p16、p53和Ki-67蛋白在宫颈上皮内瘤变中的表达及意义

目的 探讨p16、p53和Ki-67蛋白在宫颈上皮内瘤变(cervical intraepithelial neoplasia,CIN)中的表达及临床意义.方法 采用免疫组化SP法检测正常子宫颈或炎性病变组织、CIN1～3中p16、p53和Ki-67蛋白的表达.结果 p16、p53和Ki-67蛋白在正常子宫颈或炎性病变中罕见表达,在CIN1～3组织中三者表达均较高,随CIN级别升高p16、p53和Ki-67表达增强,各组间表达差异有统计学意义(P<0.05).同时p16、p53和Ki-67三者阳性表达均可见分层现象,在CIN1中大部分阳性细胞位于子宫颈鳞状上皮的下1/3,在CIN2中多累及上皮下2/3,而CIN3则普遍超过上皮的下2/3或全层弥漫阳性,各组间差异具有统计学意义(P<0.05).结论 p16、p53和Ki-67蛋白表达均与子宫颈上皮内瘤变的病变进展密切相关,联合检测p16和Ki-67的抗原表达可作为CIN分级诊断的辅助方法,具有较好的应用价值.     

P16、Ki-67和Bcl-2在宫颈上皮内瘤变中的表达及应用

目的探讨P16、Ki-67、Bcl-2抗体在宫颈上皮内瘤变(CIN)中的表达及应用价值。方法用快捷免疫组化染色法检测30例正常宫颈组织和66例CIN中p16、Ki-67和Bcl-2的表达,分析比较两组的标记结果。结果 CIN中p16、Ki-67、Bcl-2的阳性率显著高于正常宫颈(﹤0.05),CINⅡ-Ⅲ的表达率显著高于CINⅠ(﹤0.05)。结论 P16、Ki-67和BCL-2抗体联合标记可作为病理诊断CIN和区分级别的重要参考指标。     

p16INK4a在宫颈细胞学鳞状上皮内瘤变中的意义

目的探讨p16^INK4a的表达在宫颈细胞学中鳞状上皮内瘤变中的意义及其与人乳头状瘤病毒（HPV）型别之间的关系。方法对88例经活检证实的液基细胞学标本[包括20例慢性宫颈炎、18例低度鳞状上皮内瘤变（LSIL）、34例高度鳞状上皮内瘤变（HSIL）及16例鳞状上皮细胞癌（SCC）],分别用免疫细胞化学（EnVision方法）检测其p16^INK4a的表达,并对所有标本用聚合酶链反应（PCR）方法检测HPV型别（包括HPV16、18、31、33、35、39、45、51、52、53、56、58、59、66、68、6、11、42、43及44型）。结果p16^INK4a在慢性宫颈炎组呈阴性,在LSIL、HSIL及SCC组的表达率分别为27．8％、100％及100％,LSIL、HSIL及SCC组p16^INK4a的表达均显著高于慢性宫颈炎组（P〈0．01）;p16^INK4a在高危型HPV感染组的表达率（96．4％）显著高于低危型HPV感染组（7．7％）,差异有统计学意义（t=4．32,P〈0．01）。结论p16^INK4a是一种敏感性较高,特异性较好的标记物,可以识别与高危型HPV有关的非典型鳞状上皮细胞。     

宫颈鳞状上皮内病变与EBP50、p16和Ki-67表达的相关性分析

目的:评价Ezrin-radixin-moesin结合磷酸化蛋白50(Ezrin-radixin-moesin binding phosphoprotein 50,EBP50)、p16和Ki-67在正常宫颈组织和宫颈鳞状上皮内病变(Squamous intraepithelial lesion,SIL)中的表达及临床意义.方法:选取2018年10月至2020年06月正常宫颈组织21例、SIL组织80例,采用免疫组化方法检查EBP50、p16和Ki-67表达水平,通过相关性检验分析EBP50与p16、Ki-67表达的相关性.结果:EBP50在正常宫颈组织中表达明显高于SIL(P=0.000);SIL中p16和Ki-67表达显著高于正常宫颈组织中的表达(P=0.000);高级别鳞状上皮内病变(High-grade squamous intraepithelial lesion,HSIL)EBP50表达强度比低级别鳞状上皮内病变(Low-grade squamous intraepithelial lesion,LSIL)降低(P=0.003);EBP50与p16或Ki-67的表达在SIL中呈负相关.结论:EBP50、p16和Ki-67的检测对SIL的病理诊断具有参考价值.EBP50表达的降低与宫颈病变的级别升高相关,EBP50可作为判断SIL分级的有效辅助手段.     

