p16、Ki-67在宫颈鳞状上皮病变分级诊断中的意义

目的:探讨p16、Ki-67在宫颈鳞状上皮病变分级诊断中的意义.方法:收集我院2013年6月至2015年6月,阴道镜检查并宫颈活检病理127例,进行免疫组化p16、Ki-67检测,整理完整临床和病理资料作为研究对象.结果:根据127例宫颈活检HE染色及p16、Ki-67免疫组化结果修订原诊断.26例诊断为慢性宫颈炎;32例HSIL除1例考虑萎缩性改变,31例诊断HSIL;58例LSIL其中7例诊断为HSIL;1 1例CIN1-2中2例诊断为HSIL,9例诊断为LSIL;所有修订后HSIL,均随访1年.40例HSIL患者行Leep、宫颈锥切治疗,病理诊断除1例表现为p16、Ki-67阴性外(考虑萎缩性改变),与活检病理诊断一致.结论:联合免疫组化p16、Ki-67检测,在宫颈鳞状上皮病变分级诊断、治疗中具有重要意义,p16在肿瘤性病变中过表达更具特异性.     

p16、Ki-67在宫颈鳞状上皮病变分级诊断中的意义

目的:探讨p16、Ki-67联合应用在宫颈鳞状上皮病变分级诊断中的意义.方法:采用免疫组化方法检测117例宫颈各类病变活检组织中p16、Ki-67蛋白表达情况,并对该组患者液基薄层细胞学(TCT)、高危型人乳头瘤病毒基因检测(HPV-DNA)结果进行比较.结果:根据HE组织学形态,结合免疫组化染色结果修订原有诊断,其中LSIL、HSIL的构成比差异有统计学意义.p16在LSIL、HSIL中的阳性表达率分别为0.00%、97.30%,Ki-67则分别为21.57%、67.58%(P<0.05).TCT和HPV-DNA检测HSIL的敏感度89.19%,特异度39.61%.联合应用p16、Ki-67的敏感度46.67%,特异度95.74%.结论:联合免疫组化染色检测p16、Ki-67可作为宫颈鳞状上皮病变分级诊断的重要标记物.联合TCT、HPV-DNA在宫颈癌筛查中有较高的敏感度,但特异度有限,需与p16、Ki-67免疫组化染色相结合.     

宫颈上皮内瘤变p16（INK4a）和p53及Ki-67表达及其诊断价值探讨

目的：探讨用p16和p53及Ki-67等免疫组化标记作为宫颈上皮内瘤变（CIN）诊断的辅助指标。方法：选取我院档案CIN患者133例,年龄20～86岁,中位年龄46岁。经3位高年资病理医师复查确认,其中CIN 1级31例、CIN 2级37例和CIN 3级65例。另选同期我院宫颈慢性炎患者19例,浸润性鳞状细胞癌患者20例作为对照。采用SP法进行p16、p53和Ki-67免疫组化染色,结果独立评分。各级别CIN与免疫组化表达的关系采用单变量χ2检验和Spearman等级相关分析。结果：p16阳性表达位于细胞核或胞核伴胞质,p53和Ki-67定位于细胞核。p16、p53和Ki-67表达阳性率随着CIN的加重而升高,均与CIN级别呈正相关,r值分别为0.789、0.554和0.749,P〈0.001。153例CIN 1＋标本中p16、p53和Ki-67与组织学诊断符合率分别为96.7%（148/153）、71.9%（110/153）和88.2%（135/153）。在102例CIN 2＋标本中,p16、p53和Ki-67免疫组化染色的敏感性分别为97.1%（99/102）、74.5%（76/102）和97.1%（99/102）;特异性分别为94.7%（18/19）、84.2%（16/19）和73.7%（14/19）。p16、p53和Ki-67 3者联合在CIN 2＋中的敏感性和特异性分别为98.0%（100/102）、68.4%（13/19）。结论：在宫颈活检的组织学评价时,结合p16、p53和Ki-67免疫组化染色,对提高高级别CIN的检出率、减少漏诊有很大作用。     

