胃癌组织中Tenascin、MVD、Ki-67的表达及其临床意义

目的：探讨胃癌组织中Tenascin蛋白、微血管密度（microvascular density,MVD）、Ki-67的表达及其与临床病理特征的关系。方法：采用免疫组化Elivision法检测70例胃癌组织和20例癌旁正常组织中Tenascin、CD34和Ki-67的表达。结果：①正常胃黏膜上皮Tenascin阴性,胃癌中的Tenascin主要表达于肿瘤相关纤维母细胞的胞质中,且与胃癌的Lauren分型、分化程度、临床分期、淋巴结转移显著相关（P〈0．05）;②胃癌中MVD和Ki-67-LI（标记指数）均高于正常胃黏膜（P〈0．001）,且均与胃癌的临床分期、浸润深度、淋巴结转移显著相关（P〈0．05）;结论：胃癌组织中Tenascin可抑制胃癌的演进,MVD及Ki-67可作为胃癌患者预后的预测指标,联合检测胃癌组织Tenascin、MVD、Ki-67的表达情况,对于进一步了解胃癌的生物学行为和判断预后具有一定的临床价值与意义。     

PAX-5、CD117、PDGFRA在胃肠道神经内分泌肿瘤中的表达及意义

目的研究PAX-5、CD117、PDGFRA在胃肠道神经内分泌肿瘤中的表达,并分析与肿瘤生物学特点及患者生存时间的关系。方法应用组织芯片及免疫组化法,对96例胃肠道神经内分泌肿瘤的免疫表型和预后进行分析。结果 PAX-5、CD117、PDGFRA蛋白的阳性率分别为31.3%、19.8%、53.1%。PAX-5和PDGFRA表达与胃肠道神经内分泌肿瘤类型、肿瘤大小、TNM分期及淋巴结转移有显著相关性,CD117表达与TNM分期及淋巴结转移有显著相关性。TNM分期、淋巴结转移、肿瘤大小、CD117、P53、Ki-67表达与生存时间呈负相关,但只有TNM分期、CD117表达和Ki-67高表达为影响预后的独立因素。结论 PAX-5、CD117、PDGFRA的表达可能与胃肠道神经内分泌肿瘤的发生发展有关,CD117表达和Ki-67高表达是独立预后因子,PAX-5可以作为潜在的分子治疗靶点。     

CD117、CD34、DOG-1、WT-1 在胃肠道间质瘤中的表达及意义

目的:探讨DOG-1、CD117、CD34、WT-1在胃肠道间质瘤(GIST)中的表达及临床意义。方法:应用免疫组织化学SP法检测DOG-1、CD117、CD34、WT-1在39例GIST患者肿瘤组织中的表达。结果:GIST光镜下主要由梭形细胞和(或)上皮样细胞或多形性细胞混合或单一性组成。DOG-1、CD117、CD34和WT-1在GIST肿瘤组织中阳性表达率分别为92.3%(36/39)、71.7%(28/39)、64.1%(25/39)、23.1%(9/39),四者的阳性表达率在各风险程度组(极低及低度危险性GIST、中度危险性GIST、高度危险性GIST)两两比较中差异均无统计学意义(P>0.05)。DOG-1与CD117相比,在极低及低度危险组中表达有显著差异(P<0.05),DOG-1与CD34相比,在极低及低度危险组、高度危险组中表达有显著差异(P<0.05);WT-1与CD117相比,在中度及高度危险组中表达有显著差异(P<0.05),WT-1与CD34相比,在中度危险组中表达有显著差异(P<0.05)。对照组平滑肌瘤、纤维瘤、神经鞘瘤中CD117、DOG-1在GIST中的表达明显高于其他多种梭形细胞的表达(P<0.05)。结论:DOG1是GIST较为敏感和特异的标记物,WT-1在极低及低度危险性GIST中有一定的诊断价值,在GIST的诊断中加入两者将使患者更加受益。     