胃肠道间质瘤中p16、p27、Ki-67表达

目的探讨胃肠道间质瘤中p16、p27和Ki67表达与临床预后的关系。方法根据核分裂象的多少、肿瘤体积的大小及有无浸润等将胃肠道间质瘤划分为良性、交界性和恶性,并运用免疫组化SP方法检测p16、p27和Ki67在胃肠道间质瘤中的表达,并统计分析其良性、交界性、恶性和复发死亡病例中的表达差异。结果p16、p27和Ki67的阳性表达率分别为48%、26%和24%。p16在良、恶性中的表达无明显差异,但在良性和交界性中的表达与复发和转移相关;p27低标记指数和Ki67高标记指数与复发和转移相关。结论p16、p27和Ki67在胃肠道间质瘤中的表达对判断预后有价值。     

新疆维吾尔族妇女宫颈上皮内瘤变及宫颈癌中FHIT、P16~(INK4a)、RARβ蛋白的表达及意义

目的探讨脆性组氨酸三联体(fragile histidine triad,FHIT)、P16INK4a与视黄酸受体β(retinoic acid receptor-beta,RARβ)蛋白在新疆维吾尔族妇女宫颈上皮内瘤变(cervical intra-epithelial neoplasia,CIN)及宫颈癌中的表达及其意义。方法采用免疫组化链霉素抗生物素蛋白-过氧化物酶链接(SP)法检测20例慢性宫颈INK4a炎、30例CIN(CINⅠ、CINⅡ、CINⅢ各10例)以及40例浸润性宫颈鳞癌组织标本中FHIT、P16INK4a及RARβ蛋白的表达。结果(1)FHIT蛋白阳性表达率依次为慢性宫颈炎(90.00%)CIN(66.67%)浸润性宫颈鳞癌INK4a(27.50%)(P=0.000);P16和RARβ蛋白阳性表达率依次为浸润性宫颈鳞癌(82.50%;90.00%)CIN(46.67%;53.33%)慢性宫颈炎(0.00%;10.00%)(P=0.000;P=0.000)。(2)在宫颈各组织中,FHIT蛋白INK4a与P16INK4a蛋白表达呈负相关(r=-0.384,P=0.000);FHIT蛋白与RARβ蛋白表达呈负相关(r=-0.291,P=0.006);P16INK4a蛋白与RARβ蛋白表达呈正相关(r=0.445,P=0.000)。结论抑癌基因FHIT表达缺失与P16INK4a和RARβ过度表达与宫颈癌的发生发展密切相关,并且FHIT、P16INK4a、RARβ具有协同作用。FHIT蛋白与P16INK4a蛋白和RARβ蛋白的联合检测可作为宫颈癌早期诊断和宫颈癌进展的分子指标。     

子宫颈病变中p16INK4A 蛋白表达及其与HPV16/18感染的关系

目的 探讨p16INK4A 蛋白在子宫颈鳞癌(SCC)和子宫颈上皮内肿瘤(CIN)中的表达及其与HPV16/18感染的关系.方法 用原位杂交法检测HPV16/18在25例子宫颈癌、45例CIN及10例慢性子宫颈炎中的表达,同时用免疫组化EliVision法检测p16INK4A 蛋白的表达.结果 (1)与慢性子宫颈炎相比,CIN Ⅱ级、CIN Ⅲ级、浸润癌HPV16/18杂交信号阳性率显著增高(P<0.01);(2)子宫颈鳞癌组织、CIN Ⅰ级、CIN Ⅱ级、Ⅲ级及慢性子官颈炎标本中p16INK4A 蛋白阳性率分别为100.0%、20.0%、46.7%、100.0%和10.0%;(3)在子宫颈鳞癌及CIN HPV16/18感染的标本中p16INK4A 蛋白表达均是阳性.结论 子宫颈鳞癌的形成与HPV感染、p16INK4A 蛋白过表达是呈正相关关系,p16INK4A蛋白可能作为子宫颈鳞癌及CIN的标志物,对子宫颈癌筛查和预防有重要意义.     