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。     

宫颈上皮内瘤样病变发生发展与p16/Ki-67共表达的相关性研究进展

宫颈上皮内瘤样病变（cervical intraepithelial neoplasia,CIN）属于宫颈癌前病变,虽然CIN进展成宫颈癌时间一般较长,但其发病率增多及发病年龄变小,对女性身心健康造成严重影响。随着宫颈癌筛查技术的进步和分子生物学技术的发展,高级别的宫颈癌上皮内瘤样变的早期检查确认并进行阻断已成为筛查的目标。p16被认为是发现最早的抑癌基因,在鉴别低级别与高级别宫颈鳞状上皮内病变中有重要作用。Ki-67为常见肿瘤标志物,在多种肿瘤中表达。p16/Ki-67双染为新型宫颈病变筛查或诊断方法,即同一细胞内检测到p16、Ki-67共表达,则提示细胞周期失调,考虑为宫颈上皮内高级别瘤样病变。国内近年有研究报道p16和Ki-67在CIN中的作用,未见到相关的综述报道。本文对宫颈上皮内瘤样病变与p16/Ki-67共表达的相关性作一综述,为临床宫颈病变鉴别、诊断提供参考。     

宫颈液基细胞学标本中p16^INK4A的表达

[目的]探讨p16^INK4A在新柏氏宫颈液基细胞学（TCT）标本中各级宫颈上皮内瘤变的表达，评价其在宫颈病变中表达的意义。[方法]采用Envision法，检测76例TCT标本及相应宫颈活检组织中p16^INK4A表达。[结果]在TCT中，P16^INK4A在无上皮内病变（NILM）、非典型鳞状细胞（ASC）、低级别鳞状上皮内病变（LSIL）、高级别鳞状上皮内病变（HSIL）阳性表达率分别为0、15．78％、45．45％、88％。在相应活检组织中，p16^INK4A在宫颈炎、CINI、CINⅡ、CINⅢ中阳性表达率分别为0、63．15％、89．28％、100％。p160^INK4A在TCT和相应活检组织中表达的符合率为91.23％。[结论]TCT标本的p16^INK4A表达检测可以提高宫颈高级别病变的检出率。     

阴道镜联合p16/Ki-67双染对不典型鳞状细胞诊断的应用价值

[目的]探究阴道镜联合p16/Ki-67双染对不典型鳞状细胞(ASC-US)诊断中的应用价值。[方法]选取2016年1月到2019年1月410例经液基细胞学检测确诊为ASC-US患者,患者均行阴道镜和组织病理检查,取其细胞学样本进行p16/Ki-67双染检测。[结果]宫颈上皮瘤变2级及以上有(CIN2+)161例,其中2级72例,3级82例,鳞癌9例。阴道镜检测发现CIN2+的敏感度、特异性、诊断符合率分别为82.0%、64.3%和71.2%;p16/Ki-67双染检测发现CIN2+的敏感度、特异性、诊断符合率分别为77.6%、77.1%和68.9%;阴道镜检查、p16/Ki-67双染检测及联合诊断CIN2+的ROC曲线下面积分别为0.731、0.744和0.860。[结论]阴道镜联合p16/Ki-67双染检测具有较高的敏感度和特异性,准确率高于单一检测方法,能较准确分流ASC-US。     

宫颈非典型性不成熟鳞状上皮化生的特征及诊断

目的探讨宫颈非典型性不成熟鳞状上皮化生（AIM）的特征及在诊断和鉴别诊断中的意义。方法对55例原诊断为宫颈CINⅢ级的病例和宫颈息肉上皮成熟性鳞状化生（OSM）14例，采用病理形态学观察，AB粘液组织化学染色，Ki-67免疫组织化学染色，半巢式PCR方法检测HPV16DNA的方法，观察组织病理学特征，粘液残留情况，细胞增殖活性及HPV病毒感染情况。结果根据鳞状上皮中的粘液残留情况，从55例CINⅢ级中鉴别出7例AIM：AIM的Ki-67表达高于OSM（P=0．022），明显低于CINⅢ级（P=0．000）：AIM与高危型HPV病毒感染相关。结论AIM可能是一类与高危型HPV感染有关的低级别的鳞状上皮内病变，有着与宫颈高级别鳞状上皮内瘤变相似的组织形态特征。本研究认为对AIM的诊断和鉴别诊断要参考其组织形态特征，粘液残留现象，细胞增殖指数和HPV的感染情况。     

