Overexpression of p53 associated with tumor angiogenesis, tumor cell proliferation, and prognosis in gastric carcinoma

Mutations in the p53 gene seem to be the most common genetic change in human malignancies. Recently, it was reported that p53 mutation was significantly associated with prognosis in various cancers. In this study, we investigated the correlation between p53 overexpression and prognosis of gastric carcinoma using immunohistochemical staining with an anti-p53 antibody. Although there was no significant association between p53 status and various clinicopathologic factors, prognosis of patients with p53-positive tumors was significantly worse than of those with p53-negative: tumors. Both microvessel count (MVC; the mean number of microvessels in the five areas of highest vascular density at 200x magnification) and PCNA labeling index (PCNA LI; percentage of positive cells per more than 500 tumor cells) were significantly higher in p53-positive tumors than in p53-negative tumors. In summary, it is suggested that p53 overexpression is closely associated with tumor angiogenesis, tumor cell proliferation and prognosis of gastric carcinoma.     

Microvessel count, proliferating cell nuclear antigen and Ki-67 indices in gastric adenocarcinoma

The aim of the present study was to immunohistochemically investigate the prognostic value of neovascularization (expressed as microvessel count-MVC) and tumor cell proliferation (expressed as PCNA labeling index PLI and Ki-67 labeling index KLI) in gastric adenocarcinoma. Correlations with clinicopathologic features were also evaluated. Tumor specimens from 74 patients diagnosed as gastric adenocarcinoma were included in this study. Formalin fixed, paraffin embedded tissue sections stained immunohistochemically with F-VIII, PC10 and MIB-1 monoclonal antibodies. By ocular grid subdivided into 100 areas, number of microvessels and PC10, MIB-1 positive and negative cells were counted at x400 magnification. Chi-square test, Kaplan-Meier method and cox regression analysis were used for statistical analysis. The results showed that, MVC and PLI had a significant correlation with invasion and lymph node metastasis. The prognosis was significantly worse in patients with high MVC (>14 ) and with high PLI (>49%). However any relationship was not observed between KLI (38%) and clinicopathologic parameters, so KLI failed to predict the prognosis. Cox model showed that, MVC and PLI were independent prognostic variables. Ki-67 labeling index in gastric carcinomas has no prognostic relevance. However, the evaluation of microvessel count and proliferating cell nuclear antigen index in gastric carcinomas could be reliable indicators of prognosis.     

PCNA及DNA含量与胃癌预后关系的研究

作者利用免疫组化LSAB法和流式细胞分析技术（FCM）对41例进展期胃癌组织中增殖细胞核抗原（PCNA）的表达及细胞核DNA含量进行了测定。结果表明：PCNA标记指数及DNA非整信体检出率与胃癌病人术后5年生存率呈明显负相关（P＜0．05），与淋巴结转移呈正相关（P＜0．05）。DNA非整信体检出率与PCNA标记指数具有良好的相关性，二者均可用于判断胃癌预后，而二者同时运用则更有参考价值。     

Clinicopathological and prognostic significance of thymidine phosphorylase and proliferating cell nuclear antigen in gastric carcinoma

We have reported that thymidine phosphorylase (dThdPase) is identical to platelet derived-endothelial cell growth factor and that it has an angiogenic activity in vitro and in human carcinoma tissues as well as gastric carcinoma. Recently, we revealed that dThdPase may have an another function(s) besides angiogenesis in vitro and in human solid tumors. Using immunohistochemistry, we examined retrospectively whether the expression of dThdPase was correlated with tumor growth, comparing it with the proliferating cell nuclear antigen labeling index (PCNA LI) and examining their prognostic significance in 116 patients with gastric carcinoma. A direct correlation of these two factors was observed (R=0.659, P<0.001). A multivariate Cox regression analysis showed that both dThdPase positivity and PCNA LI were independent prognostic factors, as were depth of invasion and lymph node metastasis. Furthermore, the patients with dThdPase-positive/high PCNA LI tumors had the worst prognoses. The combination of dThdPase and PCNA expression is a better tool for predicting the prognosis of patients with gastric carcinoma than the expression of either of them alone. These results raise the possibility that dThdPase may have a function(s) involved in tumor growth besides angiogenesis.     

