Sertoli cells promote proliferation of bone marrow-derived mesenchymal stem cells in co-culture.

Bone marrow-derived mesenchymal stem cells (BMSCs) are a major source for cell transplantation. The proliferative ability of BMSCs is an important determinant of the efficiency of transplant therapy. Sertoli cells are ‘‘nurse’’ cells for development of sperm cells. Our recent study showed that Sertoli cells promoted proliferation of human umbilical cord mesenchymal stem cells (hUCMSCs) in co-culture. Studies by other groups also showed that Sertoli cells promoted growth of endothelial cells and neural stem cells. In this study, we investigated the effect of Sertoli cells on proliferation of BMSCs. Our results showed that Sertoli cells in co-culture significantly enhanced proliferation of BMSCs (P <0.01). Moreover, co-culture with Sertoli cells also markedly increased mRNA and/or protein expressions of Mdm2, p-Akt and Cyclin D1, and decreased p53 expression in BMSCs (P <0.01 or <0.05). These findings indicate that Sertoli cells have the potential to enhance proliferation of BMSCs.

Small concentrations of TGF-β1 promote proliferation of bone marrow-derived mesenchymal stem cells via activation of Wnt/β-catenin pathway.

Bone marrow-derived mesenchymal stem cells (BMSCs) are the most promising seed cells for cell transplant. The proliferation of BMSCs is one of the most important determinants of the efficiency of MSC-based transplant therapy. It has been reported that transforming growth factor-β1 (TGF-β1) activates Wnt/β-catenin signaling and regulates cell proliferation. In this study, we investigated the effect of low concentrations of TGF-β1 on proliferation of BMSCs and the related mechanisms. BMSCs were treated with 0, 1, 5 and 10 ng/L recombinant mouse TGF-β1 for 12 h. Cell proliferation was measured by cell counting and MTT assay, and the proliferation-related signals p53, Mdm2, Akt1, Wnt3, phospho-Akt and β-catenin were measured by quantitative polymerase chain reaction (qPCR) and/or Western blot. Our results showed that TGF-β1 at low concentrations induced BMSC proliferation and expression of Mdm2, Akt1, phospho-Akt, Wnt3 and β-catenin, and inhibited p53 expression in dose dependent manner. Importantly, β-catenin siRNA significantly inhibited TGF-β1-induced BMSC proliferation. These findings suggest that low concentrations of TGF-β1 can stimulate proliferation of BMSCs, which is at least partially dependent on the activation of Wnt/β-catenin pathway.

A Study on Expression of P53 in Surface Epithelial Ovarian Tumours

Aims and Objectives (1) To perform and interpret p53 immunostaining on the diagnosed malignant surface epithelial ovarian tumours. (2) To correlate expression of p53 with histological type of malignancy. Materials and Methods A 2-year prospective study was done i.e., from October 2011 to September 2013 on ‘A study on expression of p53 in surface epithelial ovarian tumours’ in MGM Hospital, Warangal. All the ovarian surface epithelial tumour specimens, received in the pathology department during this period were considered. Results A total of 121 cases were studied, out of which benign tumours were the most common (64.4%), followed by malignancy (25.6%) and 12 cases(10%) of borderline malignancy. Most of the benign tumours were unilateral; the cases that showed bilateral involvement were mostly malignant. The maximum number of cases in the present study was seen in the age group of 31–60 year. The youngest patient was 16 year old and the oldest was 68 year old. Serous cyst adenoma was the most common neoplasm found and accounted for 53 cases (43.8%), followed by mucinous cystadenoma, which accounted for 21 cases (17.3%). Nine cases of serous cystadenofibroma (7.43%), 3 borderline serous tumours (2.47%) and 11 serous cystadenocarcinoma (9%) were found in the present study. Out of the 44 mucinous tumours, 21 were benign (17.3%), 3 were of borderline malignancy (2.47%) and 20 were malignant (16.5%). One case of mucinous cystadenocarcinoma was found to be associated with adenocarcinoma of ascending colon. One case of benign Brenner tumour was also found in the present study. The rate of p53 abnormalities varies with histological type, grade and stage of the tumour. P53 expression was more in malignant serous tumours as compared to the malignant mucinous tumours.