宫颈鳞状上皮病变组织中乳头状瘤病毒和Ki-67及p16^INK4a蛋白表达意义的研究

目的：探讨Ki-67、p16^INK4a和人乳头状瘤病毒（HPV）在不同程度宫颈鳞状上皮病变组织中的表达及临床病理意义。方法：采用原位分子杂交及免疫组化方法,检测HPV的不同亚型、Ki-67、p16^INK4a蛋白在182例不同程度宫颈病变组织中的表达。结果：HPV在不同程度病变中总检出率52.19%（95/182）;在宫颈高级别上皮内瘤变及鳞癌组中检出最多的感染类型为HPV16/18,而HPV6/11在尖锐湿疣组检出率87.50%（21/24）最高;随着宫颈病变严重程度的增加,级别升高,Ki-67、p16^INK4a阳性程度呈递增趋势。Ki-67、p16^INK4a与HPV16/18型感染关系密切,χ^2=11.779 8,P〈0.01;Ki-67也与HPV6/11型有关。结论：HPV16/18型及Ki-67、p16^INK4a在宫颈高级别上皮内瘤变及鳞癌中表达明显升高,可能对宫颈鳞癌的发生、发展具有协同作用。     

P16/Ki-67蛋白双染在未明确意义的非典型鳞状上皮细胞人群中分流作用

摘 要：［目的］探讨p16/Ki-67免疫细胞双染法对宫颈细胞学为未明确意义的非典型鳞状上皮细胞（ASCUS）人群的分流效果。［方法］ 以2016年4月至12月在郑州大学第二附属医院妇科门诊就诊且被诊断为ASCUS的135例妇女为研究对象,收集其宫颈脱落细胞标本,进行14种高危型HPV DNA及p16/Ki-67蛋白检测,所有妇女均进行阴道镜活检和病理学检查。以组织病理学诊断为金标准,分别计算p16/Ki-67双染、高危型HPV（HR-HPV）和HPV16/18检测的敏感性、特异性、阳性预测值（PPV）、阴性预测值（NPV）、转诊率及其95%CI。［结果］ 135例ASCUS妇女平均年龄为（46.48±10.19）岁（23~64岁）,其中诊断为宫颈上皮内瘤变（CIN）1级者7例,诊断为CIN2级及以上（CIN2+）者22例。随异常病理结果的严重性增加,p16/Ki-67双染、HR-HPV和HPV16/18检出阳性率均升高（P<0.001）。以CIN2+作为疾病终点指标时,p16/Ki-67双染对ASCUS人群分流的敏感性、特异性、PPV、NPV和转诊率分别为86.4%(95%CI：66.7%~95.3%)、85.8%(95%CI：78.2%~91.1%)、54.3%(95%CI：38.2%~69.5%)、97.0%(95%CI：91.6%~99.0%)和25.9%,与之相比,HR-HPV检测敏感性稍高［95.5%(95%CI：78.2%~ 99.2%)］,但特异性较低［68.1%(95%CI：59.1%~76.0%)］,转诊率较高（42.2%）,且差异均有统计学意义（P<0.05）。与p16/Ki-67双染法相比,HPV16/18的特异性较高［92.9%(95%CI：86.7%~96.4%)］,但敏感性很低［59.1%(95%CI：38.7%~76.7%)］。按照45岁进行年龄分层后,p16/Ki-67双染在≥45岁组ASCUS人群中分流效果要更好,敏感性为81.8%(95%CI：52.3%~94.9%),特异性为95.5%(95%CI：87.5%~98.4%)。［结论］ P16/Ki-67双染检测在保持高敏感性的同时,有更高的特异性,因此对ASCUS人群的分流效果优于HR-HPV和HPV16/18。P16/Ki-67双染检测具有简便、客观、高效、易于重复的特点,可为ASCUS人群提供一种新的分流方法。     

P16/Ki-67 Dual Staining in Positive Human Papillomavirus DNA Testing for Predictive Diagnosis of Abnormal Cervical Lesions in Northeastern Thai Women

