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protective effects of proanthocyanidins on beta-amyloid peptide (25-35)-induced pC12 cell apoptosis by blocking S-phase and increasing p53 gene expression

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Neural Regeneration Research 2010年第2期5卷 108-112页

作者： Hanfang Mei Zhaoyang Xie Qifeng Zhu Department of Biochemistry and Molecular Biology Guangdong Pharmaceutical University Guangzhou 510006 Guangdong Province China School of pubfic Health and Tropical Medicine Southem Medical University Guangzhou 510515 Guangdong Province China Institute of Biochemistry and Molecular Biology Guangdong Medical College Zhanjiang 524023 Guangdong Province China

BACKGROUND： Current studies related to the effects of proanthocyanidins on Alzheimer＇s disease have focused primarily on the signal transduction pathway of cellular apoptosis. However, the influence of p53 gene expression on cell cycle regulation, with regard to the protective mechanisms of proanthocyanidins, has not been reported. OBJECTIVE： To observe the effect of proanthocyanidins on cell cycle distribution, cellular apoptosis and p53 gene expression in β-amyloid peptide （25-35）（Aβ25-35）-induced pC12 cells cultured in serum-free media, and to investigate the molecular neuroprotective mechanisms of proanthocyanidins with regard to cell cycle regulation. DESIGN, TIME AND SETTING： A parallel, controlled, at the Institute of Biochemistry and Molecular Biology cellular, and molecular study was performed Guangdong Medical College from July 2006 to July 2008. MATERIALS： proanthocyanidins were provided by Nanjing Xuezi Medical and Chemical Research Center, China; Aβ25-35 was provided by Sigma, USA; pC12 cells were provided by the Institute of Basic Medical Science, Academy of Military Medical Sciences; and rabbit anti-p53 polyclonal antibody was provided by Santa Cruz Biotechnology, USA. METHODS： pC12 cells were cultured in serum-free media for 24 hours. Cells from the model group were treated with 25 μmol/L Aβ25-35 for 24 hours. Cells in the drug protection group were pre-treated with 30 mg/L proanthocyanidins for 1 hour and then treated with 25 μmol/LAβ2^-35 for 24 hours. The control group was not treated. MAIN OUTCOME MEASURES： Flow cytometry was used to detect cell cycle distribution and rate of apoptosis; reverse-transcriptase polymerase chain reaction was used to detect p53 mRNA expression; and Western blot was used to detect p53 protein expression. RESULTS： After treating with 25 μmol/LAβ25-35 for 24 hours, the rate of apoptosis and the percentage of cells in S phase were significantly increased （p 〈 0.01 ）, and p53 mRNA and protein expressions were decreased. p

关键词： proanthocyanidins β-amyloid peptide (25-35) Alzheimer's disease pC12 cells p53 gene neural regeneration

Successful management of postoperative recurrence of hepatocellular carcinoma with p53 gene therapy combining transcatheter arterial chemoembolization

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World Journal of Gastroenterology 2005年第24期11卷 3803-3805页

作者： Yong-SongGuan YuanLiu LongSun XiaoLi QingHe DepartmentofRadiology WestChinaHospitalSichuanUniversityChengdu610041SichuangProvinceChina

Transcatheter arterial chemoembolization (TACE) has become the standard treatment for unresectable hepatocellular carcinoma (HCC). But this method has some shortages. p53 gene, which was found to be mutant in many human tumors, has been proved with broadspectrum anti-tumor effects. We reported a 23-year-old patient with recurrent HCC after irregular hepatectomy. The p53 gene was applied to this patient. We injected percutaneously and infused transcatheterally p53 gene (Gendicine, Shenzhen Sibiono Bentech, China) into his recurrent nodules in liver respectively and 4 d later, the patient received TACE therapy. In the 2 mo follow-up, the patient was in good clinical condition with normal liver function and no recurrence was identified. The case report proposed that recurrent HCC could be successfully treated with p53 gene therapy combining TACE.

关键词： Hepatocellular carcinoma Transcatheter arterial chemoembolization p53 gene

Correlation of p53 gene mutation and expression of p53 protein in cholangiocarcinoma

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World Journal of Gastroenterology 2006年第29期12卷 4706-4709页

作者： Xiao-Fang Liu Hao Zhang Shi-Guang Zhu Xian-Ting Zhou Hai-Long Su Zheng Xu Shao-Jun Li Department of Hepatobiliary Surgery The Affiliated Yantai Yuhuangding Hospital of Qingdao University Medical College Yantai 264000 Shangdong Province China Department of general Surgery The First Affiliated Hospital of China Medical University Shenyang 110001Liaoning Province China

AIM： To characterize the tumor suppressor gene p53 mutations and study the correlation of p53 gene mutation and the expression of p53 protein in cholangiocarcinoma. METHODS： A total of 36 unselected, frozen samples of cholangiocarcinoma were collected, p53 gene status（exon 5-8） and p53 protein were examined by automated sequencing and immunohistochemical staining, combined with the clinical parameters of patients. RESULTS： p53 gene mutations were found in 22 of 36 （61.1%） patients. Nineteen of 36 （52.8%） patients were positive for p53 protein expression. There were significant differences in extent of differentiation and invasion between the positive and negative expression of p53 protein. However, there were no significant differences in pathologic parameters between the mutations and non-mutations. CONCLUSION： The alterations of the p53 gene evaluated by DNA sequence analysis is relatively accurate. Expression of p53 protein could not act as an independent index to estimate the prognosis of cholangiocarcinoma.