Avaliação da superexpressão da proteína p53 e das mutações no éxon 8 do gene TP53 em carcinomas mamários caninos e glândulas normais / Evaluation of immunohistochemical over expression of p53 protein and of mutations in exon 8 of Tp53 gene in canine mammary carcinomas and normal mammary glands

This study was undertaken with the aim to evaluate the p53 expression, applying the immunohistochemical technique to malignant primary mammary neoplasms histopathologically diagnosed in female dogs, and to investigate exon 8 of the Tp53 suppressor gene for mutation types by means of PCR-RFLP pattern of bands. Nineteen healthy mammary glands were used as a control group (group 1). Samples from 18 cases diagnosed with malignant mammary tumors (group 2), and the contralateral mammary glands (group 3) were collected during the UFRPE Veterinary Hospital routine. The tumors were diagnosed by histopathology and subdivided into grades of malignity. The streptavidin-biotin peroxidase method was used to analyze the immunohistochemical expression of p53, evaluated according to the location and intensity of stain. Expression of p53 protein was not observed in the samples of group 1. On the contrary, it was observed in all malignant tumors; the protein p53 was localized either only in the nucleus or in the nucleus and in the cytoplasm, in samples of group 2. In group 3, expression of p53 protein was observed in the tumors (2 samples) and in normal mammary tissues (4 samples). For the molecular analyses, genomic DNA was extracted and submitted to PCR-RFLP with the following endonuclease enzymes: AluI, BsoBI, DdeI and SmaI. The band pattern showed polymorphism between groups, but not between histological variants of tumors. This polymorphism detected mutations in the fragment studied - exon 8 of Tp53 - which could account for changes in nucleotides, located in the restriction sites of the endonuclease enzymes. In conclusion, the immunoexpression of p53 had no relationship with histological subtype or malignity grade, but it can be related to the presence of mammary tumors in female dogs. The PCR-RFLP technique can be an important tool for the study of mammary carcinogenesis in bitches because the polymorphism obtained may allow early diagnosis in tissues of mammary glands.

Este estudo foi realizado com o objetivo de avaliar a expressão da proteína p53, pela técnica de imuno-histoquímica, em neoplasmas mamários malignos em cadelas, além de investigar mutações no éxon 8 do gene supressor Tp53 por meio do padrão de bandas obtidas por PCR-RFLP. Dezenove mamas de cadelas saudáveis foram usadas como controle (Grupo 1). Amostras de 18 casos de tumores malignos (Grupo 2) e suas glândulas mamárias contralaterais (Grupo 3) foram obtidas na rotina do Hospital Veterinário da UFRPE. Os tumores foram identificados histologicamente e classificados em graus de malignidade. O método da estreptoavidina-biotina peroxidase foi utilizado para a análise da expressão de p53 por imuno-histoquímica, de acordo com a localização e intensidade da coloração. A expressão da proteína p53 não foi observada nas amostras do Grupo 1, mas foi encontrada em todas as amostras de tumores malignos (Grupo 2) seja só no núcleo, ou também no citoplasma. No Grupo 3, a expressão foi observada em quatro amostras normais e em duas que apresentavam tumor. Para a análise molecular, o DNA genômico foi extraído e submetido à PCR-RFLP com as seguintes endonucleases: AluI, BsoBI, DdeI e SmaI. O padrão de bandas foi polimórfico entre os grupos, mas não entre as variantes tumorais. Esse polimorfismo detectou mutações no fragmento estudado - éxon 8 do gene Tp53 - que podem resultar em alterações nos nucleotídeos, localizados nos sítios de restrição das enzimas. Esses achados levam a conclusão de que a imunoexpressão da p53 não tem relação com o subtipo histológico ou grau de malignidade do tumor, mas sim com a presença dos tumores no tecido mamário de cadelas. A PCR-RFLP pode ser usada como importante ferramenta para o estudo da carcinogênese mamária na cadela, possibilitando gerar diagnósticos precoces através do polimorfismo obtido com endonucleases de restrição pré-selecionadas.