Objective: Cervical cancer screening can effectively reduce new cervical cancer cases, including in Thailand. The abnormal results are subsequently referred for colposcopy. To avoid unnecessary colposcopy, an efficient triage is still needed for validation. This study aimed to investigate the overall positivity of cytology-based screening, HPV detection, and p16/Ki-67 dual staining and evaluate different triage strategies for predictive diagnosis of abnormal cervical lesions in northeastern Thailand. Methods: Cervical cells were collected from 191 women who came for cervical screening in the gynecological outpatient department during March 2019-February 2020. Pap smear samples were classified into 6 groups including 17 atypical glandular cells (AGC), 21 atypical squamous cells of undetermined significance (ASC-US), 7 atypical squamous cells - cannot exclude HSIL (ASC-H), 26 low-grade squamous intraepithelial lesions (LSILs), 19 high-grade SILs (HSILs) and 101 no squamous intraepithelial lesion (noSIL). Polymerase chain reaction (PCR) was performed for HPV DNA detection. HPV genotyping was determined by reverse line blot hybridization. P16/Ki-67 dual staining was performed by using CINtec PLUS Cytology kit. Biopsies from abnormal screening were collected for surgical pathology classification. Results: High-risk HPV (HR-HPV) infection was 2.97%, 29.41%, 38.10%, 57.14%, 46.15% and 84.21% in noSIL, AGC, ASC-US, ASC-H, LSIL and HSIL cytology respectively. P16/ Ki-67 in noSIL, AGC, ASC-US, ASC-H, LSIL and HSIL was 0.99%, 5.88%, 9.52%, 42.86%, 26.92% and 63.16%, respectively (P-value < 0.001). Among p16/Ki-67 positive cases, 96.15% (25/26) were infected with HPV and 84.62% (22/26) were HR-HPV. The overall positivity of each and co-testing between cytology or HPV DNA testing or p16/Ki-67 dual staining was evaluated. In each cervical lesion, primary HPV DNA testing showed the highest sensitivity, but low specificity. The combined all HPV/HR-HPV with p16/Ki-67 detection increased the specificity of abnormal cervical lesions. Conclusion: P16/Ki-67 dual stain cytology in HPV-positive women performs well for diagnosis of abnormal cervical lesions and should be considered for management of HPV-positive women to avoid unnecessary colposcopy referrals.     

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.     

Geminin、Ki-67和HPV检测对早期宫颈上皮内瘤变的诊断价值

目的:探讨Geminin、Ki-67和HPV检测对早期宫颈上皮内瘤变的诊断价值。方法:采用免疫组化SP法检测Geminin、Ki-67在126例宫颈上皮内瘤变(CIN)组织、50例慢性宫颈炎宫颈组织及30例正常宫颈组织中的表达；采用HC2的方法检测HPV的载量。用Spearman秩相关分析Geminin、Ki-67分别与HPV感染的相关性。结果:Geminin、Ki-67在CIN组织中阳性表达率高于宫颈炎组和对照组(P＜0.05)；Geminin、Ki-67在宫颈炎组和对照组中阳性表达率差异无统计学意义(P＞0.05)；Geminin、Ki-67在CIN组织中阳性表达率CINIII组＞CINII组＞CINI组，组间比较差异有统计学意义(P＜0.05)；HPV在宫颈病变组织中阳性表达率CINIII组＞CINII组＞CINI组＞宫颈炎组＞对照组，组间比较差异有统计学意义(P＜0.05)；Geminin、Ki-67表达分别与HPV感染呈正相关(P＜0.05)；Geminin、Ki-67和HPV联合检测的特异度和精确度最高。 结论:不同CIN分期病变组织Geminin、Ki-67表达水平和HPV感染率差异较大，可通过HPV、Geminin、Ki-67联合检测对早期CIN进行辅助诊断。     

Diagnostic Performance of p16/Ki-67 Dual Immunostaining at Different Number of Positive Cells in Cervical Smears in Women Referred for Colposcopy

BackgroundThe aim of the study was to evaluate the diagnostic accuracy of p16/Ki-67 dual immunostaining (p16/ Ki-67 DS) in cervical cytology and the number of positive p16/Ki-67 cells to diagnose high grade cervical intraepithelial neoplasia (CIN2+) in colposcopy population.Subjects and methodsWe performed an analysis on a subset cohort of 174 women enrolled within a large-scale randomised controlled human papillomavirus (HPV) self-sampling project organised as part of the population-based Cervical Cancer Screening Programme ZORA in Slovenia. This subset cohort of patients was invited to the colposcopy clinic, underwent p16/Ki-67 DS cervical cytology and had the number of p16/Ki-67 positive cells determined.ResultsAmong analysed women, 42/174 (24.1%) had histologically confirmed CIN2+. The risk for CIN2+ was increasing with the number of positive cells (p < 0.001). The sensitivity of p16/Ki-67 DS for detection of CIN2+ was 88.1%, specificity was 65.2%, positive predictive value was 44.6% and negative predictive value was 94.5%.ConclusionsDual p16/Ki-67 immunostaining for the detection of CIN2+ has shown high sensitivity and high negative predictive value in our study, which is comparable to available published data. The number of p16/Ki-67 positive cells was significantly associated with the probability of CIN2+ detection. We observed a statistically significant and clinically relevant increase in specificity if the cut-off for a positive test was shifted from one cell to three cells.Key words: cervical cytology, high-grade dysplasia, p16/Ki-67 immunostaining     

