Study On Aberrant Methylation Of P15Gene In Multiple Myeloma

Objective:The experiment is objected to explore the pathogenesis of Multiple myeloma(MM), especially the tumor Suppressor gene(p15gene)CpG islands methylation on occurrence and development of MM and The signifieance or evaluating prognosis and directing therapy.Meth()ds:1.It was Collected from January2009to June2011confirmed54cases of newly diagnosed.referral of MM bone marrow samples of patients,diagnostic criteria. refer to the Zhang Zhi nan edited by "blood disease diagnosis and the efficacy of standard",typing stage grouping(all patients from the seeond and the First Affiliated Hospital of the Kunming Meedical Colleege).Also set up the control group were nutritional anemia and newly diagnoses idiopathic thrombocytopenic purpura in patients with bone marrow specimens. Methylation-specific polymerase chain reaction (MSP) and bisulfite modification seequeencing technology studies in patientsi with MM bone marrow of p15gene promoter region methyfation status of the use of automated sequencing technology to its base methylation analysis of amplifiedl produets weere sequeenced the results SPSS17.0package for statistical analysis.2. To design and synthesize specific primer of m (methylation system).and u (nonmethylation system)for real time PCR reaction,and further use of real-timet florescence quantitative PCR to monitor thee mRNA expression,the correlations of results were analyxed by SPSS17.0package. Result:1. The results showed that patients with MM p15gene promoter region methylation status:15of54(27.78%) myeloma samples showed p15hypermethylation in the5'promoter region. None of positive result (0.00%.0/40) is detected in the control group, there is significant difference between the two group(P<0.05);2. Methylation-positive specimens were modified direct sequencing test result:there are19sites methylated in the5' promoter region in p15gene.3.We found the promoter methylation status of p15gene in MM had no Obviously relationship with the newly-diagnosed, the recurrence, the gender,the age. the stage was not obvious, according to the above clinical and pathological features of grouping within each group showed no significant difference(each group P>0.05).4.RT-qPCR results:for relative expression of p15mRNA, median±quartile of MM patients samples is28.64±28.77Fold, the control group is25.79±78.16Fold: methylated group is28.64±19.13Fold, and unmethylated group is29.86±29.24Fold: newly-diagnosed sample is23.92±26.46Fold, treated sample is36.13±27.65Fold; Unfortunately, all of three group data have no correlations(P>0.05).Conclusion:1. This study of54cases of multiple myeloma patients, there is the p15gene promoter region methylation. statistically significant compared with control group:its overall positive rate of methylation with gender, age. stage, group has nothing to do, study suggestsp15abnormal methylation may play a role in MM pathogenesis.2. In the test, there are no correlation of expression of p15mRNA between MM patients and the control group. Moreover, between methylated MM group and unmethylated group, or new-diagnosed patients and treated patients are still no significantly differences. Thus transcription of p15might be regulated by other molecular mechanism.