p53 gene mutation and numb expression in colorectal cancer and its clinical significance / 中华普通外科杂志

Objective:to study the mutation of p53 gene in colorectal cancer, analyze the relationship between p53 gene mutation and numb expression pattern, and explore its clinicopathological significance in colorectal cancer.Methods:p53 gene mutation in 60 colorectal cancer tissues was analyzed by polymerase chain reaction (PCR) and DNA sequencing, and the expression of numb protein was detected by Western blot. The colon cancer cell lines HCT116 (+), HCT116 (-) and flow cytometry were used. The survival curve was drawn by Kaplan Meier method.Results:p53 gene mutation was found in 31 of 60 tissues (52%), and the mutation times of exons (E) 5, 6, 7 and 8 were 5, 6, 12 and 11 respectively. The expression level of numb in p53 mutation group was significantly lower than that in non mutation group ( P=0.009). The prognosis of patients with low expression of numb (39 cases) was worse than that of high expression of numb (21 cases) ( P=0.015). Its expression level is closely related to the degree of differentiation, lymph node metastasis and TNM stage (all P<0.05). After the two cell lines were transferred into numb, the cell cycle appeared G2-M phase arrest and proliferation was inhibited, while dapt had G1-S phase arrest. Conclusion:p53 gene mutation related to the expression of numb in colon cancer, which has significant effect on the prognosis.

Histopathological Analysis of Esophageal Mucosa in Patients with Achalasia.

Background/Aims: Achalasia is an esophageal motor disorder that leads to functional esophageal obstruction. Food stasis and bacterial fermentation can predispose an individual to esophageal mucosal inflammation, causing multifocal dysplasia and increasing the risk of developing esophageal squamous cell carcinoma. We aimed to evaluate esophageal mucosal alterations in achalasia patients and determine clinical factors associated with the histopathological findings. Methods: From 2009 to 2013, we obtained endoscopic biopsies from the lower and middle esophagus of 22 patients with achalasia and 17 controls. Patients' clinical data and histological severity of esophagitis were retrospectively analyzed. Additionally, immunohistochemical staining for CD3, CD20, Ki-67, and p53 was conducted. Results: The median age of achalasia patients was 49.5 years (range, 27 to 82 years), and there were nine males (40.9%). The median symptom duration was 5.8 years (range, 1 to 33.5 years), and 10 patients (45%) underwent previous treatment (nine, balloon dilation; one, botulinum toxin injection). Achalasia patients had significantly more severe esophagitis than did controls (p=0.001, lower esophagus; p=0.008, middle esophagus), and the number of CD3-positive lymphocytes exceeded that of CD20-positive lymphocytes (p<0.001). Achalasia patients also had a higher esophageal Ki-67 proliferation index (p=0.048). Although statistically nonsignificant, p53 expression was only observed in achalasia patients. There was no association between the histological severity of esophagitis and other clinicopathological findings. Conclusions: Achalasia patients showed significantly severe histological esophagitis and a high Ki-67 proliferation index, indicating an increased risk of neoplastic progression. Therefore, careful endoscopic inspection is necessary for the early detection of superficial neoplasia in these patients.

Progress of cell phenotypes and molecular subtypes as well as p53 gene abnormalities in the prognosis of diffuse large B-cell lymphoma / 肿瘤研究与临床

Diffuse large B-cell lymphoma (DLBCL) is a group of heterogeneous diseases, and further classification is of great value for prognostic stratification. In recent years, some subtypes with poor prognosis have been recognized, such as "double-hit" lymphoma or "double-expressing" lymphoma. In addition, the new molecular classification provides a new perspective for the accurate diagnosis and prognosis stratification of DLBCL. The mutation and deletion of p53 gene are high risk factors in the traditional sense of DLBCL, while with the application of the next generation sequencing and other technologies, the influence of more accurate gene mutation or deletion on prognosis and its prognostic value in new cell molecular subtypes still need to be further confirmed. This paper reviews the progress of cell phenotypes and molecular subtypes as well as p53 gene abnormalities in the prognosis of DLBCL.

