| 摘要 |
The present invention utilizes human gene sequence, a specific CpG island of RBL2 retinoblastoma-like 2 (pl30) gene, and designs primers based on base mismatch principle selected to constitute CpG island methylation test reagent kit for diagnosing tumor and screening demethylation medicine. The reagent kit includes primers specific for MSP- PCR reaction system. The complete CpG island sensitive to methylation is also provide. Moreover specific primers for detection of "hot-spot" mutations at exon 1, identified as tumor biomarkers and the PCR reaction are provided. Three RBL2 retinoblastoma-like 2 (pl30) regions rich in CpG dinucleotide, the promoter region immediately 5'- flanking to the transcription start site (ATG) , the exon 1 and the intron 1, are target to epigenetic events in tumors. Two specific exon 1 mutations, inside a CpG region sensitive to methylation, are present in different cancer hystotype representing tumor-specific marker. Analysis of methylation patterns in the RBL2 retinoblastoma-like 2 (pl30) regions rich in CpG dinucleotide by Methylation Specific PCR (MSP) and mutational analysis of exon 1 provide methods for diagnosis and prognosis and for targeting therapy in cancer patients. |
