Loss of maspin expression is associated with development and progression of gastric carcinoma with p53 abnormality

Maspin, a serine protease inhibitor related to the serpin family, was originally identified in normal mammary epithelium. Reduced expression of maspin is related with development, invasion and metastasis of certain human cancers. In the present study, the expression of maspin was examined in gastric mucosa, adenoma and carcinoma by immunohistochemistry and RT-PCR. In non-neoplastic mucosa, maspin was expressed in cytoplasm and cell membrane of foveolar epithelia, fundic glandular cells and pyloric glandular cells. Maspin expression was lost in 71% (71/100) of gastric carcinomas, and in 19% (4/21) of adenomas, respectively. Loss of maspin expression was significantly associated with poorly differentiated histology, advanced stage and deep invasion (P<0.001). There was an inverse correlation between maspin expression and abnormal p53 accumulation. Maspin mRNA expression was lost in all of 8 gastric carcinoma cell lines that was retrieved after treatment with demethylation agent 5-aza-2'-deoxycytidine in 5 of 8 cell lines. These results suggest that loss of maspin expression partly due to DNA methylation may participate in tumor development and progression of gastric carcinoma in relation with p53 pathway. Loss of maspin expression may serve as a biological marker of high-grade malignancy.     

Cx43、Skp2在卵巢上皮性肿瘤组织中的表达及临床意义

背景与目的:间隙连接蛋白43(connexin43,Cx43)是间隙连接蛋白家族的主要成员,在多种肿瘤细胞中表达下降。它在许多细胞系以依赖间隙连接(Gapjunction)的途径发挥抑癌作用。最近研究发现它还可能通过抑制S期激酶相关蛋白2(S-phase kinase associated protein2,Skp2)的表达而起到抑制肿瘤生长的作用。Skp2是F鄄box蛋白家族的成员,它能特异性识别并促进某些调节G1期进程的关键性细胞周期调节物的降解,在许多肿瘤中表达升高。本研究检测Cx43和Skp2在卵巢上皮性肿瘤中的表达,探讨它们的表达与卵巢癌发展的关系,以及这两种蛋白表达的相互关系。方法:收集81例卵巢上皮性肿瘤的外科手术石蜡标本(良性13例、交界性12例、恶性56例),用免疫组化的方法检测Cx43和Skp2蛋白的表达水平。分析它们的表达水平与临床病理参数的关系以及二者的相互关系。结果:Cx43蛋白在卵巢上皮性良性、交界性及恶性肿瘤中阳性率分别为84.6%、66.7%和33.9%,经免疫组化半定量分析,其在上皮性卵巢癌中的表达水平明显低于良性肿瘤(P<0.01)和交界性肿瘤(P<0.01),在不同年龄及组织类型的卵巢癌中表达无显著性差异(P>0.05),而在中低度分化组、Ⅲ～Ⅳ期组及有淋巴结转移组分别明显低于高分化组(P<0.05)、Ⅰ～Ⅱ期组(P<0.05)及无淋巴结转移组(P<0.0     

Maspin,VEGF and p53 Expression in Small Biopsies of Primary Advanced Lung Cancer and Relationship with Clinicopathologic Parameters

Maspin, one of the serine protease inhibitors, has been shown to inhibit tumor progression and metastasis. We aimed to investigate maspin, p53 and VEGF expression in patients with squamous cell carcinoma (SCC), adenocarcinoma (AC) and small cell lung carcinoma (SCLC). The study included 28 SCC, 18AC, 17 SCLC biopsy samples. We used the streptavidin biotin immunoperoxidase method to test for maspin, p53 and VEGF antibodies. Medical records of these patients were reviewed from archival files. Cytoplasmic maspin expression was detected in 89.3%, 77.8%, 52.9% of SCC, AC and SCLC, respectively. The rate was significantly higher in non-small cell lung cancer (NSCLC) and SCC than SCLC (p = 0.013, p = 0.021, respectively). The mean percentages of maspin expression were significantly higher in NSCLC, SCC and AC than in SCLC (p = 0.0001, p = 0.0001, p = 0.038, respectively). In ACs, maspin and p53 expressions were correlated, although this was not statistically significant (p = 0.053, r = 0.464), and maspin positive cases had a significantly higher T status compared to negative cases (p = 0.036). In SCC, the stage of disease was positively correlated with p53 (p = 0.007, r = 0.536) and negatively correlated with VEGF expression (p = 0.013, r = −0.498). Multivariate analysis demonstrated that stage of disease was a significant independent prognostic parameter in NSCLC (95% confidence interval: 1.067–3.969; p = 0.031). Although maspin expression is higher in SCC and AC, and is related with higher T status in AC, our data did not indicate its prognostic significance. Larger scale studies are needed to reveal the exact role of maspin in lung cancer pathogenesis.     