Inhibitory Effect and Target Prediction of Genistein on the Growth of Human Nasopharyngeal Carcinoma CNE 1 Cells / 中国药房

OBJECTIVE：To study the inhibi tory effects of genistein on the growth of human nasopharyngeal carcinoma. CNE 1 cells and predict its potential target. METHODS ：CCK-8 method was used to test the effects of 0（blank control ），12.5，25，50， 100，150 µmol/L genistein on the proliferation of CNE 1 cells after treated for 24，48，72 h. Flow cytometry was carried out to detect the effects of 0（blank control ），15，30，60 µmol/L genistein on the cell cycle and ap optosis of CNE 1 cells after treated for 24 h. Scratch test was used to investigate the effects of 0（blank control ）， 10， 20， 30 µmol/L genistein on themigration ability of CNE 1 cells after treated for 24 h. High （No.18210156） throughput sequencing was conducted to discover the differential genes in CNE 1 cells after treated with 0（blankcontrol），30 µmol/L genistein for 24 h. RT-qPCR assay was adopted to verify the mRNA expression of related differential genes in above trials. RESULTS ： Compared with blank control，12.5，25，50，100，150 µmol/L genistein sho wed significant inhibitory effect on the proliferation of CNE 1 cells（P＜ 0.01），in a concentration- time-effect manner ；15，30 µmol/L genistein could arrest CNE 1 cell cycle at G 0/G1 stage（P＜0.05 or P＜ 0.01）；30，60 µmol/L could arrest CNE 1 cell cycle at G 2/M stage and promoted cell apoptosis （P＜0.05 or P＜0.01）. 10，20，30 µmol/L genistein could significantly inhibit the migration ability of CNE 1 cells（padj＜0.01）. High throughput sequencing revealed a total of 2 271 differentialgenes（P＜0.05），1 154 of which were up-regulated while 1 117 of which were down-regulated ；8 potential target genes ，including p53，p21，STC2，FGF2，CDK6，CYCLIN D ，PI3K，AKT，were screened by cell experiment. After validated by RT-qPCR assay ，mRNA expression of p53，p21，STC2，FGF2，CDK6，CYCLIN D and AKT were significantly down-regulated（P＜0.05），which consistent with the sequencing results. CONCLUSIONS ：Genistein can effectively inhibit the growth of human nasopharyngeal carcinoma CNE 1 cells，the mechanism of which may associated with inhibiting the expression of mutant gene p53，restoring the function of wild-type P 53 protein and inhibiting the activity of PI 3K/Akt pathway.

Identification of R249 Mutation in P53 Gene in Tumoral Tissue of Tongue Cancer.

BACKGROUND: Tongue cancer is the most common malignancy of the mouth. In recent decades, reported tongue cancer incidence and mortality rates have increased all over the world while survival has not improved that sometimes is related to mutation, especially in gene P53 (such R249, R248 mutations). Hence, this study aimed to identification of R249 mutation in P53 gene of tumor tissue in tongue cancer. METHODS: In a cross-sectional study, 48 patients with squamous cell carcinoma (SCC) of tongue were selected, and mutation of R249 was investigated in sample of tumors. In addition, demographic data and medical history of patients were determined and registered in a collected data form. Finally added data were entered to computer and analyzed by SPSS software. RESULTS: Polymerase-chain reaction test done on tissue samples from cancer patients showed that in a studied sample of 48 patients, 4 of them (8.3%) had R249 mutation. After selecting the codon 249 as a hotspot in oral cancer, forward and reverse primers for amplification of exon 7 were obtained from the articles. CONCLUSIONS: Considering the findings of our study, R249 mutation in P53 gene in patients with SCC is relatively high. Age and alcohol consumption were factors affecting incidence of the mutation. It is necessary to take an early treatment with a single lesion of tongue to prevent severe disease and prevent disease in patient's family with screening test and prevent cancer in future with gene therapy.

Cytotoxic effects of mono- and di-rhamnolipids from Pseudomonas aeruginosa MR01 on MCF-7 human breast cancer cells.

