The Resistance Research And Expression Of C×43 In Bone Marrow Of Patients With Acute Leukemia

BackgroudIn recent years, through the conventional chemotherapy and bone marrow transplanttreatment, the prognosis of leukemia has changed greatly, but because the generation of drug-resistant, it is difficult for a part of patients to relieve or relapse after remission soon after.Leukemia cell is the proliferation of malignant cloned hematopoietic cells, its survival,differentiation, proliferation and bone marrow stromal cells are closely related. Cx43 is the majorconnection protein of bone marrow stromal cells. a close relation has been found betweenreduction or elimination Cx43 and the occurrence, development and transfer of multiple tumors.Studies have shown that gap junction intercellular communication (GJIC) mediated by Cx43plays an important role of regulation to leukemia cell proliferation, apoptosis and drug sensitivity.the specific mechanism of Cx43 in tumor drug resistance is not very clear, remains to be furtherresearch [2, 3]. The generation of drug resistance is many factors and many steps, many genesthat work together, a large number of studies have identified P-gp and COX-2 are overexpress-tion and Participate in generation of drug-resistant.in a variety of malignant tumor. Our study isUsing SYBR Green real-time quantitative reverse transcription polymerase Chain reaction(SYBR-RT-PCR), the expression of Cx43、P-gp and Cox-2mRNA were detected in 77 ALpatients at different phases, Including 37 initially-treated, to further detect the relationship ofthe occurrence, development and resistance of leukemia. Through the mechanism study of bonemarrow stromal cells and tumor cells resistance offer a possible means for refractory bloodcancer diagnosis and treatment.MethodsAccording to the experiment diagnostic criteria ZhangZhinan: initially treated , complete- remission group, the relapsed group, the normal group 4 groups, testing the following related inde-x:1．Seperated and cultured the normal and、leukemic、complete-remission、relapsed humanBMSCs in virto,evaluated the expression of Cx43 mRNA and protein by PCR.2. Real-time quantitative fluorescence PCR (Real-time PCR) directly test the expression of Cx-43、P-gp、COX-2 mRNA of the normal and leukemic、complete-remission、relapsed group.Result1. Real time-polymerase chain reaction (PCR) detect Cx43mRNA of the experimental sampl-es stromal cell:cx43mRNA Expression levels of initially-Treated and relapse AL Patients were lower than those in normal patients and CR patient(p<0.01).2. Real time-PCR directly detect Cx43, P-gp, COX-2 mRNA expression of bone marrow ,The results show: Expression levels of cx43mRNA in initially-treated and relapse AL Patientswere lower than those in normal patients and CR patients, with statistical significance (initially–treated were0.001、0.004；relapse Patients were P<0.001),while the comparison between normalpatients and CR patients showed no statistical significance (P=0.185). Expression levels of P-gp、COX-2mRNA in initially-treated AL Patients were higher than those in normal patients, withstatistical significance ( p=0.035、0.019)；Expression levels of P-gp、COX-2mRNA in relapsePatients were higher than those in normal patients(P<0.001) ).3. Cx43mRNA Expression level was negatively correlated with P-gp and Cox-2mRNA, r=-0.544、r=-0.406, P<0.001.4. A follow-up of four months was conducted in 36 initially-treated patients, eight of themdied. The expression cx43 in those who died was lower than that who survived. the differencebeing significant(t=2.16, P=0.042).Conclusion1. This experiment tested the different course CX43 expression from cell culture and clinicalbone marrow, the results is consistent trend in future for the clinical research provides a simpleand convenient to experiment the method.2. CX43mRNA Expression levels of initially-Treated and relapse AL Patients were lowerthan those in normal patients and CR patient, P-gp and Cox-2mRNA Expression levels of initial-ly-Treated and relapse AL Patients were higher than those in normal patients and CR patient.CX43 expression probably is a favorable prognostic factor.Cx43 is participation in the genesis,development and drug resistance of AL.3. the interaction between Cx43 and tumor cells regulate expression of P-gp and COX-2 ,which affect the chemotherapy drug reaction to the tumor cells.