The correlation between IGF-II and Bcl-2 expression in colorectal adenocarcinoma

Our aim for this study was to investigate the correlation and clinical significance between the expression of IGF-II and Bcl-2 in colorectal adenocarcinoma, especially in terms of the metastasis of colorectal adenocarcinoma. Sixty paraffin embedded samples of colorectal adenocarcinoma were selected, and fifteen normal colorectal tissues were used as controls. IGF-II mRNA was detected using in situ hybridization, and the expression of Bcl-2 along with the proliferating cell nuclear antigen (PCNA) protein was detected through immunohistochemistry. The TUNEL assay was used to detect apoptosis. Specimens with a positive cell ratio less than 30% were defined as negative. The levels of IGF-II mRNA and the Bcl-2 protein were significantly higher in colorectal adenocarcinoma (39.64 ± 7.38% and 30.74 ± 7.22%, respectively) than in normal colorectal tissues (22.56 ± 4.21% and 12.17 ± 1.94%, respectively) (P < 0.01). The levels were related to Dukes′ stage and lymph node metastases, but were unrelated to patient age, gender, tumor site, tumor size, and tumor differentiation. Also, a negative correlation was observed between IGF-II mRNA and Bcl-2 protein (P < 0.05) during Dukes′ stages. In addition, a positive correlation between IGF-II mRNA and PCNA or apoptosis, as well as a negative correlation between Bcl-2 and apoptosis were observed (P < 0.01). There was no correlation between Bcl-2 and PCNA (P > 0.05). The patients detected as IGF-II mRNA (+) and Bcl-2 (−) showed the worst prognosis. The expression of IGF-II and Bcl-2 was correlated with the clinical manifestation of colorectal adenocarcinoma; thus, the assessment of both IGF-II and Bcl-2’s status will provide important information regarding the diagnosis and prognosis of colorectal adenocarcinoma.     

Serum p53 antibody as a useful marker for monitoring of treatment of superficial colorectal adenocarcinoma after endoscopic resection

Background. Mutation of the p53 gene is a genetic alteration found in human cancers. Overexpression of p53 has been found to induce antibody production in serum, and, recently, the simple detection of serum antibody has been made possible. The aim of this study was to evaluate the potential role of serum p53 antibody in the early diagnosis of superficial colorectal cancer and in the monitoring of its treatment after endoscopic resection. Methods. In a prospective study, our subjects were 27 patients with superficial colorectal adenocarcinomas, whose results were compared with those in 38 patients with benign adenomas; all patients were treated by endoscopic resection. The correlation between serum p53 antibody levels before and within 3 weeks after resection was determined, using an immunoassay. Immunohistological staining for p53 was also performed, and its sensitivity was compared with that of two other tumor markers. Results. Preoperatively, serum p53 antibody was detected in 63.0% (17/27) patients with adenocarcinoma and in 2.6% (1/38) patients with adenoma, showing a significant difference (P < 0.001). However, the two other markers carcinoembryonic antigen (CEA) and carbohydrate antigen CA19-9, showed no significant difference between superficial colorectal adenocarcinoma and adenoma. The serum p53 antibody status was strongly correlated with p53 immunostaining in adenocarcinoma (P = 0.0065), but there was no significant correlation in adenoma (P = 0.973). Sixteen (94.1%) of 17 seropositive adenocarcinoma patients, showed negative conversion after complete tumor resection, and all these 16 patients remained seronegative. Conclusion. The detection of serum p53 antibody is expected to serve as a new genetic marker, determined by serological analyses, for aiding in the early diagnosis of superficial colorectal cancer and indicating its local curability after endoscopic treatment. Received: February 21, 2000 / Accepted: September 28, 2000     

