p16在宫颈癌组织、CIN组织与正常宫颈组织中的阳性表达率比较,差异有统计学意义(P<0.05);p16蛋白的表达强度和阳性表达率与宫颈病变程度呈正相关(r=0.545,P<0.05);p16蛋白阳性表达与宫颈癌的分化程度及淋巴结转移情况有关(P<0.05);p16阳性表达患者的3年生存率低于p16阴性表达患者(P<0.05).结论 p16蛋白表达与宫颈癌的发生、发展及不良预后有关,可以作为判断宫颈癌生物学行为和预后评价的指标.     

ｐ１６和Ｋｉ-６７蛋白在宫颈上皮内瘤变中的临床意义

目的　研究ｐ１６和Ｋｉ-６７蛋白在宫颈组织中的表达,探讨它们在宫颈病变中的诊断价值。方法　应用免疫组化技术ＳＰ法检测正常宫颈或炎性病变组织２０例、ＣＩＮⅠ组织２２例、ＣＩＮⅡ组织２０例、ＣＩＮⅢ组织２０例、宫颈腺体受累及组织２０例（未见原发病灶）、宫颈鳞状细胞癌组织２０例中ｐ１６和Ｋｉ-６７蛋白的表达情况。结果　ｐ１６和Ｋｉ-６７蛋白在正常宫颈组织中的阳性表达率明显低于宫颈癌前病变及宫颈癌组织,差异有显著意义（Ｐ＜０.０１）。Ｋｉ-６７蛋白阳性表达在良性宫颈组织与ＣＩＮⅠ组织相比较,无显著差异（Ｐ＞０.０５）;ｐ１６阳性表达在宫颈基底细胞增生组织与ＣＩＮⅠ组织相比较,有显著差异（Ｐ＜０.０５）,而Ｋｉ-６７蛋白无显著差异（Ｐ＞０.０５）;ｐ１６着色强阳性在ＣＩＮⅠ与ＣＩＮⅡ、ＣＩＮⅢ、宫颈癌比较,有显著差异（Ｐ＜０.０１）。结论　ｐ１６蛋白能很好地鉴别宫颈良性病变、宫颈上皮内瘤变以及宫颈癌;在鉴别宫颈基底细胞增生与ＣＩＮⅠ时,ｐ１６能起辅助诊断作用;应用ｐ１６的着色强弱能够很好地区分ＣＩＮⅠ与ＣＩＮⅡ、ＣＩＮⅢ,有助于ＣＩＮ的判断与分级。Ｋｉ-６７能很好地区分宫颈高度病变与宫颈癌,但不能区分宫颈良性病变与宫颈低度病变,需结合ｐ１６蛋白联合判断。     

Evaluation of cervical cone biopsies for coexpression of p16INK4a and Ki-67 in epithelial cells

Diffuse overexpression of p16(INK4a) in basal and parabasal cells of cervical epithelium is a hallmark of human papillomavirus-mediated transformation. Focal p16(INK4a) expression is occasionally observed in nondysplastic epithelium. In normal cells, expression of p16(INK4a) triggers cell cycle arrest. However, cells undergoing transformation in intraepithelial lesions actively proliferate. To prove that the different expression patterns of p16(INK4a) , i.e., focal versus diffuse, reflect biologically different entities, we hypothesized that p16(INK4a) -positive cells in epithelia displaying focal p16(INK4a) expression pattern do not coexpress proliferation-associated Ki-67 protein, while p16(INK4a) -positive cells in lesions with diffuse p16(INK4a) expression may do. A total of 138 cervical cone biopsies were stained for the expression of p16(INK4a) and Ki-67 using a primary antibody cocktail. All metaplastic lesions (n = 21) displayed focal staining for p16(INK4a) , and in all of these lesions p16(INK4a) -positive cells were found to be negative for Ki-67 expression. Diffuse expression of p16(INK4a) was observed in 12/21 (57.1%) cervical intraepithelial neoplasia (CIN) 1 lesions, all of them simultaneously showed Ki-67 immunoreactivity in a large proportion of p16(INK4a) -positive cells. Seventeen of 23 (73.9%) CIN2 lesions and all 27 (100%) CIN3/carcinoma in situ (CIS) as well as all 46 (100%) carcinoma cases displayed diffuse and combined expression of p16(INK4a) and Ki-67. Coexpression of Ki-67 and p16(INK4a) in the same cell is entirely restricted to cervical lesions displaying diffuse p16(INK4a) expression, whereas in lesions with focal p16(INK4a) expression, p16(INK4a) -expressing cells are negative for Ki-67. Thus, diffuse expression of p16(INK4a) reflects lesions with proliferation-competent cells, while p16(INK4a) -expressing cells associated with focal expression patterns are cell cycle arrested.     