胃癌组织中Tenascin、MVD、Ki-67 的表达及其临床意义

目的:探讨胃癌组织中Tenascin蛋白、微血管密度(microvascular density,MVD)、Ki-67的表达及其与临床病理特征的关系。方法:采用免疫组化Elivision法检测70例胃癌组织和20例癌旁正常组织中Tenascin、CD34和Ki-67的表达。结果:①正常胃黏膜上皮Tenascin阴性,胃癌中的Tenascin主要表达于肿瘤相关纤维母细胞的胞质中,且与胃癌的Lauren分型、分化程度、临床分期、淋巴结转移显著相关(P<0.05);②胃癌中MVD和Ki-67-L(I标记指数)均高于正常胃黏膜(P<0.001),且均与胃癌的临床分期、浸润深度、淋巴结转移显著相关(P<0.05);结论:胃癌组织中Tenascin可抑制胃癌的演进,MVD及Ki-67可作为胃癌患者预后的预测指标,联合检测胃癌组织Tenascin、MVD、Ki-67的表达情况,对于进一步了解胃癌的生物学行为和判断预后具有一定的临床价值与意义。     

神经胶质瘤中PTEN、Ki-67和HIF-1α的表达及临床意义

目的:抑癌基因PTEN、癌基因Ki-67及HIF-1α对多种人类肿瘤的恶性进展均起重要的调控作用。本研究主要探讨PTEN、Ki-67及HIF-1α在人脑胶质瘤中的表达及临床意义,为胶质瘤患者预后的判定、分子病理学的诊断、基因靶向的治疗奠定理论基础。方法:在83例原发性人脑胶质瘤组织样本中,通过免疫组化的方法检测PTEN、Ki-67及HIF-1α的表达情况,并分析其表达相互间及其表达与肿瘤恶性级别之间的相关性。结果:在正常脑组织中,PTEN的表达均为阳性,Ki-67的表达均为阴性,10%(1/10)的样本HIF-1α的表达为阳性。在胶质细胞瘤中,PTEN的表达显著降低(P=0.001),而Ki-67(P0.001)和HIF-1α(P=0.001)的表达明显增高。随肿瘤恶性级别的增高,PTEN的表达呈降低趋势(P0.001),而Ki-67和HIF-1α的表达呈升高趋势(两者P均0.001)。相关性分析表明,PTEN的表达与Ki-67和HIF-1α的表达呈负相关(r值分别为-0.289和-0.304;P值分别为0.008和0.005),Ki-67的表达与HIF-1α的表达呈正相关(r=0.833;P0.001)。结论:胶质瘤组织缺乏抑癌基因PTEN蛋白的表达,而高度表达癌基因Ki-67和HIF-1α。抑癌基因PTEN表达减少或失活,癌基因Ki-67和HIF-1α的过表达对胶质瘤恶性进展可能起到至关重要的作用。PTEN、Ki-67和HIF-1α蛋白的联合检测对胶质瘤恶性程度和预后的判定有十分重要的临床意义。     

侵袭性前列腺癌的CT、MRI表现与生物因子表达及淋巴结转移的相关性研究

摘要 目的：探讨侵袭性前列腺癌的电子计算机断层扫描（CT）、磁共振成像（MRI）表现与生物因子表达及淋巴结转移的相关性研究。方法：收集2016年2月-2021年2月于我院确诊前列腺癌患者117例,所以患者均行前列腺根治切除+盆腔淋巴结清扫术,术前均完善CT、MRI影像学检查,根据术后病理结果、Gleason评分分为侵袭性前列腺癌组（63例）和非侵袭性前列腺癌组（54例）,分析患者CT、MRI表现特征。采用免疫组织化学染色分析前列腺特异抗原（PSA）、增殖细胞核抗原（PCNA）、血管内皮生长因子（VEGF）、细胞核增殖相关抗原 (Ki-67)及肿瘤抑制蛋白（P53）的阳性表达情况。分析侵袭性前列腺癌的CT、MRI表现与生物因子表达及淋巴结转移之间的关系。结果：侵袭性前列腺癌患者CT、MRI均可见不同程度的前列腺异常改变,主要表现为结节状、波浪状突起,边缘模糊、毛糙,前列腺癌突破包膜并累及邻近精囊腺、膀胱、直肠及周围脂肪组织,可见盆腔肿大淋巴结转移,测量表观扩散系数（ADC）值及动态增强MRI定量参数可大致区分病灶良恶性及恶性程度分级情况。侵袭性前列腺癌组PSA的表达低于非侵袭性前列腺癌组,而PCNA、VEGF、Ki-67、P53的表达及淋巴结转移高于非侵袭性前列腺癌组,差异均有统计学意义（P＜0.05）。侵袭性前列腺癌患者肿瘤边缘表现为模糊、清晰时PSA、PCNA阳性表达率及淋巴结转移率组间差异均有统计学意义（P＜0.05）;肿瘤直径≤2 cm、＞2 cm时的淋巴结转移率组间差异具有统计学意义（P＜0.05）。侵袭性前列腺癌组ADC值低于非侵袭性前列腺癌组,而Kep和Ktrans高于非侵袭性前列腺癌组患者,差异均有统计学意义（P＜0.05）。侵袭性前列腺癌患者ADC值与PSA阳性表达呈正相关性（P＜0.05）,与PCNA、Ki-67、P53阳性表达及淋巴结转移呈负相关（P＜0.05）。侵袭性前列腺癌患者Kep、Ktrans与PSA阳性表达呈负相关性（P＜0.05）,与PCNA、Ki-67、P53阳性表达及淋巴结转移呈正相关（P＜0.05）。结论：侵袭性前列腺癌CT、MRI形态学具有一定特征性,通过测量ADC值及动态增强MRI定量参数可大致区分病灶恶性程度情况,初步评估前列腺癌的生物学行为,为指导临床综合治疗及预后情况提供客观依据。     