Relationship between immunohistochemical evaluation of thymidylate synthase and proliferating cell nuclear antigen labeling index in gastrointestinal carcinoma

We undertook this study to clarify the effect of immunohistochemical expression of thymidylate synthase (TS) on proliferative activity of carcinoma lesions in patients with gastrointestinal carcinoma. TS was immunohistochemically evaluated in 53 gastric carcinoma and 51 colorectal carcinoma patients using anti-TS polyclonal antibody. Proliferative activity represented by proliferating cell nuclear antigen (PCNA) labeling index (LI) was also immunohistochemically estimated using monoclonal antibody PC10. Then, the correlation between TS expression and PCNA LI was investigated. Both in gastric and colorectal carcinoma, the PCNA LIs of the high-TS group were significantly higher than those of the low-TS group. In gastric carcinoma, the PCNA LIs of the high-TS group were higher than those of the low-TS group in differentiated adenocarcinoma, in the depth of mucosal and/or submucosal layer, in cases without lymph node metastasis, and notwithstanding lymphatic or venous invasion. In colorectal carcinoma, PCNA LIs of the high-TS group were higher than those of the low-TS group in well differentiated adenocarcinoma, in the depth of serosa, in cases with lymph node metastasis, in cases with lymphatic invasion, and notwithstanding venous invasion. Immunohistochemical expression of TS was correlated with the proliferative activity represented by PCNA LI, but was not identical with PCNA LI.     

Proliferation kinetics and prognosis in gastric cancer after resection

The influence of proliferation and proliferation kinetics on prognosis in gastric cancer after complete resection are controversial. In a prospective study we investigated the tumour specimens of 111 patients after resection of gastric cancer, who received 200 mg intravenous (i.v.) bromodeoxyuridine (BrdU) pre-operatively. The following biological parameters were analysed in the tumour tissue using flow-cytometry: DNA ploidy, proportion of S-phase cells, BrdU labelling index (LI), DNA synthesis time (T(s)), potential tumour doubling time (T(pot)), proliferating cell nuclear antigen (PCNA) and Ki-67 LI. The median follow-up time was 40 months (range 19-62 months). Besides the established pathohistological prognostic factors, univariate analysis revealed a prognostic influence on survival for BrdU LI, T(pot) and the proportion of S-phase cells. By multivariate Cox analysis of the completely resected cases, only tumour stage and T(pot) had a significant, independent influence on survival. By classification and regression trees (CART) analysis, resection status, tumour stage and T(pot) defined risk groups with significantly different outcomes. A short T(pot) was a predictor of better survival in stage I, II and IIIA tumours. Ploidy and the other investigated proliferation-related parameters failed to demonstrate any influence on prognosis after resection of gastric cancer.     

食管癌微血管定量及PCNA表达的临床意义

[目的］检测食管癌CD34和PCNA的表达,探讨微血管密度和细胞增殖状态与食管癌临床病理的关系。[方法］运用免疫组化法对30例食管癌手术标本进行CD34和PCNA检测。［结果］CD34标记的微血管在食管癌中呈异质性。高分化、早期食管癌微血管密度明显低于低分化、中晚期食管癌（P＜0.05和P＜0.01）,同样PCNA标记指数在高分化和早期食管癌中明显较低（P＜0.05和P＜0.01）。存活5年以上与5年以内复发死亡者之间微血管密度和PCNA指数差异有显著性意义（P＜0．01）。［结论］微血管定量和PCNA对食管癌预后判断有重要意义。     

Proliferating Cell Nuclear Antigen Immunostaining in Breast Cancer and Its Relation to Prognosis

Proliferating cell nuclear antigen (PCNA) appears in the cellnuclei during the late G1 to S phases of the cell cycle andis thought to be closely related to cellular proliferation.The authors have conducted an immunohistochemical study in orderto investigate the tissue expression of PCNA and its clinicalsignificance in breast cancer. Excluding cases with absolutelyno positive cells on the section specimen, the mean value (%)for the PCNA labeling index (LI) was 30.4 in 187 cases of invasiveductal carcinoma. No correlations between PCNA LI and any clinicopathologicalfactors such as tumor diameter and tumor stage were observed.Also, no significant correlation was observed with Ki-67 LI.A positive correlation was, however, observed with the tissueexpression of c-erbB-2 protein. We divided 82 patients withstage II invasive ductal carcinoma into PCNA LI of <10, PCNALI of 10–50 and PCNA of >50, and analyzed the specimensfor any correlation with prognosis. The group with PCNA LI of>50 had significantly poorer prognoses than the other groups.From the above, we concluded immunostaining for PCNA to be usefulas a prognostic factor and as an indicator of the degree ofmalignancy in breast cancer.     