Rhamnolipids produced by P. aeruginosa MR01 were fractionated into mono- and di-rhamnolipids, and their dominant congeners, Rha-C10-C10 and Rha-Rha-C10-C10, were shown by mass spectrometry. Minimum surface tensions and critical micelle concentrations (CMC) were determined as "≃34 mN/m; ≃26.17 mg/l;" and "≃29 mN/m; ≃29.63 mg/l" for mono- and di-rhamnolipids, respectively. Spectrophotometry measurements provided a close approximation of CMC. Contact angle and diameter of wet area were determined for rhamnolipid-containing drops on hydrophobic paper to display their capability for alteration of surface wettability. Wet area measurement is a simple, reliable method not requiring a Drop Shape Analyzer. Cell viabilities determined by MTT assay showed a decline in a dose-dependent manner and estimated IC50 values were 25.87 µg/ml and 31.00 µg/ml for mono- and di-rhamnolipids treating MCF-7 cells for 48 h. Morphological observations using the inverted phase-contrast microscopy and fluorescence microscopy via Hoechst staining revealed the apoptotic characteristics in treated MCF-7 cells. The semi-quantitative RT-PCR method demonstrated that expression of the p53 gene in mRNA levels significantly (P < 0.05) increased when treated with 30 µg/ml of each rhamnolipid compound for 12 h. It can be concluded that rhamnolipids derived from MR01 show significant anticancer potential against MCF-7 cell line and should be further investigated as natural, therapeutic anti-tumor agents.

Prognostic value of the Epstein-Barr virus and tumor suppressor gene p53 gene in nasopharyngeal squamous cell carcinoma.

AIMS AND METHODS: Retrospectively, this paper compared the differences of the Epstein-Barr virus (EBV)-encoded small RNAs (EBERs), protein expression and gene mutations of tumor suppressor gene p53 (TP53) in keratinized nasopharyngeal squamous cell carcinoma (KNSCC) and nonKNSCC, and the relationships between pathological features and the prognosis of patients were analyzed. RESULTS: The positive rate of EBERs hybridization and TP53 expressions was 76.3% and 52.2%, respectively, while the mutation rate of TP53 gene was 39.6%. Logistic regression analysis showed direct relationships between the subtypes of nasopharyngeal squamous cell carcinoma (NPSCC) and EBERs-positive, or frequent consumption of pickled food. Overall survival rates of patients with positive TP53 expression, the TP53 gene mutations, vascular invasions, organ metastases, lymph node metastasis, and clinical recurrence were significantly lower than those of patients without those symptoms. The poorer prognosis was related to regularly drinking and the advanced age. According to the Cox regression analysis, we found that the main prognostic factors of NPSCC patients were the aging, recurrence, TP53 gene mutations, especially exon 7 or 8 mutations. CONCLUSIONS: We concluded that there were the correlations between NPSCC subtypes with EBV infection and frequent intaking of pickled food, while aging, clinical recurrence, and TP53 gene mutations were independent predictors for the poor prognosis of nasopharyngeal carcinoma.

Distribution of epidermal growth factor receptor, bone morphogenetic protein-2, and p53 in kidney of healthy newborn, adult and old highland-plateau yaks.

BACKGROUND: Kidney has long been thought to be a body's largest organ of elimination for maintaining acid-base balance. In recent years, the research on kidneys has mainly focused on the structural characteristics of the kidney of single age group animals. In this paper we used histological and immunohistochemical methods to observe and compare the structure characteristics of yak kidney and the expression of epidermal growth factor receptor (EGFR), bone morphogenetic protein-2 (BMP-2) and p53 in the kidney of yaks of three different age groups. The aim of the study was to investigate histological characteristics of age-related chan- ges in the kidney of yak and expression and localisation of kidney-related factors. MATERIALS AND METHODS: Fifteen healthy male and female yaks from highland plateaus (three groups: newborn, adult and old yaks, n = 5 per group). Histo- logical methods were used to compare the relevant characteristics of the kidney of yaks. The immunohistochemistry method was used to observe the expression and localisation of EGFR, BMP-2, and p53 of the kidney of different ages, and the optical density value was measured and analysed by using image analysis software. RESULTS: This is an overall observation of the kidney tissue section, which includes the surface of the renal capsule and the internal parenchyma. In the renal parenchyma, there are renal corpuscles, renal tubules. The internal substance included cortex and medulla, which were bounded by the arched artery. In the cortex, there were renal corpuscles, convoluted part of renal tubules (proximal convoluted tubule and distal convoluted tubule) and collecting tubules. The medulla included straight parts of renal tubules (proximal straight tubule and distal straight tubule), thin segments and collecting tubules. It was observed that the organisational structure of the kidney of yaks did not change with age, but the degree of development of the internal structure (glomeruli, renal tubules and collecting tubules) of the kidney changed with age. Immunohistochemical results demonstrated that EGFR and BMP-2-positive reaction in the newborn group was mainly distributed in the proximal tubule epithelial cells, and widely distributed in the adult and old groups. However, the p53-positive reaction was widely distributed in the newborn, adult and old groups. CONCLUSIONS: The results revealed that the kidney structure tended to be com- pleted with age, and the function of the kidney gradually improved. EGFR and BMP-2 had the effect of promoting kidney development. However, p53 had been widely distributed in the newborn kidney of the yaks. It is suggested that p53 had been involved in cell migration and metabolic differentiation and self-renewal in the new stage.