三阴乳腺癌新辅助化疗的预后与Ki-67、TOPK的表达情况相关性研究

摘要 目的：研究三阴乳腺癌（TNBC）新辅助化疗（NACT）的预后与Ki-67、TOPK的表达情况的相关性。方法：回顾性收集2014年3月至2016年6月期间我院收治并行NACT的TNBC患者90例作为研究对象,免疫组化法检测肿瘤组织中Ki-67和TOPK的表达,分析两者的表达与患者临床资料的关系,应用COX回归模型进行生存分析,分析影响TNBC患者预后的独立危险因素。结果：在治疗情况方面,91.11 %（82/90）的患者接受了≥4个周期的NACT治疗,84.44 %（76/90）的患者接受了手术治疗,术后pCR为22.22 %（20/90）,ORR为75.56 %（68/90）。为期5年的随访中,90例患者中共死亡8例（死于TNBC者6例,其他原因2例）,失访2例;治疗后Ki-67和TOPK的阳性率均显著低于治疗前（P＜0.05）;Ki-67、TOPK阳性和阴性表达病例在不同TNM分期、淋巴结是否转移上具有显著性差异（P＜0.05）;Ki-67和TOPK阳性表达患者的无进展生存期（PFS）和总生存期（OS）显著短于阴性表达者（P＜0.05）;TNM分期、淋巴结转移、Ki-67和TOPK阳性表达均为影响TNBC患者预后的独立危险因素。结论：Ki-67和TOPK在TNBC组织中的表达与患者的病理学特征及预后密切相关,是影响预后的独立危险因素。     

抗凋亡蛋白Survivin和Ki-67在肝癌组织中的表达及相关性研究

目的:观察抗凋亡蛋白Survivin和Ki-67在原发性肝癌(PHC)中的表达并探讨其临床意义.方法:采用免疫组织化学S-P法检测34例原发性肝癌组织、14例癌旁正常肝组织中survivin和Ki-67的表达情况.结果:原发性肝癌组织中survivin的阳性表达定位于细胞浆和细胞核,阳性表达率明显高于癌旁正常肝组织(P＜0.05);Ki-67的阳性表达主要定位于细胞核,其阳性表达率亦明显高于癌旁正常肝组织(P＜0.05).二者的阳性表达与原发性肝癌患者的性别、年龄、肿瘤大小无相关性(P＞0.05),与淋巴结转移、组织分化程度及癌栓是否形成有关(P＜0.05);此外,二者在原发性肝癌组织中的表达呈显著正相关(P＜0.05).结论:Survivin和Ki-67在原发性肝癌组织中表达上调,均与原发性肝癌的组织分化程度及癌栓是否形成密切相关;检测survivin和Ki-67的表达有助于原发性肝癌的预防、治疗和预后评估.     

