The Study Of Promoter Of Human LCRG1 Gene Cloning ,Mutation And Methylation In Laryngeal Squamous Cell Carcinoma

LCRG1(laryngeal carcinoma related gene1,LCRG1),a new candidate tumor suppressor gene of Laryngeal Carcinoma. we analyzed the promoter function at the following aspects:1 to analyze the crucial sequence which participate in the transcriptional regulation by use of 5'-deletion mutagenesis;2 to measure the DNA mutation of the promoter CpG island in Laryngeal Carcinomas by use of PCR-SSCP (single strand conformation polymorphism, SSCP); 3 to measure the DNA methylation status of the promoter CpG island in Laryngeal Carcinomas by use of methylation specific PCR (MSP); to measure the expression status in Laryngeal Carcinomas by use of semi-quantitive reverse transcription PCR (RT-PCR).In the study of the LCRG1 gene, we have shown that:1, the minimal promoter of LCRG1 gene was defined to a fragment from-169 to -57 position relative to the transcription start site;2, No mutation was found in the promoter sequence of the LCRG1 gene in laryngeal carcinoma.3, methylation of LCRG1 gene promoter was found in 5.00% Laryngeal Carcinoma.4, LCRG1 gene was significantly down-regulated in 16 of 40 (40%) primary laryngeal carcinomas.Conclusion:1 the minimal promoter of LCRG1 gene was defined to a fragment from-169 to -57 position relative to the transcription start site;2 No mutation was found in the promoter sequence of the LCRG1 gene in laryngeal carcinoma ,which indicate the mutation of the promoter sequences of the LCRG1 gene may not contribute to abnormal expression of LCRG1 gene in laryngeal carcinoma;3 methylation of LCRG1 gene promoter was found in 5.00% Laryngeal Carcinoma , methylation of LCRG1 gene promoter may be involved in the regulatory mechanisms of LCRG1 gene expression.