Effect of Modified Si Junzitang Drug Serum on Expression of Apoptosis-related Molecules of Gastric Cancer Cell SGC-7901 / 中国实验方剂学杂志

Objective: To explore the effect of modified Si Junzitang (MSJZT) drug serum on the expression of apoptosis-related molecules of gastric cancer cell SGC-7901 and further its anti-tumor mechanism.Method: A total of 40 SD rats were randomly divided into four groups:low-dose,middle-dose,high-dose MSJZT (0.213,0.426,0.853 g·kg-1) groups and normal group (n=10).The treatment groups were administrated through gastric perfusion,and the normal group was given the equivalent volume of normal saline for 10 days.1.5 h after the last treatment,chloral hydrate peritoneal anesthesia was performed,blood was collected from heart,and different doses of serum were separated to prepare drug-containing serum of low-dose,middle-dose,high-dose MSJZT groups,in order to incubate SGC-7901 gastric cancer cell.Early and late apoptosis rates were detected with flow cytometry.Afterwards,the tumor suppressor gene p53,c-nucleoprotein gene (c-Myc),cysteine-aspartic acid protease-3(Caspase-3),B-cell lymphoma-2(Bcl-2) mRNA expressions were confirmed by fluorescence quantitative polymerase chain reaction (Real-time PCR).The protein expressions of p53,c-Myc,Caspase-3,Bcl-2 were detected by immunofluorescence.Result: Compared with the normal group,the high-dose MSJZT group could obviously increase the apoptosis rate to 22.58%(PPPPPPConclusion: MSJZT drug serum could exert an anti-tumor effect by inhibiting the expression of the anti-apoptotic protein Bcl-2,and promoting the expressions of pro-apoptotic-related molecules p53,c-Myc,Caspase-3.

The challenge for allogeneic hematopoietic stem cell transplantation in acute leukemia/ myelodyplastic syndrom with TP53, TET2 or DNMT3A gene mutations / 天津医药

@#Recently, much gene mutations have been detected in patients with acute leukemia or myelodysplastic syndrome (MDS) using next-generation sequencing (NSG) technology. Some of them are proved to be important prognostic markers. It has been showed that TP53, TET2 or DNMT3A gene mutations are associated with poor prognosis in acute leukemia or MDS patients. The prognosis of these patients is poor with short remission and survival. Allogeneic hematopoietic stem cell transplantation is the only way to cure these patients. However, the outcomes after transplantation are inferior to those in patients without these mutations. The hypomethylating agents or immune targeting therapy might improve their prognosis when combined with the present strategies. Here, the impact of TP53, TET2 and DNMT3A gene mutations on the prognosis after chemotherapy or transplantation is reviewed.

Dual-cyclical nucleic acid strand-displacement polymerization based signal amplification system for highly sensitive determination of p53 gene.