关键词： Cholangiocarcinoma p53 gene Mutation DNA sequence

Expression of Retroviral Mediated Wild Type p53 gene in Laryngocarcinoma and Fibrosarcoma Cell Lines and Its Inhibitory Effects on Cancer Growth

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High Technology Letters 1996年第1期2卷 103-106页

作者：陈晓巍 Dept.of Otolaryngology Peking Union Medical College HospitalBeijing 100730P.R.China

The accumulated evidence indicates that head and neck cancers arise due to multiple genetic changes in both dominant oncogenes and tumor suppressor genes. In this report we examined whether the wild-type p53 gene was able to suppress in vitro and in vivo cellular growth of laryngocarcinoma cell line and fibrosarcoma which carried multiple genetic abnormalities. Introduction of retroviral mediated wild-type p53 complementary DNA into laryngocarcinoma cell line (Hep2) and fibrosarcoma cell line (MDAH 041) carrying either a homozygous deletion or a missense in the p53 gene greatly suppressed tumor cell growth and the malignant phenotype of the cancer cells was reversed.

关键词： p53 gene Laryngocarcinoma Fibrosacoma

DETECTION OF p53 gene MUTATION IN pLASMA OF pATIENTS WITH GASTRIC CANCER

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Chinese Journal of Cancer Research 2004年第3期16卷 182-187页

作者：苏鹏程李子禹张连海万文徽任晖张桂国王怡邓国仁季加孚 Department of Surgical Oncology Peking University School of Oncology Beijing Institute for Cancer Research Beijing Cancer Hospital Department of Clinical Laboratory Peking University School of Oncology Beijing Institute for Cancer Research Beijing Cancer Hospital Dapartment of Biochemistry Peking University School of Oncology Beijing Institute for Cancer Research Beijing Cancer Hospital

Objective: To investigated p53 gene mutation in plasma of gastric cancer patients. Methods: DNA extracted from plasma and matched tumor and tumor-adjacent non-cancerous tissues of 96 gastric cancer patients, and DNA from 20 healthy volunteers were studied. Exon 5, 6, 7, and 8 of p53 were amplified by polymerase Chain Reaction (pCR). The mutation status was analyzed by denaturing high-performance liquid chromatography (DHpLC), followed by direct sequencing of cases with aberrant chromatographic patterns. Results: Heterozygous mutations of p53 gene were detected in 19.9% (19/96) of primary tumor tissues and 5.2% (5/96) of corresponding plasma. All p53 gene mutations detected in plasma DNA consisted with mutations in the matched primary tumor samples. Neither the tumor-adjacent gastric mucosa tissues nor control plasma from healthy volunteers showed p53 gene mutation. No correlation was found between p53 mutation status and clinicopathological features of gastric cancer patients. Conclusion: p53 gene mutation in plasma can be detected in tissues and plasma of gastric cancer patients, which could be applied in screening and surveillance of this disease.

关键词： p53 gene Mutation, plasma DNA Gastric cancer

THE　SIGNIFICANCE　OF　p53　gene　MUTATIONS　AND　EXpRESSIONS　IN　HUMAN　COLORECTAL　TUMORS

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Chinese Journal of Cancer Research 1996年第2期8卷 109-112页

作者：钱桦郁宝铭周锡庚王瑞年黄薇陈赛娟陈竺 Department of Surgery Xinhua Hospital Shanghai Second Medical University Shanghai Department of Surgery Ruijin Hospital SSMU Shanghai Department of pathology SSMU Shanghai Department of Hematology Ruijin Hospital SSMU Shanghai

Using a polymerase chain reaction-single strand conformation polymorphism (pCR-SSCp) approach we analyzed 18 human colorectal adenocarcinomas for mutations in exons 5,6,7,8 of p53 gene. At the same time,p53 gene product expression was studied immunohistochemically in these 18 case in frozen sections. The expression of p53 protein was also immunohistochemically studied in formalin-fixed paraffin embeded spccimens of 76 colorectal adenocarcinomas and 112colorectal polyps. Eigbt out of 18 cases (44%) showed a variant band indicative of a mutation in exons 5-6 of p53gene 7 out of 8 cascs (88%) with p53 gene mutations were positivelystanined for p53. There was no significant correlation between p53. expression and clinicopathological manifestations and prognosis. but the strongest staining was cncountered in those cases with well differentiated and early stage adenocarcinomas,while weaker staining was encountered in poorly differentiated and mucoid adenotarcinomas. p53expression was not observed in proliferative polyps and adenomas with low grade dysplasia. The frequency of p53expression reached 88% (ppes of adenomas, p53 expression was most frequent in villous type (pp53 expression in adenoma, adenoma with malignant change and adenocarcinoma were 4%, 88% and 51% *** indicate that genetic changes of p53 gene play an important role in the transformation from benign adeuoma to adenocarcinoma. p53 immunohistochcmistry can be used as a surrogate marker for p53 gene mutation for early discovery of colorectal adenocarcinomas.