Maspin gene expression is a significant prognostic factor in resected non-small cell lung cancer (NSCLC). Maspin in NSCLC

Maspin is a member of the serpin (serine protease inhibitor) superfamily, and some experimental studies revealed a potential tumor suppressor activity of maspin. To reveal clinical significance of maspin status in non-small cell lung cancer (NSCLC), we quantitatively evaluated maspin gene expression in lung primary tumors cut from a total of 55 resected NSCLC patients. Maspin expression in squamous cell carcinoma (Sq) was significantly higher than that in adenocarcinoma (Ad, p=0.011). Five-year overall survival rates of maspin-high and maspin-low patients were 67.7 and 41.4%, respectively, demonstrating a significant favorable prognosis of maspin-high patients (log-rank, p=0.042). A multivariate analysis confirmed that high maspin expression was an independent and significant factor to predict a favorable overall survival (p=0.031). These results suggested that maspin expression was significantly increased in Sq than in Ad, and that increased maspin expression was a significant factor to predict a favorable prognosis in resected NSCLC.     

Maspin expression in early oral tongue cancer and its relation to expression of mutant-type p53 and vascular endothelial growth factor (VEGF)

Even though oral tongue cancer is generally diagnosed at an early stage, the prognosis is poor due to frequent recurrence. Therefore, it is important to identify factors predictive of recurrence and to treat aggressively those patients with a high probability of recurrence. The relationship between angiogenesis and recurrence in tongue cancer has been widely investigated but no consensus has been reached. Mutant-type p53 and VEGF are known to be related to angiogenesis, and maspin is a potent angiogenic inhibitor but its role in tongue cancer has scarcely been examined. We observed the expression of maspin, mutant-type p53 and VEGF by immunohistochemistry in 33 patients with stages I and II oral tongue cancer. And the relationships between maspin, mutant-type p53, VEGF expression and recurrence were analyzed. Maspin and VEGF displayed a cytoplasmic staining pattern and mutant-type p53 a nuclear pattern. None of expression of maspin, mutant-type p53, and VEGF was significantly correlated with tumor recurrence (p=0.34, 0.56, and 0.33, respectively) and survival. Maspin expression was negatively correlated with both mutant-type p53 expression (p=0.02), and VEGF expression (p=0.01). There was no correlation between age, sex, clinical staging, and recurrence. In conclusion, the expression of maspin is not related to recurrence of early stage oral tongue cancer. It is inversely correlated with that of mutant-type p53 and of VEGF, suggesting that the maspin gene is a mutant-type p53 target in vivo and may contribute to regulate VEGF expression.     

Aberrant maspin expression in human endometrial cancer

Maspin, a mammary serine protease inhibitor, was originally reported as a tumor suppressor gene in breast cancer. The purpose of the present study was to examine maspin expression and evaluate its clinicopathological significance in endometrial cancer. We examined maspin expression immunohistochemically in 41 cases with endometrioid adenocarcinoma. DNA methylation status at the maspin promoter region was determined by the methylation-specific polymerase chain reaction method. Aberrant maspin expression was observed in 27 (66%) of 41 endometrioid adenocarcinomas but not in normal endometrial glands. Maspin immunoreactivity of the tumor cells varied in incidence and density among tumors. Positive staining was correlated significantly with the presence of squamous differentiation (presence vs absence = 11/11 [100%] vs 16/30 [53%], P < 0.05), and nuclear subcellular localization of maspin protein was also significantly associated with squamous differentiation (nuclear positive vs nuclear negative = 6/11 [54%] vs 2/30 [6.7%], P < 0.05). An inverse correlation between their immunoreactivity and methylation status was observed (P < 0.01). Three of the four cell lines established from endometrioid adenocarcinomas overexpressed maspin mRNA and its protein product. In a maspin-negative cell line, maspin expression was induced by treatment with 5-aza-2'-deoxycytidine, a DNA demethylating agent. There was no significant correlation between maspin expression and any clinicopathlogical data. These findings suggest that maspin induced by DNA demethylation at the promoter region may contribute to squamous differentiation of tumor cells in endometrioid adenocarcinomas.     