胃肠道间质瘤组织S100A4、CD55的表达及与临床病理参数及预后的关系研究

摘要 目的：探讨胃肠道间质瘤组织S100A4、衰变加速因子（CD55）的表达及与临床病理参数及预后的关系。方法：选取2011年1月-2015年12月在陆军军医大学附属西南医院江北院区/陆军第958医院行手术切除并经术后病理确诊的124例GIST组织和对应的瘤旁正常组织病理蜡块,采用免疫组化法检测S100A4、CD55在GIST组织及瘤旁正常组织中的表达,分析其表达与GIST患者临床病理参数及预后的关系。结果：S100A4、CD55在GIST组织中的高表达率明显高于瘤旁正常组织,差异有统计学意义（P<0.05）。S100A4、CD55表达与GIST患者肿瘤大小、核分裂相、NIH分级及远处转移有关（P<0.05）,与GIST患者年龄、性别、发生部位、组织学类型无关（P>0.05）。S100A4、CD55低表达的GIST患者术后复发率和转移率低于S100A4、CD55高表达的GIST患者,差异有统计学意义（P<0.05）;S100A4、CD55低表达组患者5年中位无进展生存期明显高于S100A4、CD55高表达组,差异有统计学意义（P<0.05）。结论：S100A4、CD55在GIST组织存在高表达, 二者可能参与了GIST的侵袭与转移,并与患者术后的预后有关。     

能谱CT成像对甲状腺癌局部浸润深度的诊断价值及其定量参数与肿瘤组织中Ki67、VEGF、CD34、EGFR的相关性

摘要 目的：探讨能谱CT成像对甲状腺癌局部浸润深度的诊断价值及其定量参数与肿瘤组织中Ki67、VEGF、CD34、EGFR的相关性。方法：回顾性分析2018年6月-2021年6月我院经手术或穿刺活检病理证实为甲状腺肿瘤性病变的患者96例，其中29例为甲状腺癌局部浸润组（A组），34例为甲状腺癌无浸润组（B组），33例为甲状腺腺瘤组（C组）。另取56例甲状腺另一侧叶正常组织作为对照组（D组）。所有患者均完善能谱CT检查，采集图像后在能谱CT Viewer分析软件上测量病变区碘浓度，计算能谱曲线斜率。采用免疫组织化学染色分析Ki-67、VEGF、CD34、EGFR的表达情况。采用Spearman秩相关分析评价碘浓度、能谱曲线斜率与甲状腺癌肿瘤组织中Ki-67、VEGF、CD34、EGFR表达的相关性。结果：在平扫、动脉期、静脉期，A组、B组、C组和D组的碘浓度逐渐增大，两两比较差异均有统计学意义（P＜0.05）。甲状腺癌局部浸润组织能谱曲线呈"低平型"，斜率为较小负值，正常甲状腺组织能谱曲线为下降型，斜率为负值；在平扫、动脉期、静脉期，A组、B组、C组和D组的能谱曲线斜率逐渐变小，差异均有统计学意义（P＜0.05）。A组中Ki-67、VEGF、CD34和EGFR的阳性表达率均高于B组和C组，差异均有统计学意义（P＜0.05）。碘浓度在动脉期、静脉期与Ki-67、VEGF、CD34、EGFR表达呈正相关（P＜0.05），碘浓度在平扫与Ki-67表达呈正相关（P＜0.05），碘浓度在平扫与VEGF、CD34、EGFR表达无相关性（P＞0.05）。能谱曲线斜率在动脉期、静脉期与Ki-67、VEGF、CD34、EGFR表达呈正相关（P＜0.05），能谱曲线斜率在平扫与VEGF表达呈正相关（P＜0.05），能谱曲线斜率在平扫与Ki-67、CD34、EGFR表达无相关性（P＞0.05）。结论：能谱CT成像检查对甲状腺癌局部浸润深度的判断具有重要的价值，其定量参数碘浓度、能谱曲线斜率与Ki67、VEGF、CD34、EGFR具有相关性，可间接反映肿瘤微血管、肿瘤血管生成、甲状腺癌分化程度、浸润程度等情况，对评价甲状腺癌生物学行为可提供有价值的信息。     