In the present study, we proposed a novel dual-cyclical nucleic acid strand-displacement polymerization (dual-CNDP) based signal amplification system for highly sensitive determination of tumor suppressor genes. The system primarily consisted of a signaling hairpin probe (SHP), a label-free hairpin probe (LHP) and an initiating primer (IP). The presence of target DNA was able to induce one CNDP through continuous process of ligation, polymerization and nicking, leading to extensively accumulation of two nicked triggers (NT1 and NT2). Intriguingly, the NT1 could directly hybridize SHP, while the NT2 could act as the target analog to induce another CNDP. The resulting dual-CNDP contributed the striking signal amplification, and only a very weak blank noise existed since the ligation template of target was not involved. In this case, the target could be detected in a wide linear range (5 orders of magnitude), and a low detection limit (78 fM) was obtained, which is superior to most of the existing fluorescent methods. Moreover, the dual-CNDP sensing system provided a high selectivity towards target DNA against mismatched target and was successfully applied to analysis of target gene extracted from cancer cells or in human serum-contained samples, indicating its great potential for practical applications.

Mutations in and Expression of the Tumor Suppressor Gene p53 in Egg-Type Chickens Infected With Subgroup J Avian Leukosis Virus.

To investigate the molecular mechanisms of the oncogenic effects of avian leukosis virus subgroup J (ALV-J), we examined mutations in and the expression of p53 in the myelocytomas distributed in the liver, spleen, trachea, and bone marrow, as well as in fibrosarcomas in the abdominal cavity and hemangiomas in skin from chickens that were naturally or experimentally infected with ALV-J. Two types of mutations in the p53 gene were detected in myelocytomas of both the experimentally infected and the naturally infected chickens and included point mutations and deletions. Two of the point mutations have not been reported previously. Partial complementary DNA clones with a 122-bp deletion in the p53 gene ORF and a 15-bp deletion in the C-terminus were identified in the myelocytomas. In addition, moderate expression of the mutant p53 protein was detected in the myelocytomas that were distributed in the liver, trachea, spleen, and bone marrow. Mutant p53 protein was not detected in the subcutaneous hemangiomas or in the abdominal fibrosarcomas associated with natural and experimental ALV-J infection, respectively. These results identify mutations associated with abnormal expression of p53 in ALV-J-associated myelocytomas, suggesting a role in tumorigenesis.

Effect of SIRT1 on cell proliferation and senescence of human pancreaticβ-cells CRL-1837 / 中华内分泌代谢杂志

Histone deacetylase 1 ( sirtuin 1, SIRT1) is an important member of deacetylase family, and plays an important role in the process of malignant tumor and embryonic development. In this article it was found that overexpression of SIRT1 could accelerate the DNA synthesis in human pancreatic beta cell CRL-1837 and inhibit cell senescence. SIRT1 also could bind to p53 as detected by co-immunoprecipitation and could change the phosphorylation level of p53.

Study on gallic acid induced human hepatoma SMMC-7721 cells apoptosis and its mechanism / 中国药理学通报

Aim To investigate the proliferative effect and the apoptosis of human hepatoma SMMC-7721 cells induced by gallic acid ( GA ) , and its underlying mechanism. Methods SMMC-7721 cells were cul-tured in vitro. MTT assay was used to observe the pro-liferation of SMMC-7721 cells induced on GA 24 , 48 , 72 h. The morphological and ultra structural changes of the SMMC-7721 cells were observed by inverted micro-scope and transmission electron microscope respective-ly. Annexin V-FITC/PI staining was used to quantify the percentages of apoptosis in the total cell popula-tion. The expression of p53 mRNA was investigated by RT-PCR. Western blot was used to determine the pro-tein expression of p53. Results GA(6. 25~50 μmol ·L-1 ) markedly inhibited the activity of proliferation and induced apoptosis of SMMC-7721 cells after 48 h in a dose-dependent manner. GA significantly induced cell nuclear condensation and fragmentation. RT-PCR and Western blot results showed that GA could improve the expression of p53 mRNA and protein. Conclusion GA can inhibit the proliferation of human hepatoma SMMC-7721 cells and induce cells apoptosis. The mechanism may be associated with improving tumor suppressor gene p53 expression.

