The Methylation Level Of Survivin Gene, PDCD5Gene And NOR1Gene In Multiple Myeloma

Background and Objective:Multiple myeloma (MM) as one species of malignant hyperplastic diseases that characterized by the proliferation of monoclonal plasma cells in bone marrow, at present the pathogenesis ulteriorly has not yet been explicated. In recent years, the study found that there is a close relationship between the occurrence and development with methylation in multiple myeloma (MM). In order to explore its significance, we intends to test the methylation level of survivin gene, PDCD5gene and NOR1gene in multiple myeloma patients, control group and KM3cell lines.Methods:There were5cases bone marrow samples of multiple myeloma patients which collected from the Third Hospital of Central South University(are all referral patients) from August2012to December2012, Among these Samples,3cases are male and2cases are female, the age from46to70years old and median age is58years.The control group are made up of5cases of bone marrow samples of patients with non-hematologic malignancies,1cases are male and4cases are female, the age from22to58years old and median age is40years, In these patients,the number of patients with iron deficiency anemia is4,the number of patients with megaloblastic anemia is1. We used the KM3cell line as a specimen for cell culture.Used the exponential phase cells to extract genomic DNA.Adopting the saturated phenol chloroform method to extract genomic DNA of the bone marrow specimens of MM patients, the bone marrow samples of patients with non-hematologic malignancies and KM3cells.Adopting BSP method to detect the methylation level of survivin gene, PDCD5gene and NOR1gene in MM patients, control group and KM3cell lines.Results:According to the methylation site index value=mCpG/+mCpG (CPG), We obtained the methylation pattern of CpG site of CPG island of survivin gene, PDCD5gene and NOR1gene in upstream promoter regions through BSP method, the results are as follows:the results are as follows:1) The methylation levels of the CpG island of survivin promoter region in bone marrow samples of5cases multiple myeloma patients were1.0%,0.5%,0.0%,0.5%,0.0%, The methylation levels of the CpG island of survivin promoter region in bone marrow samples of5cases patients with non-hematologic malignancies were0.5%,1.5%,2.5%,1.0%,2.5%, the methylation level of the CpG island of survivin promoter region in KM3cell line was0.0%.2) The methylation levels of the CpG island2in PDCD5promoter region of positive-sense strand promoter in bone marrow samples of5case multiple myeloma patients were0.0%,1.8%,0.0%,1.4%,0.0%, The methylation levels of the CpG island2in PDCD5promoter region of positive-sense strand promoter in bone marrow samples of5cases patients with non-hematologic malignancies were0.0%,0.0%,0.0%,1.4%,0.0%, the methylation level of the CpG island2in PDCD5promoter region of positive-sense strand promoter in KM3cell line was0.0%.3) The methylation levels of the CpG island3in NOR1promoter region of positive-sense strand promoter in bone marrow samples of5cases multiple myeloma patients were0.0%,3.8%,3.8%,0.0%,2.5%, The methylation levels of the CpG island3in NOR1promoter region of positive-sense strand promoter in bone marrow samples of5cases patients with non-hematologic malignancies were0.0%,1.2%,0.0%,0.0%,0.0%, the methylation level of the CpG island3in NOR1promoter region of antisense strand promoter in KM3cell line was100%.Conclusions:1) The CpG island of survivin promoter region shows non-methylation or low methylation in MM patients and patients with non-hematologic malignancies, and shows non-methylation in KM3cell lines.2) The CpG island2in PDCD5promoter region of positive-sense strand promoter shows non-methylation or low methylation in MM patients and patients with non-hematologic malignancies, and shows non-methylation in KM3cell lines.3) The CpG island3in NOR1promoter region of positive-sense strand promoter shows non-methylation or low methylation in MM patients and patients with non-hematologic malignancies, but the CpG islands3in NOR1promoter region of antisense strand promoter shows high methylation in KM3cell lines.