The Relationship with the Protein Expression of p53,PTEN and the Three-year Survival Ratios in Osteosarcoma Patients / 实用放射学杂志

Objective To explore the protein expression of p53 and PTEN in osteosarcoma and the relationship between them and the three-year survival ratios of osteosarcoma patients.Methods The protein expression of p53 and PTEN was detected using immunohistochemical methods in 53 cases of osteosarcoma proved by surgery and pathology. Results There was significant difference between the both kind of relationship with the protein expression of PTEN and the three-year survival ratios of osteosarcoma patients who had negative expression of p53. There was no significant difference between the both kind of relationship with the protein expression of PTEN and the three-year survival ratios of osteosarcoma patients who had positive expression of p53. PTEN expression,which was analysed by logistic regression, was entered the regression model ,but p53 expression was not. Conclusion PTEN expression is more important than p53 expression in predicting the prognosis of the patient with osteosarcoma.

The Effects of Mutant p53 Protein Expression on the Pre-operationPatients with Breast Carcinoma Treated with Adriamycin Chemotherapy / 中国医师杂志

Objective To investigate the influences of mutant p53 protein expression on the resistance of breast cancer cells to adriamycin and efficiency of adriamycin chemotherapy of the pre-operative patients with breast carcinoma.Methods Mutant p53 protein expression in 9 cases of breast cancer was examined by using inmunohistochemical method and image semiquantified analysis.The response of human breast cancer cells to adriamycin was tested with MTT assay in order to judge the effects of mutant p53 protein expression on the pre-operative pateints with breast carcinoma treated with adriamycin chemotherapy.Results ⑴Mutant p53 protein expression in 7 cases of the human breast cancer samples were positive,the positive rate was 77%(7/9).⑵There was positive relationship between expression degree of mutant p53 protein(positive relative area) and resistance index(IC50:1/10 peak concentration) of human breast cancer cells to adriamycin(r=0 7956,P

The influence of EBV infection on p53 expression in gastric carcinoma patients from Tangshan area / 肿瘤研究与临床

Objective To investigate the influence of EBV infection on the expression of tumor suppressor gene p53. Methods EBER1 in 217 cases of gastric carcinoma were detected with in situ hybridization. Then, EBV positive and negative cases were selected for analysis of the expression of p53 by immunohistochemistry. Results In 217 cases of gastric carcinoma, there were 23 cases with EBER1 positive. The average area (AA), mean absorbency and integral absorbency of p53 expression were higher in EBV positive gastric carcinoma than those in EBV negative. Conclusions EBV infection of gastric carcinoma is closely related to the expression of p53 in this study.

Comparative Study on p53 Expression and Clinical Charateristics in Renal Cell Carcinoma / 대한비뇨기과학회지

PURPOSE: Mutations of the p53 gene are currently the most commonly recognized genetic mutations in human cancer. In some tumors, p53 gene mutations are associated with aggressiveness of the tumor and poor prognosis. There is wide variation in the reported incidence of p53 mutation in renal cell carcinoma, and little is known about its prognostic significance. To detect whether the expression of p53 could be correlated with clinical characteristics, we perfomed immunohistochemical stain on renal cell cancer samples. MATERIALS AND METHODS: Paraffin-embedded nephrectomized specimens collected from 25 patients were immunostained for p53 using the DO-1 monoclonal antibody. We evaluated the relation between p53 staining and known prognostic factors such as tumor size, pathologic stage, lymph nodal involvement, presence of metastasis, nuclear grade and cell types. RESULTS: Positive staining for p53 was detected in 32%(8/25). The staining for p53 was not statistically significant in relation with the prognostic factors such as pathologic stage(stageI&II; positive staining 6/18, 33%, stage III&IV; 2/7, 29%), lymph nodal involvement(positive 1/3, 33%, negative 7/22, 32%), distant metastasis(positive 2/6, 33%, negative 6/19, 32%), nuclear grade(nuclear gradeI&II; positive 5/18, 28%, nuclear grade III&IV; 3/7, 43%) and cell types(p>0.05). CONCLUSIONS: p53 gene mutations of the renal cell carcinoma are infrequent and show no relation to tumor size, pathologic stage, lymph nodal involvement, presence of metastasis, nuclear grade and tumor cell type in this study.