Selenium dioxide inducing apoptosis of cervical cancer cells by regulating apoptosis-related proteins / 重庆医学

Objective To investigate the inducing effects of selenium dioxide(SeO2 ) on the apoptosis in human cervical carcino-ma cell line Hela and its influence on the expression of apoptosis-related proteins caspase-3 and P53 .Methods Hela cells were trea-ted with different concentrations of SeO2 for 24 h in vitro ;the morphological changes of Hela cells were observed by the optical mi-croscope;the influence of SeO2 on the cell proliferation and vitality was examined by the MTT assay ;the flow cytometry was em-ployed to detect the cell apoptosis rate ;the expressions of caspase-3 and P53 proteins in Hela cells were determined by the Western blot analysis .Results Under the optical microscopy ,SeO2 generated the obvious influence on the cell growth morphology ,a large number of cells became rounded and shrunken ,and lost the normal form ,while the adherence cell number was evidently decreased and the proliferation was slowed down ;the MTT results showed that SeO2 markedly inhibited the cell proliferation and viability in a dose-dependent manner ,in which ,the cell apoptosis rates induced by the 0 ,1 .875 ,3 .750 ,7 .500 ,15 .000 and 30 .000 μmol/L con-centrations of SeO2 were 3 .12% ,30 .56% ,33 .42% ,37 .50% ,45 .43% and 69 .38% respectively ,which revealing the obviously in-creasing trend;the Western blot assay revealed that SeO2 could up-regulate the caspase-3 and P53 levels ,and reached the peak value at the concentration of 7 .500μmol/L .Conclusion SeO2 could induce the cervical cancer cell apoptosis possibly by up-regulating the expressions of caspase-3 and p53 in Hela cells .

Imunoexpressão das proteínas COX-2, p53 e caspase-3 em adenoma colorretal e mucosa não neoplásica / Immunoexpression of the COX-2, p53, and caspase-3 proteins in colorectal adenoma and non-neoplastic mucosa

OBJETIVO: Analisar a imunoexpressão das proteínas COX-2, p53 e caspase-3 em adenomas colorretais e na mucosa não neoplásica. MÉTODOS: Foram submetidos à colonoscopia 72 indivíduos que forneceram 50 amostras de adenomas e 45 de mucosa colorretal não neoplásica. Os tecidos foram obtidos pela técnica de arranjo em matriz (tissue microarray) e submetidos a estudo imunoistoquímico com anticorpos primários p53, COX-2 e caspase-3. A positividade e intensidade da imunorreação foram classificadas. Foram estudadas as seguintes variáveis: localização do adenoma no colo, grau de displasia, tamanho, e escores de positividade e intensidade da imunoexpressão das proteínas p-53, caspase-3 e COX-2. RESULTADOS: Nos adenomas, a imunoexpressão da proteína p53 mutada foi positiva em 30 (60%) e negativa em 20 (40%) amostras. Na mucosa colorretal não neoplásica, a imunoexpressão da proteína p53 mutada foi negativa em 39 (86,6%) amostras e positiva em 6 (13,3%) (p<0,0001). Houve diferença significativa entre o maior tamanho (p=0,006) e o maior grau de displasia dos adenomas (p<0,0001) e a intensidade de imunoexpressão da proteína p53 mutada. A positividade e intensidade da imunoexpressão das proteínas COX-2 (p=0,14) e caspase-3 (p=0,23), nos adenomas e na mucosa colorretal não neoplásica, não apresentaram diferença significante. CONCLUSÃO: A proteína p53 mutada é hiperexpressada nos adenomas em comparação com a mucosa não neoplásica. Nos adenomas, o maior tamanho e o maior grau de displasia foram associados à maior expressão da proteína p53 mutada. A imunoexpressão das proteínas COX-2 e caspase nos adenomas não apresentou correlação com os aspectos anatomopatológicos e não foi diferente em termos de níveis de expressão correspondentes na mucosa não neoplásica.

