Expression And Clinical Significance Of CD137L In Multiple Myeloma Cells

Part I Expression and clinical significance of CD137 L in multiple myeloma cells【Objective】(1) To determine expression and clinical significance of CD137 L molecular in patients with monoclonal gammopathy of undetermined significance(MGUS) and multiple myeloma cells;(2)To explore function of CD137 L in multiple myeloma cell lines.【Methods】(1) The expression of CD137 L molecule on myeloma cells/normal plasma cell surface was detected by flow cytometry;(2) Clinical significance of CD137 L molecule expressed by multiple myeloma cells was accessed via rank sum test; expression level of high/low of CD137 L on overall survival was evaluated through survival analysis and Log-Rank test;(3) SiRNA, the customization of SiRNA for CD137 L gene, transfected myeloma cell lines U266, RPMI-8226, KMS-11 by Lipo3000.Then the expression of CD137 L was detected by RT-PCR; cell cycle distribution after inhibition of CD137 L signal was detected by PI; cell proliferation was detected by CCK8.【Results】(1) Fresh bone marrow specimens of 28 patients with newly diagnosed multiple myeloma patients were collected. The expression of CD137 L molecule on CD45-/CD38+/CD138+ cell group in bone marrow was detected, and the median expression level was 29(7-94)%; the expression of CD137 L molecule on 9 patients with MGUS was 7(2-57)%;(2) That the expression level of CD137 L between patients with MGUS and newly diagnosed MM showed statistic difference indicated that it could be as a marker for differential diagnosis; the different expression level by rank sum test between those with newly diagnosed MM and post-treated MM, post-treated MM and RRMM indicated that it could be a marker for MRD;(3) The follow-up of patients found that after the treatment CD137 L level of 11/13 patients decreased, and these patients at least achieved PR;(4) there is no related with the level of CD137 L and type, ISS stage, DS stage, white blood cell count, hemoglobin concentration, platelet count, serum beta 2- microglobulin, lactate dehydrogenase, serum albumin, calcium concentration, the ratio of bone marrow plasma cell by correlation analysis;(5) According to median values of CD137 L expression level, all the newly diagnosed patients were divided into two groups, low level expression and high level and survival analysis showed no significant difference by Log-Rank test. The 2 years survival rate of low level group and the high one was 84.7%, 74.1%;(6) KMS-11, RPMI 8226,U266 cells were transfected using Lipo3000 and only U266 cell line was inhibited obviously. Inhibition of CD137 L induced cell proliferation by CCK8 test and a distribution change of G1 and S phase on cell cycle.【Conclusions】Multiple myeloma cell lines and primary myeloma cells had a high expression level of CD137 L, MGUS cells had a low level while normal plasma cells surface without CD137 L expression; CD137 L can be a marker for diagnosis of MM and MGUS and for minimal residual disease; CD137 L expression of MM patients had no correlation with clinical and biological features; In vitro the inhibition of CD137 L signaling on U266 cells can induce cells proliferation.Part II Efficacy and prognosis of the treatment with PAD vs. PDD in patients with the newly diagnosed multiple myeloma【Objective】To explore the therapeutic effect and prognosis of PAD and PDD regimen as induction therapy in 116 patients with newly diagnosed multiple myeloma.【Methods】116 patients of newly diagnosed multiple myeloma receiving PAD and PDD scheme(bortezomib + Adriamycin or liposomal doxorubicin + dexamethasone) were analysed to compare curative effect and prognosis of the two groups for induction therapy,【Results】Overall response of PAD and PDD regimen as induction therapy in the newly diagnosed multiple myeloma patients was 95.5%、93.5% respectively. Survival analysis showed that there was no statistical difference between the two groups of PFS and OS curves. But the median PFS and OS in PAD group were 46 and 63 months respectively, and the median PFS and OS in PDD group were not reached. Following up to 58 months, the cumulative survival rate of PAD group was 60.88%, while the PDD group reached 89.12%. Comparation of PAD and PDD for patients with high-risk via FISH test or sequential autologous hematopoietic stem cell transplantation showed no difference. The median PFS of patients with ISS stage III in the PDD group was not reached, while in the PAD group it was only 35.5 months.【Conclusions】Patients with newly diagnosed multiple myeloma treated with PAD and PDD regimen as induction therapy showed no difference with efficacy and prognosis.Part III Infections on induction therapy in Hospitalised Patients with Multiple Myeloma: Main Characteristics and Risk Factors【Objective】To explore characteristic and risk factors of multiple myeloma patients on the induction therapy.【Methods】Clinical data of 116 patients with symptomatic MM treated by PDD or PAD regimen in our hospital from June 2008 to June 2015 were collected and the clinical characteristics and risk factors were explored.【Results】The incidence of infection was 83.6% and the mortality rate was 3.1%. With the development of treatment, rate of infection showed a downward trend. The incidence of site of infection from high to low in turn is respiratory system, skin and mucosa, oral and digestive tract, septicemia, urinary tract; the proportion of gram positive bacteria and gram negative bacteria is considerable; and infection was due to bacterial translocation of the normal flora. Incidence of herpes zoster was 13.8%. The rate of infection during patients with subcutaneous injection about bortezomib was significantly lower than intravenous way; patients with PDD scheme have a higher infection rate than those with PAD scheme.【Conclusions】During induction period, the incidence of infection is very high, and infection is the major cause of mortality. The main infection sites are lung, clinicians and patients need to strengthen the prevention and treatment of respiratory infection.