绝经后妇女子宫颈鳞状上皮中p16与Ki-67的表达和意义

目的比较绝经后妇女子宫颈高级别鳞状上皮内病变(high-grade squamous intraepithelial lesion,HSIL)与萎缩性改变对p16与Ki-67的免疫组化表达差异,探讨两者的鉴别诊断。方法选取绝经后妇女诊断为HSIL的病例19例,诊断为萎缩性改变的病例26例,采用免疫组化ELPS法标记p16与Ki-67两种抗体,观察此两种抗体阳性细胞表达的部位,染色强度与染色分布。结果HSIL组对p16与Ki-67两种标记物均呈不同程度的阳性表达。对于p16,17例(89.4%)阳性细胞呈全层分布,各有1例(5.3%)阳性细胞出现在鳞状上皮的上2/3或下2/3。14例(73.7%)细胞核呈强阳性表达,4例(21.0%)呈中度阳性表达,仅1例(5.3%)呈弱阳性表达。16例(84.2%)阳性细胞弥漫分布,3例(15.8%)呈灶性阳性。对于Ki-67,15例(78.9%)阳性细胞呈全层分布,1例(5.3%)阳性细胞出现在鳞状上皮的上2/3。3例(15.8%)阳性细胞出现在鳞状上皮的下2/3。全部病例细胞核呈强阳性表达。17例(89.4%)阳性细胞弥漫分布,2例(10.6%)呈部分阳性。而萎缩性改变组对p16与Ki-67均呈阴性表达。结论p16与Ki-67是标记绝经后妇女HSIL与萎缩性改变非常有价值的一组抗体,为两者的鉴别诊断提供了更为客观的诊断依据。     

p16INK4A和 p15INK4B对人肝癌细胞增殖和凋亡影响的研究

目的　为探讨抑癌基因 p16 INK4 A和 p15 INK4 B对 Rb基因状态不同的人肝癌细胞系增殖和凋亡的影响。方法　在分析细胞系遗传背景鉴定基础上采用脂质体将构建的 p XJ- p16、p XJ- p15重组质粒转染人肝癌细胞系 BEL74 0 2 (p16 / p15 Rb )和 SMMC772 1(p16 / p15 / Rb- )。应用 PCR、RNA斑点印迹、MTT、集落形成、流式细胞仪等技术检测外源性 p16和 p15基因、m RNA表达及其对肝癌细胞增殖、凋亡的影响。结果　经转染的 BEL74 0 2 - p16 ,BEL74 0 2 - p15细胞 ,分别存在外源性 p16、p15基因 ,在含外源基因细胞中相应的 m RNA表达增强 ;BEL74 0 2 - p15细胞生长速度、集落形成率显著低于对照细胞 BEL 74 0 2 (P<0 .0 1) ;细胞周期分析观察到与亲本细胞比较 ,BEL 74 0 2 - p15的 G1期细胞由 37.7%增高到 4 3.6 % ,S期细胞由 2 2 %下降到 13% (P<0 .0 5 ) ,并出现 G1亚峰 (凋亡峰 )。与此相反 ,BEL74 0 2 - p16细胞增殖未见抑制 ,细胞周期分布无明显差异 ,集落形成率也未见减少。此外 ,SMMC772 1- p16细胞增殖也无抑制。结论　外源性 p15 INK4 B具有抑制人肝癌细胞生长 ,诱导细胞凋亡的作用 ,其作用不受内源性p15影响。而 p16 INK4 A对肝癌细胞生长抑制的作用可能依赖于 RB途径的完整性。     

Ki-67 and ProExC are useful immunohistochemical markers in esophageal squamous intraepithelial neoplasia