OBJECTIVE: To analyze the immunoexpression of the COX-2, p53, and caspase-3 proteins in colorectal adenomas and non-neoplastic mucosa. METHODS: 72 individuals were subjected to colonoscopy, which provided 50 samples of adenomas and 45 samples of non-neoplastic colorectal mucosa. The tissue samples were obtained via the tissue microarray technique and subjected to immunohistochemical analysis using primary anti-p53, anti-COX-2, and anti-caspase-3 antibodies. The positivity and intensity of the immunoreaction were classified. The analyzed variables were as follows: site of the adenomas in the colon, degree of dysplasia, size, and score of positivity and intensity of immunoexpression of the p-53, caspase-3, and COX-2 proteins. RESULTS: The immunoexpression of mutated protein p53 was positive in 30 (60%) adenoma samples and negative in 20 (40%) adenoma samples. The immunoexpression of mutated protein p53 was negative in 39 (86.6%) samples and positive in 6 (13.3%) samples of the non-neoplastic colorectal mucosa (p<0.0001). Significant differences were seen between both the largest size (p=0.006) and the highest degree of dysplasia (p<0.0001) of the adenomas and the intensity of immunoexpression of mutated protein p53. The positivity and intensity of immunoexpression of COX-2 (p=0.14) and caspase-3 (p=0.23) showed no significant differences between the adenomas and the non-neoplastic colorectal mucosa. CONCLUSION: Mutated protein p53 was hyperexpressed in the adenomas compared with the non-neoplastic mucosa. Greater size and greater degree of dysplasia in the adenomas were associated with higher expression of mutated protein p53. The immunoexpression of COX-2 and caspase-3 in the adenomas did not exhibit a correlation with the anatomical-pathological features of the tumors and did not differ from the corresponding expression levels in the non-neoplastic mucosa.

Investigação do gene p53 de frangos expostos às aflatoxinas / Investigation of p53 gene in poultries exposed to aflatoxins

Identificou-se o efeito das aflatoxinas (AFs) sobre o gene p53 de frangos de corte, de linhagem comercial, separados em: grupo experimental, tratado (GT) com ração comercial contendo 2,8ppm de AFs totais durante 21 dias consecutivos, e grupo-controle (GC), sem exposição às AFs. Macroscopicamente, as alterações caracterizaram-se por hepatomegalia e aspecto pálido-amarelado com alguns focos hemorrágicos e, histologicamente, por desarranjo trabecular, pleomorfismo hepatocítico com cariomegalia, degeneração vacuolar intracitoplasmática, necrose com infiltração linfocítica e hiperplasia de ductos biliares. A PCR com os primers GSPT53c-1 com base no gene candidato a p53 (GenBank XM_424937.2) gerou um produto de aproximadamente 350 pares de base. O amplicon sequenciado a partir do DNA dos frangos do GT não apresentou mutação ou deleção, assim como padrão de bandas do PCR-RFLP não foi distinto entre ambos os grupos experimentais e a sequência depositada no banco de genes. Os resultados sugerem que não ocorreu transversão devido à exposição às AFs no fragmento amplificado. Conclui-se que a PCR-RFLP e o sequenciamento do produto da PCR não são ferramentas apropriadas para diagnóstico da exposição de frangos às AFs nas condições experimentais empregadas.

To identify the effects of aflatoxins (AFs), Cobb lineage poultry were separated in an experimental group in which they were treated with commercial ration containing 2.8ppm of total AFs during 21 days (TG) and a control group without AFs exposure (CG). In the liver of poultries exposed to AFs, alterations were microscopically observed, which were characterized by hepatomegaly, a pale yellowish aspect with some hemorrhagic spots, and histologically a trabecullar disarranging pleomorphic hepatocytes with cariomegaly, intracytoplasmatic vacuolar degeneration, necrosis, lymphocytic infiltration and hyperplasia of biliary ducts. The PCR with GSPT53c-1 primers based on p53 candidate gen (GenBank XM_424937.2) generated a product of approximately 350 base pairs. The sequenced amplicon obtained from the DNA of treated poultry did not display any mutation or deletion, and the PCR- RFLP bands patterns were also not distinct in both experimental groups. The results indicated that transversion did not occur in the fragment amplified due to AFs exposure. As a consequence of results obtained with p53 gene (NM_205264.1) we concluded that PCR-RFLP and sequencing of PCR product are not appropriate diagnostic tools for the detection of poultry exposure to AFs, at least in the experimental conditions performed.