Association of p53 Protein Expression with Clinical Outcome in Advanced Supraglottic Cancer / 대한방사선종양학회지

PURPOSE: To determine the incidence and prognostic effect of p53 expression in patients with advanced supraglottic cancer. MATERIALS AND METHODS: Twenty-one cases of total 48 advanced supraglottic cancer patients who received postoperative adjuvant radiation therapy were evaluated by immunohistochemical staining employing p53 monoclonal antibody. RESULT: Three out of six stage III patients and four out of fifteen stage IV patients showed p53 expression without statistically significant difference (P=0.608). Five year survival rates are 93% in p53 negative, 86% in p53 positive patients and there was no significant difference (P=0.776). p53 expression does not show statistically significant correlation with primary tumor status (P=0.877), lymph node status (P=0.874) and age (P=0.64). CONCLUSION: There was no statistically significant correlation between traditionally known risk factors and p53 expression.

The Effects of Ultraviolet Radiation on Aging and p53 Expression in Human Skin / 대한피부과학회지

BACKGROUND: Long-term phototherapy can induce the changes of photoaging and it is reported that there is an increased chance of cutaneous squamous cell carcinomas in patients exposed to large amounts of UV radiation. OBJECTIVE: The main objective was to investigate the degree of photoaging and the presence of p53 mutations in normal skin in patients undergoing long-term phototherapy. METHOD: We performed hematoxylin-eosin and special stains, p53 and p21 immunohistochemical stains and polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) on the normal skin of patients subject to long-term UV therapy. RESULT: 1. The typical features of photoaging were not observed in patients undergoing long-term UV therapy. 2. In p53 immunohistochemical staining performed at 1 week after cessation of long-term PUVA treatment, the patient group with a culmulated UV dosage of more than 1,000J/cm2 demonstrated an increased number of p53 positive epidermal cells compared to exposed as well as unexposed normal skins. 3. The patterns of p21 immunohistochemical staining performed at 1 week after cessation of long-term PUVA and UVB treatments were similar to that of p53 immunohistochemical staining performed at 1 week after cessation of phototherapy. 4. In p53 immunohistochemical staining performed at 4 months after cessation of UV treatment, the number of p53 positive epidermal cells decreased significantly compared to that of p53 positive epidermal cells found at 1 week after cessation of UV treatment. 5. The mutation of p53 genes was not found in PCR-SSCP analysis of biopsied skins done at 1 week after cessation of long-term PUVA and UVB treatment. CONCLUSION: Long-term phototherapy did not induce the typical changes of photoaging and p53 overexpression in the epidermis of UV treated skin was a reactive process. Therefore, UV therapy can be a relatively safe treatment modality, although a closer observation for cutaneous malignancy is warrented in the patients whose cumulated UV dosage is much higher than 1,000J/cm2.

Expression of p53 gene in hepatocellular carcinomas induced by aflatoxin B1 with or without human hepatitis B virus in tree shrews

Using tree shrew as an animal model, our previous studies have demonstrated synergistic effects of aflatoxin B-1 (AFB(1)) and human hepatitis B virus (HHBV) in the induction of hepatocellular carcinoma (HCC). In the present study, we have examined expression of p53 gene in HCCs induced by AFB(1) with or without HHBV infection in tree shrews. Avidin-biotin-peroxidase complex immunohistochemical method with human p53-CM1 polyclonal antibody has been used to detect p53 expression in serial sections of paraffin-embedded liver and HCC tissues. Five out of 9 animals with HCCs (55.6%) induced by AFB(1) with HHBV infection and 2/3 animals with HCCs (66.7%) induced by AFB(1) alone expressed the p53 protein. Out of 18 HCCs examined, expression of p53 protein was observed in 9/10 moderately and poorly differentiated HCCs (0/8). None of the well differentiated HCCs (0/8) expressed p53 (0%). Similarly, no p53 expression was observed in either non-tumorous or hyperplastic liver tissues or nodules. These results suggest that p53 expression associated with p53 mutation is a late event occurring probably during tumor progression in AFB(1) and HHBV induced hepatocarcinogenesis in the tree shrew. This report is the first example of an experimental animal model where combination of human HBV and AFB(1)-induced HCCs demonstrate p53 expression.