Esophageal squamous intraepithelial neoplasia has been widely recognized as a precursor lesion for esophageal squamous cell carcinoma. Early detection offers the best prognosis for esophageal squamous cell carcinoma. The differentiation of squamous dysplasia from reactive change and the classification of squamous dysplasia into high-grade or low-grade are sometimes subjective and challenging. In this study, we sought to evaluate multiple biomarkers and to develop clinically useful adjunct tools for difficult esophageal squamous intraepithelial neoplasia cases. Immunohistochemical stains using antibodies against Ki-67, ProExC, p16, and p53 were performed on esophageal biopsy or resection specimens from 25 patients including 35 foci of high-grade dysplasia and 25 foci of low-grade dysplasia, and from 10 control cases containing 52 foci of normal/reactive hyperplasia. In situ hybridization tests for human papillomavirus were performed in 11 cases. The immunostains for all 4 markers were scored as negative, intermediate, and strong according to established criteria. Intermediate and strong Ki-67 and ProExC staining showed similar detecting power and exhibited very high sensitivity and specificity for distinguishing normal/reactive hyperplasia from esophageal squamous intraepithelial neoplasia and normal/reactive hyperplasia from low-grade esophageal squamous intraepithelial neoplasia. Strong Ki-67 staining was exclusively seen in high-grade esophageal squamous intraepithelial neoplasia, which provided additional value in distinguishing high-grade from low-grade esophageal squamous intraepithelial neoplasia. Strong ProExC staining was also seen in most high-grade esophageal squamous intraepithelial neoplasia foci (80%). Although the frequencies of intermediate/strong staining patterns of p53 increased with increasing degree of dysplasia, the sensitivity of p53 was much lower than that of Ki-67 and ProExC. p16 did not show consistent immunostain pattern in the normal/reactive hyperplasia and esophageal squamous intraepithelial neoplasia. Two (18%) of 11 tested cases were positive for human papillomavirus infection. This study demonstrates that both Ki-67 and ProExC can be used as an adjunct tool for diagnosing difficult cases of esophageal squamous intraepithelial neoplasia.     

子宫颈病变中HPV L1蛋白和p16的表达

目的 研究HPV L1蛋白和p16在子宫颈各种病变中的表达情况,探讨它们在子宫颈病变进展中的预测价值.方法 应用免疫组化方法检测41例各种子宫颈病变(CIN1级18例、CIN2级9例、CIN3级8例和浸润性鳞状细胞癌6例)中HPV L1蛋白和p16的表达.结果 HPV L1蛋白在各种子宫颈病变中的阳性率为26.8%.其中HPV L1在CIN1中的阳性表达率为38.9%,CIN2为44.4%,CIN3和浸润性鳞状细胞癌均无表达.p16在各种子宫颈病变中的阳性率为68.3%,其在CIN1中的阳性表达率为38.9%,CIN2为77.8%,CIN3和浸润性鳞状细胞癌均表达阳性.100%CIN3和浸润性鳞状细胞癌为p16+/HPV L1-,而61.1% CIN1中为p16-/HPV L1+或p16-/HPV L1-.结论 随着子宫颈病变的进展,HPV L1阳性表达率降低而p16阳性表达率增高.p16+/HPV L1-提示子宫颈鳞状上皮内瘤变有进展的可能,而p16-/HPV L1+和p16-/HPV L1-可能为无进展的或潜在消退的子宫颈病变.     

p16INK4a expression as a potential marker of low‐grade cervical intraepithelial neoplasia progression

To evaluate p16^INK4a immunoexpression in CIN1 lesions looking for differences between cases that progress to CIN2/3 maintain CIN1 diagnosis, or spontaneously regress. Seventy‐four CIN1 biopsies were studied. In the follow‐up, a second biopsy was performed and 28.7% showed no lesion (regression), 37.9% maintained CIN1, and 33.4% progressed to CIN2/3. Immunostaining for p16^INK4a was performed in the first biopsy and it was considered positive when there was strong and diffuse staining of the basal and parabasal layers. Pearson's chi‐square was used to compare the groups (p ≤ 0.05). The age of the patients was similar. There was no significant difference in p16^INK4a immunoexpression in the groups, however, statistical analyses showed a significant association when only the progression and regression groups were compared (p = 0.042). Considering p16^INK4a positivity and the progression to CIN2/3, the sensitivity, specificity, positive, and negative predictive values in our cohort were 45%, 75%, 47%, and 94%, respectively. We emphasize that CIN1 with p16^INK4a staining was associated with lesion progression, but the sensitivity was not high. However, the negative predictive value was more reliable (94%) and p16^INK4a may represent a useful biomarker that can identify CIN1 lesions that need particular attention, complementing morphology.     