The role of p53 in induction of growth arrest DNA damage-inducible gene 45β in human hepatoma cells by oxaliplatin / 中华肝胆外科杂志

Objective To identify the role of p53 in the induction of growth arrest DNA damage-inducible gene 45β (GADD45β) in HCC cells by Oxaliplatin.Methods A Hep3B+p53 clone was established by transfection of the full-length p53 sequence to Hep3B.Following oxaliplatin administration,quantitative real-time PCR was employed to validate the expression changes of GADD45β.pGL3 basic luciferase plasmids including promoter fragments were synthesized in vitro and transfected into cells.The effects on promoter activity,cell growth and the cleavage of Caspase-3 were further focused on.Results Hep3B+p53 expressed p53 protein stably.The transfection of p553 enhanced the induction of GADD45β in Hep3B by Oxaliplatin.The promoter activity of fragments constructed NF-κB and E2F-1 binding sites was induced about 1.5 and 0.8 folds by transfection of p53.The colony formation and DNA syntheses were inhibited apparently in Hep3B+p53 with p53 by Oxaliplatin (30.41％ and 75.60％ by 100 μmol/L Oxaliplatin,respectively).Moreover,p53 transfection triggered cleavage of Caspase-3 more rapidly.Conclusion p53 played a role in the induction of GADD45β in Hep3B by Oxaliplatin.

Expression and significance of NDRG1, p53 and VEGF in esophageal squamous cell carcinoma / 肿瘤研究与临床

ObjectiveTo study the expression and significance of NDRG1(N-myc downstream regulated gene-1), p53 and VEGF in esophageal squamous cell carcinoma (ESCC). MethodsNDRG1, p53 and VEGF protein were detected by immunohistochemistry (IHC, SP method) in 20 cases of normal esophageal squamous epithelium and 78 cases of ESCC.ResultsThe results of IHC shows that in normal esophageal squamous epithelium and esophageal squamous cell carcinoma,the positive rate of NDRG1 was 100.0 ％(20/20) and 55.1％ (43/78) respectively, p53 was 0 (0/20) and 65.4 ％ (51/78) respectively, VEGF was 30.0 ％(6/20)and 67.9 ％(53/78)respectively,all had statistical significance.There was inverse correlationof NDRG1 expression and lymphatic invasion(r =-0.237,P = 0.036).However expression of NDRG1 was no statistical significance with patient' s age,gender,grade,TNM stage,patient' s five year survival.The expression of p53 was inverse correlated with NDRG1,and the expression of VEGF was inverse correlated with NDRG1 (r =-0.331, P = 0.003). ConclusionNDRG1 may be a new tumor suppress gene and play an important role in the development and metastasis of ESCC.

Expression and clinical significance of EZH2 and p53 protein in human prostate cancer / 肿瘤研究与临床

ObjectiveTo explore the expression of EZH2 and p53 protein in primary prostate cancer (Pca) and its clinical significance.Methods High-throughput tissue microarray technique and immunohistochemistry was used to detect the expression of EZH2 and p53 protein in 48 human prostate cancer specimens without a history of chemo-radiation therapy and 15 cases of benign prostate hyperplasic (BPH) tissues. The pathological characteristics and the relationship of the expression of EZH2 and p53 protein in primary prostate cancer was analyzed. ResultsImmunohistochemical results showed that the positive rates of EZH2 and p53 protein in prostate cancer were 87.50 ％ (42/48) and 33.33 ％ (16/48), respectively, which were significantly higher than that in BPH tissues[13.33 ％ (2/15) and 0 (0/15)](x2=26.429, x2=5.058,P ＜0.05). The expression of EZH2 and p53 protein was significantly related to Gleason score, TNM stage (P ＜0.05), but not to age and serum prostate-specific antigen (PSA) level (P ＞0.05). The positive expression in patients with Gleason＞6 was higher than that with Gleason≤6(P ＜0.05).The positive expression in patients with T3-T4 stage was higher than that with T1-T2 stage(P ＜0.05).Spearman rank correlation showed a significantly positive correlation between EZH2 and p53 protein (r=0.294, P ＜0.05). ConclusionEZH2 and p53 protein may participate in the pathogenesis of prostate cancer.The overexpression of EZH2 and p53 protein could become an index for the evaluation of the level of malignancy and progression of prostate cancer.Furthermore,combining detection of EZH2 and p53 protein may provide a new theoretical basis for the treatment of prostate cancer.