TRIP13基因mRNA在慢性淋巴细胞白血病B淋巴细胞的表达及其调控JVM-2细胞增殖和凋亡的分子机制探讨

OBJECTIVE: To investigate the clinical significance of expression level of thyroid hormone receptor interactors 13 (TRIP13) gene to probe its function and downstream molecular mechanism in chronic lymphocytic leukemia (CLL). METHODS: Real-time quantitative PCR method was used to detect the expression levels of TRIP13 mRNA of CD19(+) B lymphocytes in 30 cases of patients with CLL and 12 cases of peripheral blood hematopoietic stem cell donors (normal control group). Lentivirus mediated shRNA was used to interference the mRNA and TRIP13 protein in CLL cells JVM-2. Scramble sequence was used as control. Methyl thiazolyl tetrazolium colorimetric assay (MTT) and flow cytometry was used to detect the cell proliferation and apoptosis in TRIP13 knocked-down and negative control JVM-2 cells. RESULTS: TRIP13 mRNA level was significantly higher in 30 cases of CLL patients (2(−ΔCt)=0.014 89) compared with 12 healthy donors (2(−ΔCt)=0.000 19) (P<0.001). Validated TRIP13 shRNA target was achieved in JVM2 cell. Compared with the control group, down-regulation of TRIP13 expression could significantly inhibit the proliferation of JVM-2 cells and induce apoptosis. The expressions of Myc and Bcl-2 protein in JVM-2 cells decreased significantly after interference with TRIP13 (P<0.001), and the expressions of Bax, caspase 3 and Bad protein increased significantly (P<0.001). CONCLUSION: TRIP13 mRNA significantly over-expressed in CLL patients CD19(+) B lymphocytes. TRIP13 could influence JVM2 cell proliferation and apoptosis through proliferation- and apoptosis-related proteins.