Human papillomavirus testing for triage of women with low‐grade squamous intraepithelial lesions

Low‐grade squamous intraepithelial lesion (LSIL) is a common cytologic finding in cervical screening, yet only about 10–20% have significant histologic abnormalities and these are almost always positive for high‐risk human papillomavirus (hrHPV). This analysis aims to clarify the role of hrHPV DNA testing in the triage of women with LSIL cytology. In the ATHENA screening trial, we examined 1,084 cases of LSIL, of which 925 had an evaluable biopsy, to determine the extent to which hrHPV testing can identify those patients who have precursor lesions in need of immediate clinical referral and those who have changes more likely to regress spontaneously. Overall, 71.2% of LSIL cases were hrHPV positive, but the prevalence was age dependent, with only 56.1% in women ≥40 years. Among women with LSIL, 11.6% (107/925) had a cervical intraepithelial neoplasia grade 2 or worse (CIN2+) histologic diagnosis and, of these, only nine were hrHPV negative. For CIN3+, 91.7% (44/48) of women with LSIL were hrHPV positive. The negative predictive value of hrHPV testing for CIN3+ in LSIL was 100% for women aged ≥40 years. Women who were HPV16 positive had a higher positive predictive value for CIN2+ (25.4%) than those who were positive for 12 other pooled hrHPV types (11.5%). Testing for hrHPV in women with LSIL is effective in identifying high‐grade cervical lesions, thereby avoiding unnecessary referrals to colposcopy and potential over‐treatment of non‐progressive lesions, especially for women aged ≥40 years.     

318例高级别宫颈上皮内瘤变及宫颈癌患者的TCT和hrHPV检测分析

  目的  探讨如何使用现有的筛查方法, 减少高级别宫颈上皮内瘤变(cervical intraepithelial neoplasia grade 2 or 3, CIN2/3)及宫颈癌的漏诊。  方法  回顾性分析2014年6月至2018年8月318例中国医科大学附属航空总医院治疗的CIN2/3及宫颈癌患者的临床资料, 其中CIN2/3为296例、宫颈癌为22例, 采用宫颈液基薄层细胞学检查(thinprep cytology test, TCT)及高危型人乳头瘤病毒(high risk human papilloma virus, hrHPV)检测方法, 分析患者的年龄、TCT和hrHPV。  结果  296例CIN2/3患者中30~39岁患者为130例(43.92%)、占第1位, 20~29岁年轻患者为69例(23.31%)、占第3位。318例患者中TCT阳性为199例(62.58%), hrHPV阳性为308例(96.86%), 两者联合筛查阳性为313例, 阳性率为98.43%(313/318)。hrHPV分型检测主要亚型依次为16、52、58、33、18、31型。  结论  CIN2/3及宫颈癌的发病年龄年轻化, 年轻患者筛查不容忽视。TCT单独筛查较hrHPV单独筛查易漏诊高级别病变, TCT联合hrHPV筛查可提高检出率。TCT阴性, hrHPV非16、18亚型的其他hrHPV阳性, 尤其是hrHPV52、58、33、31亚型阳性患者也建议行阴道镜检查。