Detection of germline mutations of hMLH1 and hMSH2 based on cDNA sequencing in China.

AIM To detect the germline mutations of hMLH1 and hMSH2 based on mRNA sequencing to identify hereditary non-polyposis colorectal cancer (HNPCC) families. METHODS Total RNA was extracted from peripheral blood of 14 members from 12 different families fulfilling Amsterdam criteria II. mRNA of hMLH1 and hMSH2 was reversed with special primers and heat-resistant reverse transcriptase. cDNA was amplified with expand long template PCR and cDNA sequencing analysis was followed. RESULT Seven germline mutations were found in 6 families (6/12, 50%), in 4 hMLH1 and 3 hMSH2 mutations (4/12, 33.3%); (3/12, 25%). The mutation types involved 4 missense, 1 silent and 1 frame shift mutations as well as 1 mutation in the non-coding area. Four out of the seven mutations have not been reported previously. The 4 hMLH1 mutations were distributed in exons 8, 12, 16, and 19. The 3 hMSH2 mutations were distributed in exons 1 and 2. Six out of the 7 mutations were pathological, which were distributed in 5 HNPCC families. CONCLUSION Germline mutations of hMLH1 and hMSH2 can be found based on cDNA sequencing so as to identify HNPCC family, which is highly sensitive and has the advantages of cost and time saving.

[1]  Weimin Sun,et al.  High-throughput gene sequencing assay development for hereditary nonpolyposis colon cancer. , 2004, Clinical colorectal cancer.

[2]  P. Peltomäki,et al.  Mutations Associated with HNPCC Predisposition — Update of ICG-HNPCC/INSiGHT Mutation Database , 2004, Disease markers.

[3]  M. Genuardi,et al.  Hereditary nonpolyposis colorectal cancer and related conditions , 2003, American journal of medical genetics. Part A.

[4]  M. Loda,et al.  Evaluation of Microsatellite Instability and Updated Version Cited Articles Citing Articles E-mail Alerts Evaluation of Microsatellite Instability and Immunohistochemistry for the Prediction of Germ-line Msh2 and Mlh1 Mutations in Hereditary Nonpolyposis Colon Cancer Families , 2022 .

[5]  W. Farrell,et al.  Hereditary non-polyposis colorectal cancer: an updated review. , 2000, European journal of surgical oncology : the journal of the European Society of Surgical Oncology and the British Association of Surgical Oncology.

[6]  J. G. Park,et al.  Risk of gastric cancer in hereditary nonpolyposis colorectal cancer in Korea. , 2000, Clinical cancer research : an official journal of the American Association for Cancer Research.

[7]  A. de la Chapelle,et al.  Genetic susceptibility to non-polyposis colorectal cancer , 1999, Journal of medical genetics.

[8]  P. Møller,et al.  Familial endometrial cancer in female carriers of MSH6 germline mutations , 1999, Nature Genetics.

[9]  L. Aaltonen,et al.  Cancer risk in mutation carriers of DNA‐mismatch‐repair genes , 1999, International journal of cancer.

[10]  T. Smyrk,et al.  Colorectal carcinoma survival among hereditary nonpolyposis colorectal carcinoma family members , 1998, Cancer.

[11]  M. Nöthen,et al.  A Novel Missense Mutation in the DNA Mismatch Repair Gene hMLH1 Present among East Asians but Not among Europeans , 1998, Human Heredity.

[12]  P. Peltomäki,et al.  Mutations predisposing to hereditary nonpolyposis colorectal cancer: database and results of a collaborative study. The International Collaborative Group on Hereditary Nonpolyposis Colorectal Cancer. , 1997, Gastroenterology.

[13]  P. Peltomäki,et al.  Genotype and phenotype in hereditary nonpolyposis colon cancer: a study of families with different vs. shared predisposing mutations , 2004, Familial Cancer.

[14]  Jeremy R. Jass,et al.  HNPCC and Sporadic MSI-H Colorectal Cancer: A Review of the Morphological Similarities and Differences , 2004, Familial Cancer.

[15]  H. Olsson,et al.  Defective mismatch-repair in patients with multiple primary tumours including colorectal cancer. , 2003, European journal of cancer.

[16]  K. Shin,et al.  Mutational analysis of promoters of mismatch repair genes hMSH2 and hMLH1 in hereditary nonpolyposis colorectal cancer and early onset colorectal cancer patients: identification of three novel germ-line mutations in promoter of the hMSH2 gene. , 2002, Cancer research.

[17]  J. Kładny,et al.  Optimization of experimental conditions for RNA‐based sequencing of MLH1 and MSH2 genes , 2001, Human mutation.

[18]  H. Lynch,et al.  GENETIC SUSCEPTIBILITY TO NONPOLYPOSIS COLORECTAL CANCER , 1999 .