CD133在胃肠间质瘤的表达及其临床意义

目的 探讨胃肠间质瘤（gastrointestinal stromal tumor,GIST）中的CD133的表达及其与GIST临床病理特征的关系.方法 采用免疫组织化学法,检测122例胃肠间质瘤患者组织中的CD133、CD117、CD34、DOG-1、KI-67的表达情况.结果 CD133、CD117、CD34 、DOG-1、KI-67的阳性表达率分别为74.6%（91/122）、98.4%（120/122）、86.9%（106/122）、95.1%（116/122）、47.5%（58/122）.CD133的表达水平与胃肠间质瘤危险度分组高低、核分裂像数目、肿瘤部位有关（P〈0.05）,而与患者性别、年龄、肿瘤大小无明显相关性（P〉0.05）.CD133的表达水平与DOG-1的表达水平无明显相关性（P〉0.05）,而与CD117、CD34、KI-67的表达水平呈正相关.结论 CD133蛋白的表达可能与GIST的恶性行为与预后有关,与CD117、CD34和DOG-1联合检测对于判断GIST的病理性质可能具有重要价值.     

Prediction of aggressiveness of gastrointestinal stromal tumours based on immunostaining with bcl-2, Ki-67 and p53

OBJECTIVE: While the use of fine needle aspiration (FNA) in the diagnosis of gastrointestinal stromal tumours (GISTs) is well-established, it can be difficult to predict the prognosis of GIST based on morphology alone. The objective of the current study was to determine if expression of bcl-2, Ki-67 and p53 correlated with the outcome of GISTs based on cytological material. METHODS: Cell-blocks from 14 GISTs diagnosed by FNA were retrieved. Immunostaining was performed with antibodies against bcl-2, Ki-67 and p53. All cytological diagnoses were confirmed by positive immunostaining with c-kit and/or subsequent histological evaluation. Positivity for bcl-2, Ki-67 and p53 was defined as the presence of > or =10% cytoplasmic staining, > or =5% nuclear staining and > or =5% nuclear staining respectively. RESULTS: The 14 patients consisted of seven males and seven females with a mean age of 58 years. The average follow-up interval was 46 months. Six had a benign course and eight developed recurrences/metastases. Thirteen (93%) cases showed positive staining for bcl-2. Positive Ki-67 and p53 staining was noted in one (7%) and seven (50%) cases respectively. The difference in staining for p53 between aggressive and non-aggressive GISTs was statistically significant. No statistically significant difference was noted for bcl-2 staining or Ki-67 labelling index between the two groups. CONCLUSIONS: According to our observations, p53 immunostaining may be useful in predicting the outcome of GIST diagnosed by FNA; Ki-67 and bcl-2 are not useful as prognostic markers for GIST in FNA specimens.     

PCNA Ki-67 dissociation in childhood acute lymphoblastic leukaemia. An Immunofluorescent Laser Confocal Scanning Microscopical study.

Cell proliferation rates of diagnostic marrow aspirate cells of 21 children with Acute Lymphoblastic Leukaemia and 16 controls were compared using immunocytochemical labelling of PCNA and Ki-67 antigen as assessed by Confocal Laser Scanning Microscopy. The results showed an unexpected, highly significant degree of dissociation between PCNA and Ki-67 expression in ALL blasts. The PCNA labelling indices of ALL patients were significantly increased (mean 44, range 24-77) compared with normal reactive marrow cells (mean 13.8, range 4-26) (p<0.000001, Mann Whitney U two tailed test), suggesting an abnormal commitment to proliferation. Ki-67 expression was raised to a lesser extent in ALL cells (mean 14.8, range 1.2-35) when compared to non-malignant proliferations (mean 6.6, range 1.7-25) (p < 0.02). PCNA/Ki-67 LI ratios in ALL (mean 7, range 1.1-35) were higher than in controls (mean 2.7, range 1.04-6.5, p<0.09). As cell proliferation rates actually achieved in the bone marrow do not differ as strongly as suggested by the extreme difference in PCNA labelling, a pathological dissociation of PCNA / Ki-67 expression exists, suggesting immortalisation.     