Expression of Mutant p53 in Squamous Cell Carcinoma of Head and Neck / 대한이비인후과학회지

BACKGROUND: Expression of mutant p53 has been observed in a variety of human cancers, including head and neck squamous cell carcinoma and carcinoma of breast, colon, esophagus, lung, stomach, liver, thyroid, etc. OBJECTIVES: To establish expression frequency of p53 and correlation between p53 expression and clinicopathologic data in squamous cell carcinoma of head and neck. MATERIALS AND METHODS: Fresh tissue samples were obtained from 66 patients with squamous cell carcinoma of head and neck undergoing biopsy or surgery. Expression of mutant p53 was evaluated by immunohistochemical staining with anti-p53 monoclonal antibody in 66 paraffin-embedded tissues of squamous cell carcinoma in head and neck. The studies consisted of 13 cases of oral carcinoma including tongue, 10 cases of pharyngeal carcinoma and 43 cases of laryngeal carcinoma. RESULTS: 1) The frequency of p53 expression in squamous cell carcinoma of head and neck was 48.5%((31/66). 2) The frequency of p53 expression by tumor site was 42.6%(6/13) in oral cavity, 60%(6/10) in pharynx and 44.2%(19/43) in larynx. 3) A positive relationship was seen between p53 expression and lymphnode metastasis, representing 69.2% p53 expression in metastasis group and 16.7% p53 in non-metastasis group. CONCLUSION: Author was suggested that p53 expression in squamous cell carcinoma of head and neck was related to tumor progression and metastasis.

p53 protein expression in extrahepatic bile duct cancer

p53 mutations, a tumor suppressor gene located on chromosome 17p, are the most common genetic alterations found in human cancers. Although the p53 expression or mutation has been investigated in a variety of cancers there have been very few studies in extrahepatic bile duct cancers. In this study, we investigated the immunohistochemical expression of p53 in formalin fixed paraffin embedded archival specimens of 36 extrahepatic bile duct cancers in which p53 expression was found in eighteen (50%) cases. There was no significant difference in age, gender, size of tumor, histologic grade, extent of tumor involvement, lymph node metastasis and tumor resectability according to p53 immunoreactivity. Comparison of survival duration according to p53 expression showed no significant difference. In conclusion, we reported 50 percent of p53 expression in extrahepatic bile duct cancers by immunohistochemical staining and we found no prognostic significance of p53 expression in dinicopathologic parameters.

Immunohistochemical Detection of p53 Gene Mutation in Urine Samples in the Patients with Bladder Cancer / 대한세포병리학회지

Although bladder cancers are very common, little is known about their molecular pathogenesis. It is known, that p53 alteration is found in about 60%p of muscleinvasive bladder cancer, necessiating aggressive therapy and poor outcome. We examined the nuclear expression of p53 protein, using D07 monoclonal antibody in the urine samples, from 31 patients with transitional cell carcinoma of the bladder to investigate the correlation of p53 overexpression with histologic grades and depth of invasion. The positive rate of p53 protein was 27%o in superficial bladder tumor, but increased up to 71% in the invasive bladder carcinomas. The overexpression of p53 protein increased according to Mostofi grading system from 18% in grade I, 45% in grade Il, and up to 100% in grade ill. The p53 expression tended to be higher in the invasive and high grade bladder cancers than in the superficial and low grade ones(p<0.05). These results suggest that immunohistochemical analysis of the urine specimen in the bladder cancer patients could be a useful method of screening for the presence of p53 mutant protein. The mutant p53 protein expression may be an indicator of bladder cancer with more proliferative potential and/or aggressive biologic behavior.

Microsatellite instability of chromosome 17p and mutant P53 protein expression in hepatocellular carcinoma / 第三军医大学学报

0 05). However, P53 positive rate in AFP positive HCC was significantly higher than that in AFP negative HCC ( P