Brief Reports
Copyright ©2005 Baishideng Publishing Group Co., Limited. All rights reserved.
World J Gastroenterol. Jan 14, 2005; 11(2): 275-279
Published online Jan 14, 2005. doi: 10.3748/wjg.v11.i2.275
Polymerase chain reaction-single strand conformational polymorphism analysis of rearranged during transfection proto-oncogene in Chinese familial hirschsprung’s disease
Tao Guan, Ji-Cheng Li, Min-Ju Li, Jin-Fa Tou
Tao Guan, Ji-Cheng Li, Department of Lymphology, Institute of Cell Biology, Zhejiang University Medical College, Hangzhou 310031, Zhejiang Province, China
Min-Ju Li, Jin-Fa Tou, Children抯 Hospital, Zhejiang University Medical College, Hangzhou 310006, Zhejiang Province, China
Author contributions: All authors contributed equally to the work.
Supported by the Fund for Excellent Young Talented Persons by Public Health Ministry of China, and Analysis and Testing Foundation of Zhejiang Province, No. 99075
Correspondence to: Professor. Ji-Cheng Li, Department of Lymphology, Institute of Cell Biology, Zhejiang University Medical College, Hangzhou 310031, Zhejiang Province, China. lijc@mail.hz.zj.cn
Telephone: +86-571-87217451 Fax: +86-571-87217145
Received: March 15, 2004
Revised: March 18, 2004
Accepted: May 13, 2004
Published online: January 14, 2005
Abstract

AIM: To investigate the relationship between mutations of rearranged during transfection (RET) proto-oncogene and Chinese patients with Hirschsprung’s disease (HD), and to elucidate the genetic mechanism of familial HD patient at the molecular level.

METHODS: Genomic DNA was extracted from venous blood of probands and their relatives in two genealogies. Polymerase chain reaction (PCR) products, which were amplified using specific primers (RET, exons 11, 13, 15 and 17), were electrophoresed to analyze the single-strand conformational polymorphism (SSCP) patterns. The positive amplified products were sequenced. Forty-eight sporadic HD patients and 30 normal children were screened for mutations of RET proto-oncogene simultaneously.

RESULTS: Three cases with HD in one family were found to have a G heterozygous insertion at nucleotide 18974 in exon 13 of RET cDNA (18974insG), which resulted in a frameshift mutation. In another family, a heterozygosity for T to G transition at nucleotide 18888 in the same exon which resulted in a synonymous mutation of Leu at codon 745 was detected in the proband and his father. Eight RET mutations were confirmed in 48 sporadic HD patients.

CONCLUSION: Mutations of RET proto-oncogene may play an important role in the pathogenesis of Chinese patients with HD. Detection of mutated RET proto-oncogene carriers may be used for genetic counseling of potential risk for HD in the affected families.

Keywords: Hirschsprung’s disease; Proto-oncogene proteins RET; Transfection; PCR-SSCP