Clinicopathological significance and molecular regulation of maspin expression in ductal adenocarcinoma of the pancreas

We evaluated the biological relevance of maspin expression in pancreatic ductal adenocarcinoma and studied regulatory mechanisms of maspin gene activation in pancreatic carcinoma cell lines. Maspin expression was immunohistochemically detected in a series of 57 pancreatic ductal adenocarcinomas, 51 (90%) of which were classified as high-expressers. In lymph node metastases, maspin expression was somewhat decreasingly found in 39/49 (80%). Maspin high-expressers showed predominantly a low histological grade (p=0.013). Moreover, maspin expression was found in two mixed ductal-endocrine carcinomas, but not in 10 endocrine tumors and the surrounding normal pancreatic tissues. Using a luciferase reporter system, maspin promoter activity was induced in the maspin-positive pancreatic cancer cell lines as well as maspin-negative PANC-1 cells. Additionally, treatment with the DNA methyltransferase inhibitor, 5-aza-2' deoxycytidine, and histone deacetylase inhibitor, trichostatin A, led to re-expression of maspin mRNA in PANC-1 cells. Our results indicate that maspin expression is up-regulated in most if not all pancreatic ductal adenocarcinomas and may be related to the development and differentiation, and that DNA methylation and histone deacetylation may suppress maspin gene activation in pancreatic cancer cells.     

Maspin expression in adenocarcinoma of the ampulla of Vater: relation with clinicopathological parameters and apoptosis

AIM: Maspin is a unique serine proteinase inhibitor which has tumor suppressor activity. The aim of the present study was to investigate the role of maspin in ampullary adenocarcinomas, its correlation with apoptosis and its value as a prognostic marker. PATIENTS AND METHODS: Twenty-three cases of ampulla of Vater adenocarcinoma were collected from archival material. For each sample, maspin, M30, p53 and Mib1 immunohistochemical reactivity were evaluated and results compared with clinicopathological parameters. RESULTS: A statistical relation was found between nuclear maspin and M30 (Spearman's Q = 0.46, p = 0.02), and p53 (Kruskal-Wallis = 0.03); a trend was found between nuclear maspin and pT (Kruskal-Wallis = 0.09), and pM (Mann-Whitney = 0.08) and pN status (Fisher's mid-point test: p = 0.070). CONCLUSION: The present study evaluated the role of maspin in ampullary adenocarcinomas and for the first time demonstrated its association with apoptosis, tumor growth and metastasis.     

Using a combination of cytochrome P450 1B1 and β-catenin for early diagnosis and prevention of colorectal cancer

Background: Although fecal occult blood test and invasive endoscopic examination are common used to detect colorectal adenomas and cancers, non-invasive and specific biomarkers are still under investigation. The objective is to evaluate the biomarker CYP1B1 alone or in combination with aryl hydrocarbon receptor (AhR), nuclear β-catenin, p53 or bcl-2 for early diagnosis and prevention of colorectal cancer. Methods: These biomarkers were analyzed semi-quantified across 231 colonic tissues including 97 adenocarcinomas, 85 adenomas and 49 non-neoplastic colons using immunohistochemistry. In order to differentiate non-neoplastic colons from colorectal neoplasms (adenoma and carcinoma), the values for CYP1B1, AhR, nuclear β-catenin, p53 and bcl-2 expressions were subjected to discrimination analysis, then the cross-validation, sensitivity and specificity of these models were calculated. Results: Expressions of CYP1B1, p53, nuclear β-catenin and bcl-2 were significantly associated with colorectal carcinogenesis (p < 0.01 for the trend test). The overexpression rates for CYP1B1, p53, nuclear β-catenin and bcl-2 were significantly higher in the adenoma and carcinoma groups than in the non-neoplastic colon group (p < 0.05). The discrimination models showed that a combination of two biomarkers was better than a single biomarker, and provided specificity ranging from 39% to 100% and sensitivity ranging from 43% to 82% for colorectal carcinoma. Conclusions: The increase in expression of CYP1B1 occurred not only in colorectal carcinoma and but also in adenoma. Moreover, a screening panel of CYP1B1 in combination with nuclear β-catenin was the most suitable marker pair to screen for colorectal carcinoma based on this study.     

