[The analysis for mRNA mutation of MLH1, MSH2 genes and the gene diagnosis for hereditary nonpolyposis colorectal cancer].

Abstract

OBJECTIVE:

To identify hereditary nonpolyposis colorectal cancer (HNPCC) families based on the germline mutations of MLH1 and MSH2 mRNA.

METHODS:

RNA was extracted from the peripheral blood of the 14 members from 12 different families fulfilling Amsterdam Criteria II. The germline mutations of MLH1 and MSH2 mRNA were detected by cDNA sequencing analysis following reverse transcription-PCR(RT-PCR) with special primers, heat-resistance reverse transcriptase, and expand long template PCR. DNA was extracted from the peripheral blood of the 14 members, the corresponding exons, in which mutations were found using the above method, were amplified with Taq enzyme, sequencing analysis was followed.

RESULTS:

Six germline mutations were detected and identified from the 6 different families based on mRNA, 4 of them to be in MLH1, the other 2 in MSH2. The MLH1 mutations distribute in the exon 8, 12, 16, and 19. The MSH2 mutations distribute in exons 1 and 2. The 6 mutations were identified from the corresponding exons respectively in genomic DNA sequencing analysis. The mutation types involve in 4 missense, 1 silent, and 1 non-coding area mutations. Five out of the 6 mutations have not been reported previously. Five out of the 6 mutations were pathological, involving in 5 different families. The five families were identified to HNPCC families.

CONCLUSION:

HNPCC family can be identified with RNA-based sequencing of MLH1 and MSH2 from peripheral blood, which has the advantages of both cost, time saving and high sensitivity.

• Authors: Chao-fu Wang, Xiao-yan Zhou, Tai-ming Zhang, Meng-hong Sun, Ye Xu, Da-ren Shi
• Journal: Zhonghua yi xue yi chuan xue za zhi = Zhonghua yixue yichuanxue zazhi = Chinese journal of medical genetics (Zhonghua Yi Xue Yi Chuan Xue Za Zhi) Vol. 23 Issue 1 Pg. 32-6 (Feb 2006) ISSN: 1003-9406 [Print] China
• PMID: 16456782 (Publication Type: Journal Article)

Chemical References

• Adaptor Proteins, Signal Transducing
• Biomarkers, Tumor
• Carrier Proteins
• MLH1 protein, human
• Neoplasm Proteins
• Nuclear Proteins
• RNA, Messenger
• MSH2 protein, human
• MutL Protein Homolog 1
• MutS Homolog 2 Protein

Topics

• Adaptor Proteins, Signal Transducing
• Biomarkers, Tumor (genetics)
• Carrier Proteins (genetics)
• Colorectal Neoplasms, Hereditary Nonpolyposis (diagnosis, genetics)
• Female
• Germ-Line Mutation
• Humans
• Male
• MutL Protein Homolog 1
• MutS Homolog 2 Protein (genetics)
• Mutation
• Neoplasm Proteins (genetics)
• Nuclear Proteins (genetics)
• RNA, Messenger (analysis)