关键词： Colorectal neoplasms p53 gene Mutation,Expression pCR-SSCp

EXpRESSION OF p53 gene IN ORAL SQUAMOUS CELL CARCINOMA AND ITS RELATION WITH CLINICAL AND pATHOLOGICAL pARAMETERS AND pROGNOSIS OF pATIENTS

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Chinese Medical Sciences Journal 1995年第4期10卷 199-203页

作者：毛驰卢勇赖钦声夏雨和杨橙 pUMCHospital CAMS＆PUMCBeijing100730 WestChinaUniversityofMedicalSciences ChengduSichuan610044

One hundred and eleven cases of oral squamous cell carcinoma (OSCC) were examined for overexpression of p53 protein by using immunohistochemical technique. Association between p53 protein overexpression and clinical and pathological parameters as well as prognosis of patients were also analyzed. p53 protein overexpression was commonly observed (69. 4%) in OSCC and may be used as a marker of carcinogenesis of OSCC. The level of p53 protein overexpression is correlated with the lower three and five-year survival rate of OSCC. The presence or absence of p53 overexpression was not correlated with sex, age, site of tumor, size of tumor, degree of differentiation,node status,and clinical stage in OSCC. Single factor COX proportional hazards regression model analysis indicated that there was no significant association between p53 overexpression and prognosis of OSCC. Multivariable COX model analysis failed to establish effective life function or risk rate function. These showed that all the parameters analyzed in this study as well as p53 overexpression were not significant and effective risk factors of prognosis for patients with OSCC.

关键词： oral squamous cell carcinoma p53 gene

RELATIONSHIp　BETWEEN　THE　MUTATION　OF　p53　gene　AND　INFILTRATION，METASTASIS　AND　pROGNOSIS　OF　GASTRIC　CARCINOMA

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Chinese Journal of Cancer Research 1996年第4期8卷 263-266页

作者：辛彦赵凤凯吴东瑛王艳萍徐蕾张荫昌 Cancer Institute China Medical University Shenyang Department of Histopathology Charring Cross & Westminster Medical School University of London UK

gastric carcinomas were examined immunohistochemically with McAb to p53 protein in order to investigate the relationship between the expression of p53 protein and histological differentiation of gastric carcinoma, and to approach the mechanism of infiltration and metastasis of gastric carcinoma. The results showed that nuclear expression of p53 protein was significantly related to tumor size, depth of invasion, lymph node and liver metastases; but not related to histological differentiation. It is suggested that the accumulation of p53 protein was increased with the progression of gastric carcinoma, and therefore the cancer clone with p53 gene mutation may play an important role in the development of tumor invasion and metastasis.

关键词： p53 gene Tumor suppresser gene.Metastasis Gastric carcinoma

GROWTH INHIBITION OF HUMAN LARYNGEAL CANCER CELL WITH THE ADENOVIRUS-MEDIATED p53 gene

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Chinese Journal of Cancer Research 1999年第3期11卷 157-160页

作者：王琪韩德民王文革吴祖泽张伟 Department of Otolaryngology-Head and Neck Surgery Institute of Otolaryngology Beijing Tongren Hospital Capital University of Medical Sciences Beijing China

Objective: In most laryngeal cancers, the function of p53 gene is down regulated. To explore the potential use of p53 in gene therapy of laryngeal cancer, by introducing wild-type p53 into laryngeal cancer cell line via a recombinant adenoviral vector, Ad5CMV-p53 and analyzing its effects on cell and tumor growth. Methods: A human laryngeal cancer, cell line Hep-2 was used. Recombinant cytomegalovirus-promoted adenoviruses containing human wild-type p53 cDNA was transiently introduced into Hep-2 line. The growth suppression of the Hep-2 cells and established s.c. squamous, carcinoma model was examined. The p53 protein expression was detected using immunohistochemical analysis. Results: The transduction efficiencies of Hep-2 cell line were 100% at a multiplicity of 100 or greater. The p53 protein expression peaked on day 2 after infection and lasted far 5 days. In vitro growth assays revealed cell death following Ad5CMV-p53 infected. In vivo studies, Ad5CMV-p53 inhibited the tumorigenicity of Hep-2 cell, and in nude mice with established s.c. squamous, carcinoma, nodules showed that tumor volumes were significantly reduced in mice that received peritumoral infiltration of Ad5CMV-p53. Conclusion: Adenovirus-mediated antitumor therapy carrying the p53 gene is an efficient method to inhibit laryngeal cancer growth. Transfection of laryngeal cancer cells with the wild-type p53 gene via Ad5CMV-p53 is a potential novel approach to the therapy of laryngeal cancer.

关键词： gene therapy Laryngeal cancer p53 gene Adenoviruses