Combined use of cytology,p16 immunostaining and genotyping for triage of women positive for high-risk human papillomavirus at primary screening

Human papillomavirus (HPV) testing is very sensitive for primary cervical screening but has low specificity. Triage tests that improve specificity but maintain high sensitivity are needed. Women enrolled in the experimental arm of Phase 2 of the New Technologies for Cervical Cancer randomized controlled cervical screening trial were tested for high-risk HPV (hrHPV) and referred to colposcopy if positive. hrHPV-positive women also had HPV genotyping (by polymerase chain reaction with GP5+/GP6+ primers and reverse line blotting), immunostaining for p16 overexpression and cytology. We computed sensitivity, specificity and positive predictive value (PPV) for different combinations of tests and determined potential hierarchical ordering of triage tests. A number of 1,091 HPV-positive women had valid tests for cytology, p16 and genotyping. Ninety-two of them had cervical intraepithelial neoplasia grade 2+ (CIN2+) histology and 40 of them had CIN grade 3+ (CIN3+) histology. The PPV for CIN2+ was >10% in hrHPV-positive women with positive high-grade squamous intraepithelial lesion (61.3%), positive low-grade squamous intraepithelial lesion (LSIL+) (18.3%) and positive atypical squamous cells of undetermined significance (14.8%) cytology, p16 positive (16.7%) and, hierarchically, for infections by HPV33, 16, 35, 59, 31 and 52 (in decreasing order). Referral of women positive for either p16 or LSIL+ cytology had 97.8% sensitivity for CIN2+ and women negative for both of these had a 3-year CIN3+ risk of 0.2%. Similar results were seen for women being either p16 or HPV16/33 positive. hrHPV-positive women who were negative for p16 and cytology (LSIL threshold) had a very low CIN3+ rate in the following 3 years. Recalling them after that interval and referring those positive for either test to immediate colposcopy seem to be an efficient triage strategy. The same applies to p16 and HPV16.     

子宫颈鳞状细胞癌中人乳头瘤病毒感染与Rb相关基因蛋白的表达及其意义

 目的　探讨子宫颈浸润性鳞状细胞癌（ISCC）中高危型人乳头瘤病毒（HPV）16／18、31／33感染与Rb相关基因蛋白p16ink4a、CyclinD1、CDK4、Rb、E2F-1和Ki-67的表达情况并分析HPV及Rb相关蛋白与ISCC临床病理参数的相关性。方法　收集73例ISCC、30例高级别鳞状上皮内病变（HSIL）、32例低级别鳞状上皮内病变（LSIL）和21例正常子宫颈鳞状上皮（NCE）。采用原位杂交法检测HPV16／18、31／33DNA的表达;免疫组织化学二步法检测上述蛋白的表达。统计分析HPV、上述蛋白及临床病理参数的关系。结果　HPV16／18、31／33在四组总体间表达差异有统计学意义（P＜0.01）,HPV16／18阳性表达与脉管内瘤栓正相关（OR＝3.875）,HPV31／33阳性表达与临床分期正相关（OR＝3.5）;p16ink4a、CyclinD1、CDK4、Rb、E2F-1和Ki-67在四组总体间差异有统计学意义（P＜0.01）。Rb与Ki-67正相关（rs=0.250）;p16ink4a阳性表达与组织分级负相关（OR＝0.942）;CDK4阳性表达与组织分级正相关（OR＝1.033）;年龄与临床分期正相关（OR＝1.063）。结论　HPV16／18、31／33感染与ISCC的发生、发展密切相关;CDK4、Ki-67可作为子宫颈病变程度的客观参考指标;Rb在ISCC中的详细作用机制还需进一步探讨。