胃癌p16蛋白与PCNA的免疫组织化学研究

目的 :探讨p16蛋白和PCNA与胃癌发生、发展的关系。方法 :应用免疫组化LSAB法对 10 4例胃癌进行了p16蛋白和PCNA的检测。结果 :正常胃粘膜、不典型增生及胃癌均有程度不同的p16蛋白的表达 ,分别为 81.82 % ,4 5.4 6%及 50 .0 0 %。PCNA指数与肿瘤分化程度及临床分期有关。p16蛋白阳性者的PCNA指数低于p16蛋白阴性者 (P <0 .0 5)。结论 :p16蛋白和PCNA与胃癌的发生、发展有关 ,可能是反映胃癌生物学行为和预测预后的重要指标     

Proliferative activity of gastric cancer assessed by immunostaining for proliferating

The growth activity of 107 gastric carcinomas was assessed by immunohis-tochemical staining for formalin-fixed, paraffin-embedded tissue with a monoclonal antibody against proliferating cell nuclear antigen (PCNA). When the tumor doubling times (Tds) of 10 patients were estimated from the serum levels of carcinoembryonic antigen and carbohydrate antigen 19–9, there was an inverse correlation between the Tds and PCNA labeling index (LI) at P = 0.055. Flow-cytometric analysis was carried out by double staining for PCNA and DNA using fresh materials from 14 patients. The PCNA-positive cell fraction revealed by flow cytometry showed a good linear correlation with PCNA LI in routinely stained tissue. The LI of well-differentiated adenocarcinoma was significantly higher than that of the poorly differentiated type. When the LI was analyzed in well- or poorly differentiated adenocarcinoma, the value was significantly higher in the well-differentiated type with hepatic metastasis and in the poorly differentiated type with lymph node metastasis. © 1993 Wiley-Liss, Inc.     

p16及增殖细胞核抗原的表达与胃癌临床病理的关系

目的 :研究胃癌中p16蛋白、PCNA表达与胃癌临床病理的关系。方法 :应用免疫组化SP法对5 2例胃癌组织中p16蛋白、PCNA进行检测。结果 :5 2例胃癌组织中p16蛋白、PCNA表达阳性率分别为 4 4 .2 %、71.2 %。p16蛋白随着肿瘤分化程度的降低、恶性程度的增高和浸润深度的加深 ,其表达的阳性率逐渐降低 ,而PCNA的表达则相反。p16蛋白、PCNA表达与胃癌预后有关。结论 :p16蛋白、PC NA的检测可作为判断胃癌恶性程度、预测肿瘤转移和预后的参考指标     

Clinical significance of MUC1 and E-cadherin expression, cellular proliferation, and angiogenesis at the deepest invasive portion of colorectal cancer

We examined MUC1 and E-cadherin expression, cellular proliferation, and tumor vascularization at the deepest invasive portion of colorectal cancer in relation to prognosis. One hundred and ten surgically resected specimens of advanced colorectal carcinoma were studied. E-cadherin and MUC1 expression and Ki-67 labeling index (Ki-67 LI) were examined immunohistochemically at the site of deepest tumor invasion. Tumor vascularization was also examined immunohistochemically using anti-CD34 antibody to determine the microvessel count (MVC). In curative resection, patients with a high Ki-67 LI, reduced E-cadherin expression, MUC1-positive and high MVC lesion showed a significantly poorer prognosis than those with a low Ki-67 LI, E-cadherin normal, MUC1-negative and low MVC lesion, respectively. Furthermore, patients with both a high Ki-67 LI and MVC lesion showed a significantly poorer prognosis than those with other Ki-67 LI and MVC relations. Patients with both a MUC1-positive and E-cadherin reduced lesion showed a significantly poorer prognosis than those with both a MUC1-negative and E-cadherin normal lesion. The significant risk factors in order of poorer prognosis by the multivariate analysis among these factors including routinely used clinicopathologic factors were the high MVC, E-cadherin reduced expression, and lymph node metastasis. These findings indicate a high MVC at the site of deepest tumor invasion to be the most important predictor of colorectal cancer prognosis among the factors studied here.     