Mitosin,a novel marker of cell proliferation and early recurrence in intracranial meningiomas

The expression of mitosin, a novel proliferation-associated molecule was evaluated immunohistochemically in a consecutive series of 47 patients with primary intracranial benign and atypical meningiomas. Mitosin expression was correlated with proliferation markers Ki-67 (MIB-1), proliferating cell nuclear antigen (PCNA), topoisomerase IIalpha (TopoIIalpha) and mitotic index, as well as with standard clinicopathological parameters and patient outcome. Seven tumors recurred (14.8%) following gross total resection, within a follow-up period ranging from 21 to 108 months (median 60 months). The higher proliferation indices were obtained with mitosin and PCNA and the lower ones with TopoIIalpha. Mitosin labeling index (LI) ranged from 0.1 to 57% (median 3%), with a significant overlapping of values between grades. A significant positive correlation was shown between mitosin LI on the one hand and Ki-67 LI (p < 0.001), or the mitotic index (p = 0.027) on the other. The incidence of recurrence was higher in cases with a mitosin LI higher than 3% (p = 0.048). Univariate analysis disclosed mitosin LI (p = 0.033) along with the mitotic index (p = 0.024) and tumor size (p = 0.028) as significant predictors of shortened recurrence-free survival. In multivariate analysis, the labeling indices of mitosin (p = 0.035) and Ki-67 (p = 0.032), along with tumor size, were shown to provide independent prognostic information, beyond that obtained by standard clinical and pathological parameters. However, as indicated by factor analysis, the prognostic information yielded by mitosin was superior to that provided by the remaining proliferation markers (p = 0.041). We conclude that mitosin immunohistochemical expression, although failing to discriminate between benign and atypical meningiomas, may be of use as a novel cell proliferation marker and as a predictor of tumor recurrence.     

Immunohistochemical evaluation of bcl-2, ER-alpha,caspase -3, -8, -9, PCNA and Ki-67 expressions in canine mammary carcinomas

We investigated the expression of bcl-2, estrogen receptor alpha (ER-alpha), caspase-3, ?8, ?9, proliferating cell nuclear antigen (PCNA) and Ki-67 in canine mammary carcinomas. We used 65 paraffin embedded and re-diagnosed archival canine mammary tumor samples to which we applied the routine streptavidin-biotin-peroxidase technique. Seventeen cases were re-diagnosed as tubulopapillary carcinoma, 31 were re-diagnosed as complex carcinoma and 17 were re-diagnosed as carcinosarcoma. Differences of expression of bcl-2 and PCNA were statistically significant according to tumor type. Differences in expression of ER-alpha, caspase-3, ?8, ?9 and Ki-67 were not statistically significant. Differences of expression of bcl-2 and PCNA were statistically significant compared to ER-alpha, caspase-3, ?8, ?9 and Ki-67 in carcinosarcomas. We report the prognostic significance of bcl-2 and PCNA expression in canine mammary carcinosarcomas.     

人乳头状瘤病毒感染与宫颈癌患者临床病理特征和Ki-67、PCNA的关系

目的:研究人乳头状瘤病毒(HPV)感染与宫颈癌患者临床病理特征和Ki-67、细胞增殖抗原(PCNA)的相关性,从而为临床宫颈癌的诊治提供参考依据。方法:选取2016年3月～2018年6月于我院接受手术治疗的宫颈病变患者130例为研究对象。其中宫颈癌患者30例记为宫颈癌组,宫颈上皮内瘤变患者68例记为宫颈上皮内瘤变组,慢性宫颈炎患者32例记为对照组。采用免疫组织化学法检测各组宫颈组织中HPV感染、Ki-67以及PCNA阳性表达情况,并分析HPV与宫颈癌患者临床病理特征的关系及其与Ki-67、PCNA的相关性。结果:宫颈癌组、宫颈上皮内瘤变组患者HPV、Ki-67以及PCNA阳性率均高于对照组,宫颈癌组高于宫颈上皮内瘤变组(均P0.05)。临床分期Ⅲ～Ⅳ期以及淋巴结转移宫颈癌患者HPV感染率均明显高于临床分期Ⅰ～Ⅱ期与无淋巴结转移患者(均P0.05)。经Spearman相关性分析可得:宫颈癌患者HPV感染与Ki-67、PCNA表达均呈正相关关系(均P0.05)。结论:宫颈癌患者存在明显的HPV感染,且HPV感染与宫颈癌患者临床分期、淋巴结转移、Ki-67、PCNA表达存在一定相关性,临床可通过对HPV、Ki-67、PCNA进行联合检测,从而有助于宫颈癌的早期诊断。     

Image analysis of in situ cell cycle related changes of PCNA and Ki-67 proliferating antigen expression