Maspin in thyroid cancer: its relationship with p53 and clinical outcome

The serine protease inhibitor maspin has been reported to inhibit invasiveness and motility of tumor cells. Additionally, a p53-dependent regulatory pathway of maspin in human cancer has been indicated. In a pre-study we were able to detect maspin protein in papillary thyroid carcinomas (PTC), whereas normal (tumor-free) thyroid tissue, follicular adenomas, follicular carcinomas, poorly differentiated carcinomas and undifferentiated carcinomas of the thyroid were maspin-negative. The first aim of our study was to determine the prognostic value of maspin protein expression for the recurrence-free and overall survival of PTC patients undergoing radical thyroidectomy and postoperative irradiation. Secondly, maspin expression was correlated to p53 protein expression in order to gain additional information on a possible regulatory influence of the wild-type p53 protein on maspin. An immunohistochemical approach study was performed on 68 tumor specimens. Maspin protein expression was detectable in 48 of 68 patients (71%; M+). After a median follow-up of 81 (26-117) months the median recurrence-free survival was 60 (28-117) months for M+ and 42 (11-108) months for M- (p=0.03). After 110 months 83% of patients had recurrence-free disease in M+, whereas in M- only 40% of patients were recurrence-free. The median long-term survival was 81 (42-108) months for M+ and 55 (21-99) months for M- (p=0.03). After 5 years, M+ and M- patients had a total survival of 98 and 80%, and after 9 years 90 and 60%, respectively. Mutant-type p53 expression was detectable in 17 of 68 PTC (25%). Mt p53 was positive in 1 of 47 M+ (2%) compared with 16 of 20 M- (80%, p<0.01). This study indicates that maspin protein possibly functions as a clinically relevant inhibitor of tumor progression, preventing local invasiveness and further systemic progression of papillary thyroid carcinomas. Our data hint of a p53-dependent regulatory pathway of the maspin protein in human cancer.     

Expression of Maspin in Non–Small-Cell Lung Cancer: Correlation with Clinical Features

PurposeMaspin is a member of the serpin (serine protease inhibitor) family and has been shown to be a suppressor of tumor growth and metastasis in several types of tumors. The objective of this study was to evaluate whether maspin is a prognostic factor in patients with non–small-cell lung cancer (NSCLC).Patients and MethodsWe investigated maspin expression in 181 patients with curatively resected NSCLC by means of immunohistochemistry. We also determined whether expression of maspin correlates with the microvessel density (MVD) level.ResultsThe incidence of strong maspin expression in patients with squamous cell carcinoma was significantly higher than that in patients with other histology (46 of 70 [65.7%]; P < .0001). There was no significant difference between maspin expression status and MVD. Prognosis was defined as progression-free survival (PFS) and overall survival (OS). There was no difference in PFS or OS between patients with strong and weak maspin expression among all patients. However, for squamous cell carcinoma, the PFS and OS rates for patients with strong maspin expression were significantly higher than those for patients with weak maspin expression (PFS, P = .004; OS, P = .001). In multivariate analysis on squamous cell carcinoma, strong maspin expression was an independent favorable prognostic indicator (PFS, P = .03; OS, P = .01).ConclusionStrong maspin expression was an independent factor in predicting a favorable prognosis in squamous cell carcinoma of lung.     