PCNA在胃粘膜肠上皮化生、异型增生和胃癌中表达及意义的研究

目的研究增殖细胞核抗原(PCNA)在胃粘膜、肠上皮化生,异型增生及胃癌中的表达.评估胃癌前病变的发展趋势,进一步探讨胃癌发生的分子生物学机制.方法应用免疫组化技术测定115例患者的PCNA表达情况.其中50例肠上皮化生、15例异型增生、50例胃癌.结果PCNA的表达从肠上皮化生、异型增生到胃癌呈递增趋势.在肠上皮化生为轻度表达,异型增生以中度表达为主,而胃癌多呈重度和过度表达.PCNA强阳性表达与胃癌分化程度,侵润深度,淋巴结转移密切相关.结论PCNA在胃粘膜不同疾病细胞核内的表达、反映了细胞增殖状态和生长速度、有助于判断肠上皮化生、异型增生的发展趋势及判断与监测胃癌的预后.     

胃癌组织中PTEN和VEGF PCNA的表达及相关性研究

目的 :通过对 PTEN和血管内皮生长因子 (vascular endothelial growth factor,VEGF)、增生细胞核抗原 (proliferating cell nuclear antigen,PCNA)在胃癌中表达的研究 ,结合临床病理特征 ,探讨 PTEN和 VEGF、 PCNA的相关性。方法 :应用免疫组织化学 S- P法检测 6 8例胃癌组织及 2 0例正常胃黏膜中 PTEN和 VEGF、 PCNA的表达。结果 :胃癌组织中 PTEN蛋白表达率为4 7.1% (33/ 6 8) ,显著低于正常胃黏膜的表达 10 0 % (2 0 / 2 0 ) (P<0 .0 1) ,与组织分化程度呈正相关(P<0 .0 1) ,与淋巴结转移呈负相关 (P<0 .0 5 )。 VEGF在胃癌中的表达率为 75 .0 % (5 1/ 6 8) ,显著高于正常胃黏膜的表达 10 .0 % (2 / 2 0 ) (P<0 .0 1) ,与癌组织浸润深度呈正相关 (P<0 .0 1) ,与淋巴结转移也呈正相关 (P<0 .0 5 ) ,与组织分化程度无关。 PCNA在胃癌中的表达率为 73.5 % (5 0 / 6 8) ,显著高于正常胃黏膜的表达 2 0 .0 % (4/ 2 0 ) ,与癌组织浸润深度呈正相关 (P<0 .0 1) ,与淋巴结转移呈正相关 (P<0 .0 5 ) ,与组织分化程度呈负相关 (P<0 .0 5 )。 PTEN在胃癌中的表达与 VEGF、PCNA呈负相关 (P<0 .0 1)。结论 :PTEN失活或蛋白表达降低与胃癌的组织分化程度及淋巴结转移密切相关 ,且与 VEGF、PCNA呈显著负相关。联合检     

Hypoxia-inducible factor 1 alpha in oral squamous cell carcinoma and its relation to prognosis

The aim of this study was to investigate the correlation between the expression of hypoxia-inducible factor 1 alpha (HIF-1 alpha) and proliferative activity in tumor cells, lymph node metastasis, as well as prognosis in patients with oral squamous cell carcinoma (OSCC). Fifty-seven biopsy specimens of OSCC were investigated for the expression of HIF-1 alpha and proliferating cell nuclear antigen (PCNA) by immunohistochemistry. None of the patients had received any prior treatments. The percentage of HIF-1 alpha immunopositive area (PHIA) was calculated using computer-assisted image analysis for quantitative assessment of HIF-1 alpha expression. The PCNA labeling index (LI) was evaluated as a proliferation marker. We found that the mean PHIA in all stages was 12.1% in the poor prognosis patients, and it was 6.4% in the good prognosis patients. There was a significant difference of PHIA between poor prognosis and good prognosis patients (P=0.0065). Furthermore, the mean PHIA in the patients who had no metastatic lymph nodes was 7.5%, while it was 11.7% in the patients who had metastatic lymph nodes. There was also a significant difference of PHIA between patients who had no metastatic lymph nodes and those who had metastatic lymph nodes (P=0.0487). On the other hand, significant correlation between PHIA and PCNA LI was not observed. These results provide the clinical data indicating that HIF-1 alpha may play an important role in lymph node metastasis and prognosis in patients with OSCC.     