Relationship of P16 and Ki67 in recurrence of HPV infection and cervical intraepithelial neoplasia

This study aimed to investigate the association of P16 and Ki67 expression in cervical conization with postoperative HPV reinfection and cervical intraepithelial neoplasia. This study retrospectively enrolled patients from January 2012 to December 2013. Patients with negative margins were followed up for 2 years to evaluate the correlation between Ki67 and p16 expression levels in the conization of patients with HPV persistence encountering infection or re-infection and CIN recurrence. The positive expression of p16 and Ki67 was significantly correlated with disease progression (P<0.05). p16 and Ki67 expression was chosen, and results showed that positive expression of p16 and ki67 proteins was a risk factor of disease progression (OR=5.3, 95% CI 1.177~24.365, P=0.042; OR=5.1, 95% CI 1.162~22.387, P=0.031, respectively). Results indicated that routine staining for p16 and Ki67 has clinically significant meaning in guiding disease progress and prognosis at follow-up.     

胃癌中p16INK4a基因启动子高甲基化及其蛋白表达

目的探讨胃癌中p16^INK4a基因失活的主要分子机制及其与胃癌发生、发展的关系。方法采用甲基化特异性PCR技术检测62例胃癌和癌旁组织及10例慢性胃炎黏膜p16^INK4a基因启动子区CpG岛甲基化状态,用免疫组化EnVision两步法检测p16^INK4a蛋白的表达。结果62例胃癌和癌旁组织p16^INK4a基因启动子高甲基化率分别为51．6％（32／62）和19.4％（12／62）,10例正常对照胃黏膜未发现高甲基化,胃癌组织p16^INK4a基因启动子高甲基化率高于癌旁组织和正常对照（P〈0．05）。58.1％（36／62）的胃癌组织p16^INK4a蛋白表达阴性,其中72.2％（26／36）的病例具有p16^INK4a基因启动子的高甲基化,p16^INK4a基因启动子的高甲基化与其蛋白失表达密切相关（P〈0.05）。结论胃癌中p16^INK4a基因启动子的高甲基化是p16^INK4a基因失活的主要机制,并可能是胃癌发生中的早期分子事件。     

The utility of Ki-67 expression in the differential diagnosis of prostatic intraepithelial neoplasia and ductal adenocarcinoma

Cribriform and/or papillary prostatic lesions observed on limited tissue, such as needle biopsy, can pose diagnostic dilemmas. One such area of difficulty is the distinction between papillary and/or cribriform prostatic high-grade prostatic intraepithelial neoplasia (HG-PIN) and ductal adenocarcinoma. Over 48 months, we identified 17 cases of ductal adenocarcinoma and 17 cases of HG-PIN from radical retropubic prostatectomy specimens. The HG-PIN lesions were in all cases associated with an acinar prostatic adenocarcinoma component. For each case, we evaluated the proliferative activity, assessed by Ki-67 immunohistochemistry. The majority (82%) of ductal adenocarcinomas were composed of mixed papillary and cribriform patterns, with the remaining demonstrating pure papillary or cribriform patterns. The HG-PIN lesions showed a papillary, cribriform, or mixed papillary/cribriform architecture. The proliferative activity, defined as Ki-67 labeling index, was statistically higher in ductal adenocarcinoma (mean 33%, range 21%-66%) as compared with HG-PIN (mean 6%, range 2%-15%), with no overlap in the Ki-67 indices (P = 0001). A combination of histological features and measurements of cellular proliferation may be helpful to distinguish HG-PIN from ductal adenocarcinoma in limited prostatic tissue samples.     

p16INK4A and Ki-67 immunostaining on cell blocks from residual ThinPrep material is helpful in identifying significant preneoplastic cervical lesions

Cell blocks can be prepared from residual ThinPrep material, and immunohistochemical staining can be used. The objectives of the current study were (1) to investigate the role of cell block preparation in identifying significant preneoplastic cervical lesions; and (2) to assess the diagnostic value of p16^INK4A and Ki-67 immunostaining on cell blocks to identify significant preneoplastic cervical lesions.Samples from residual substances of ThinPrep from 79 patients were collected for cell block preparations. Cell block sections and histological sections of the same patients were stained with hematoxylin and eosin, and were immunostained with antibodies against p16^INK4A and Ki-67 antigen.The sensitivity and specificity for p16^INK4A and Ki-67 immunostaining to detect high-grade squamous intraepithelial neoplasia were 95.12% and 73.68%, respectively. The positive predictive value and negative predictive value were 79.59% and 93.33%, respectively. Immunostaining of cell blocks for p16^INK4A and Ki-67 exhibited a statistically significant association with the presence of significant lesions on cell blocks (P < 0.05).p16^INK4A and Ki-67 immunostaining on cell blocks from residual ThinPrep material is helpful in identifying significant preneoplastic cervical lesions.