Abstract. This study reports on the proliferating cell nuclear antigen (PCNA) and Ki-67 cell cycle related expression and distribution pattern analysed in the same cells. MCF-7 cells were synchronized by mitotic detachment and triple stained for DNA, PCNA and Ki-67. The major cell type was identified on each time sample as a function of the PCNA/Ki-67 pattern, and both antigens as well as DNA were quantified. During G₁ phase, the expression of PCNA greatly increased whereas Ki-67 content decreased. During S phase, nuclear Ki-67 content continuously increased especially in the second half of this phase, mainly due to the accumulation of the antigen in the nucleoli. During G₂ phase, the antigen significantly passed into the nucleoplasm, its content continued to increase and reached its maximum in mitotic cells. Nuclear PCNA content mostly increased in the first part of S phase and sharply declined in mitotic cells as the antigen shifted to the cytoplasm. Cells showing PCNA positive Ki-67 negative labelling were observed in all time samples from the beginning of the experiment. Their nuclear size, DNA content (of G₁ cells), PCNA content (equivalent to the content of some late G, cells) and time occurrence (their percentage increased after the last late G₁ cells had disappeared) tend to indicate that these cells have left the cycle by the end of G₁ phase to enter a quiescent state. Cells coming out of mitosis split into two groups according to their Ki-67/PCNA content. The biggest fraction was PCNA negative and Ki-67 positive while the smallest showed positive staining for both antibodies. Cells of this second cohort slowly lost their 1–67 while their PCNA content increased as they moved through G₁. Concurrently, most of the cells of the first cohort (here called Q2 and Q3 cell types) lost their Ki-67 without increasing their PCNA content; then they joined cells of the second cohort by increasing their PCNA content at the end of G, phase. Some cells of this first cohort can also increase their PCNA and thus reach cells of the first cohort before the end of G₁ phase. The existence of these two main cell cohorts suggests that cells after mitosis differ in some way that make them progress dlfferently through G₁. Some cells seem to go through early G₁ (G_1a and late G₁ (G_lb) while others may come out of mitosis committed to go through the following cycle by directly entering late G₁ compartment.     

Reclassification of gastrointestinal mesenchymal tumors

A retrospective (1985-2005) multicentric evaluation of the pathological diagnoses of GIST (gastrointestinal stromal tumor) in Hungary was performed. A large cohort of 463 patients with abdominal mesenchymal tumors was reevaluated with the help of immunohistochemistry for CD117, CD34, desmin and S100. Prognostic groups have been established based on international criteria. Two hundred fifty five GISTs have been found in 245 patients during the evaluated period. Beside GIST, 81 leiomyogenic and 25 neurogenic tumors were the primary pathological diagnosis and in 74/255 cases the primary diagnosis was a benign tumor. In our cohort high-risk GIST cases were found to be the predominant. Based on our experience, in case of abdominal tumors of uncertain localization, immunohistochemical markers of CD117, CD34, S100 and desmin provide a proper tool for precise identification of the tumor entity including GIST.     

Simultaneous flow cytometric analysis of surface markers and nuclear Ki-67 antigen in leukemia and lymphoma

The monoclonal antibody Ki-67 identifies an antigen present during the late G1, S, G2, and M phases of the cell cycle, whereas resting cells do not express this antigen. Immunostaining with Ki-67 provides a simple method with which to determine the growth fraction of a malignant cell population without requiring a laborious procedure or use of radioactive materials. Thus far, detection of Ki-67-positive cells by flow cytometry was limited because of nuclear location of the antigen. In this study, periodate-lysine-paraformaldehyde (PLP) fixation of cells in suspension, labeling with Ki-67, and the subsequent flow cytometric analysis of the tumor growth fraction is described. Fixation with PLP at -10 degrees C for 15 min rendered the plasma membrane permeable without destroying cell surface antigens. Thus double immunofluorescence studies using both a surface marker and Ki-67 could be performed. This offers the additional advantage of being able to define the phenotype of proliferating cells. This method was applied to determine the growth fraction in peripheral blood and bone marrow samples of patients with leukemia and non-Hodgkin's lymphoma. The results of Ki-67 studies in 91 patients are shown. A wide variability of individual Ki-67 values was observed within each entity. Use of this flow cytometric procedure substantially facilitates the quantification of proliferating cells in pathological blood and bone marrow samples.