The labelling index: a prognostic factor in head and neck carcinoma

The thymidine labelling index (LI), representing the percentage of cells in the DNA-synthesis phase, was measured in vitro prior to therapy in 87 patients with squamous cell carcinoma of the head and neck, who were treated between 1977 and 1982. The LI was not related to patient age, site of the tumour, clinical stage or histological grade. Overall survival was 44.5%. Univariate analysis demonstrated that survival was affected by the following factors: (1) age: patients older than 55 had a better outcome (p = 0.03); (2) site of the tumour (p = 0.005): laryngeal tumours had the best survival; (3) clinical stage (p = 0.05). Histological grade did not influence the survival (p = 0.41). Patients having a tumour LI higher than 15.5% (mean + 1 S.D.) had a significantly lower survival than patients with lower tumour LI (p = 0.008). A multivariate analysis using the Cox model showed that clinical stage and LI kept their prognostic impact with regard to survival. Finally, survival after relapse was lower in patients with a high tumour LI. These results demonstrate that a high tumour proliferation rate is an additional factor influencing the disease outcome in head and neck carcinoma. Patients with bad prognosis defined by this parameter could be offered a more energetic treatment.     

增殖细胞核抗原在胃癌中的表达

本文应用抗增殖细胞核抗原的单克隆抗体ＰＣ１０，用ＬＳＡＢ免疫组化方法对４０例胃癌标本进行研究。结果发现：胃癌不同分化程度各组中ＰＣＮＡ指数存在差异（Ｐ＜０．０２），低分化腺癌＞中分化腺癌＞高分化腺癌；淋巴结癌转移阳性组ＰＣＮＡ指数显著高于无淋巴结转移组（Ｐ＜０．０５）；癌细胞浸润达肌层、浆膜层者ＰＣＮＡ指数显著高于粘膜内癌（Ｐ＜０．０１）。结果显示：此方法是在石蜡切片上检测胃癌增殖指数的一个有效手段，并对判断胃癌患者的预后有一定作用。     

Expression of vascular endothelial growth factor in mouse tumours subjected to photodynamic therapy

The aim of this study was to define the appropriate fractionation interval between photodynamic therapy (PDT) for the enhancement of its anti-tumour effects. Tumour reoxygenation and the kinetics of tumour vascular cells following PDT were evaluated in mice by means of immunohistochemical staining for the vascular endothelial growth factor (VEGF) and the proliferating cell nuclear antigen (PCNA), respectively. The VEGF labelling indices (LIs) of the tumour cells and the PCNA LIs of the tumour vascular cells were assessed at various time intervals after PDT. The tumour cell VEGF LIs of the experimental groups at time points from 0 to 6 h after PDT were significantly higher than those of the control groups, but subsequently returned to control levels at 24 h after PDT. The vascular cell PCNA LI of the experimental group at 24 h after PDT was significantly lower than that of the control group, but returned to the control level at 48 h. These results indicated that the tumour subjected to PDT might be reoxygenated, and that the maximum damage to the tumour vasculature emerged at 24 h after PDT. We propose here that the fractionation interval between PDTs should be 24 h.     

P16蛋白在子宫内膜癌的表达及与细胞增殖活性的关系

目的：探讨P16基因蛋白在子宫内膜癌发生中的作用。方法：采用免疫组化方法对42例子宫内膜癌中P16蛋白和增殖细胞核抗原（PCNA）的表达进行观察。结果：P16蛋白在子宫内膜癌的阳性率为69．1％（29／42），显著低于其在正常子宫内膜（100％，8／8）及子宫内膜不典型增生（90．9％，20／22）中的阳性率（P＜0．05）。等级相关分析显示，随肿瘤组织学分级的增加、临床分期的升高及肿瘤浸润深度的增加，P16蛋白阳性率逐渐降低（P＜0．01）。PCNA标记指数与肿瘤分化程度及临床分期有关，而与浸润无关。P16蛋白阳性者的PCNA指数低于P16蛋白阴性者（P＜0．05）。结论：P16基因蛋白参与子宫内膜癌的发生、发展，并与部分生物学行为有关。