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Expression of hMSH2 gene and mutant p53 in sporadic digestive tract tumors

Chinese Medical Journal 2003年第1期116卷 53-56页

作者：康燕婕张振科王俊霞陈静彭勃康萍 Department of Gastroenterology Hebei Medical University

Objective To investigate the role of mutated mismatch repair gene hMSH2 and mutant p53 gene in the carcinogenesis and development of sporadic digestive tract tumors. Methods hMSH2 gene in normal and tumor tissue of 30 digestive tract tumor specimens was examined using polymerase chain reaction-single strand conformation polymorphism (pCR-SSCp) silver staining. The pCR product with an abnormal strand was sequenced directly. mutant p53 protein in the tumor tissue was analyzed immunohistochemically. Results Six patients were identified as having mutated strands, three on hMSH2 exon 1 and three on hMSH2 exon 5. DNA sequencing revealed that all 6 patients had mutated basic groups that led to decrease in function of the hMSH2 protein. Forty percent (12/30) of patients were p53 positive. The frequency of mutated hMSH2 in p53 positive patients (41.7%) was significantly higher than in p53 negative patients (5.6%, pplays an important role in the carcinogenesis and development of digestive tract tumors through stimulating p53 mutation.

关键词： digestive tract tumor mismatch repair gene hMSH2 mutant p53 gene

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Experimental research for specific down-regulated expression of p53 gene by individual antisense RNA in vitro

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The Chinese-German Journal of Clinical Oncology 2007年第1期6卷 62-67页

作者： YahongWang Shaofeng Xu Yuanyuan Zhang Bin Zhang Yumei Feng Ruifang Niu Li Fu Department of pathology and Research Laboratory of Breast Cancer Key Laboratory of Breast Cancer Prevention and Therapy of the Ministry of Education Tianjin Medical University Tianjin 300060 China

Objective： To investigate the specific blockage effect of individual antisense RNA on mutant p53 gene in vitro. Methods： The single strand antisense transcription system containing mt-p53 exon 8 sequence （pGEM3zf（＋/-）p53exon8） was constructed. The ligation of antisense RNAwith mt-p53 gene was confirmed by in situ hybridization; MDA-MB-231 human breast cancer cells were transfected with ASp53exon8＇RNA cotionic liposome-mediated. Expression of mt-p53 protein was examined by immunocytochemical staining and Western blot. Cell proliferation was evaluated by MTT assay; Cell cycle distribution was determined by flow cytometry （FCM）; Apoptosis was observed by TUNEL. Results： In transfected MDA-MB-231 cells, hybridization signals were observed in cytoplasm. ASp53exon8＇RNA transfection induced inhibition of cell proliferation, G2/M phase arrest and increasing apoptotic rates. In addition, expression of p53 protein was down-regulated. Conclusion： pGEM3zf（＋/-）p53exon8 was well constructed and ASp53exon8＇RNA can block mt-p53 gene expression specifically and then inhibit MDA-MB-231 cell proliferation in vitro, which may serve as therapeutic means for human malignancy.

关键词： individual antisense RNA mutant p53 gene specific blockage mutant protein expression

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