胃癌组织中Maspin 蛋白与血管内皮生长因子-C的表达及其相关性

 目的 探讨胃癌组织中Maspin蛋白与血管内皮生长因子-C（VEGF-C）的表达及其相关性。方法 收集哈尔滨医科大学附属肿瘤医院2002年～2003年间手术切除的胃癌组织石蜡标本61例,采用免疫组化技术检测胃癌组织中Maspin蛋白与血管内皮生长因子-C（VEGF-C）的表达情况,结合临床病理特征进行分析,并探讨两者蛋白表达水平的相关性。结果 Maspin蛋白的阳性表达率为50.8%（31/61）。淋巴结转移阳性组织中Maspin蛋白的阳性表达率明显低于淋巴结转移阴性的组织（32.6%vs94.4%,P=0.000）。且Maspin蛋白在组织分化较低,TNM分期较晚,周围有肿瘤浸润的病例中表达率显著降低（P分别为0.018,0.011和0.028）。VEGF-C在胃癌组织中的阳性表达率为86.9%（53/61）。淋巴结转移阳性组织中VEGF-C蛋白的阳性表达率明显高于淋巴结转移阴性的组织（95.3%vs66.7%,P=0.006）。Maspin蛋白与VEGF-C在胃癌组织的表达存在负相关（Spearmanr=-0.429,P〈0.05）。结论 Maspin蛋白在胃癌组织中低表达,胃癌的浸润转移能力增强可能与Maspin蛋白表达下调、缺失相关;VEGF-C的高表达与肿瘤淋巴结转移关系密切;Maspin蛋白与VEGF-C在胃癌组织中的表达呈明显负相关。     

Correlation of Skp2 with carcinogenesis, invasion, metastasis, and prognosis in colorectal tumors

In colorectal tumors, S-phase kinase-associated protein 2 (Skp2) still has numerous important questions unanswered: its expression in adenomas, its correlation with key clinicopathological indices, its association with patient prognosis, its variation in lymph node metastases, and its association with many cell-cycle regulators. To answer these questions in colorectal tumors, Skp2, cyclin A, cyclin B1, cyclin E, CDK2, and Ki67 were immunohistochemically stained in 12 normal mucosa, 36 adenomas, 11 carcinomas in adenomas, 102 primary carcinomas, and 12 paired lymph node metastases; and Skp2 was examined by Western blot in 8 pairs of normal mucosa and carcinomas. Situated in nuclei, Skp2 expression significantly increased from normal mucosa through adenoma to primary carcinoma (p<0.0001), from mild through moderate to severe dysplasia in adenomas (p=0.038), from peripheral adenoma to paired central carcinoma (p=0.0033), and from primary carcinoma to lymph node metastasis (p=0.015), and these increases were confirmed by Western blot. Expression, however, relatively declined significantly in the primary carcinomas showing deep invasion (p=0.0113), lymph nodal metastases (p=0.0268), and poor prognosis for all (p=0.0104) or stage III patients (p=0.0119). High Skp2 was also significantly linked with elevated cyclin A, cyclin B1, cyclin E, CDK2 (in primary carcinomas only), and Ki67 in both adenomas and primary carcinomas. Thus, overexpression of Skp2 is associated with colorectal carcinogenesis and late metastasis to lymph nodes, whereas relative reduction of Skp2 is correlated with local invasion of primary carcinoma.     

Maspin在非小细胞肺癌中的表达及与p53的相关性

目的探讨maspin在非小细胞肺癌中的表达及其与p53的关系。方法对99例非小细胞肺癌组织maspin及p53蛋白的表达情况通过免疫组化法进行检测,分析其临床病理意义及相关性。结果 maspin蛋白表达与肿瘤病理类型、肿瘤分化程度、淋巴结状态及临床分期相关（P〈0.05）。而p53蛋白的表达则与临床分期、淋巴结状态相关（P〈0.05）。maspin与p53蛋白的表达呈负相关（γ=-0.180）,但无统计学意义（P=0.075）。结论 Maspin及p53蛋白的表达在非小细胞肺癌的发生、发展过程中起重要作用,但本研究未发现两者具有统计学意义的相关性。