Expression Of Autophagy - Related Genes In Chronic Lymphocytic Leukemia And The Effect

Objective:To explore the clinic significance of BECN1 and ATG5 mRNA in the patients with chronic lymphocytic leukemia (CLL).Methods:One hundred patients with CLL and fifty healthy controls were enrolled in the present study. The mRNA levels of BECN1 and ATG5 were detected by realtime polymerase chain reaction (RT-PCR) with SYBR Green. The p53 deletion and other cytogenetic abnormalities were detected using florescence in situ hybridization (FISH). We analysed p53 mutation and immunoglobulin variable heavy chain (IGHV) mutation status using PCR and direct sequencing. Flow cytometry was used to detect the expression of CD38 and ZAP-70.Results:Of the 100 CLL patients and 50 controls, the median expression of BECN1 mRNA was 0.108 and 0.035, respectively, and the median expression level of ATG5 mRNA was 0.084 and 0.066, respectively. The CLL samples expressed higher levels of BECN1 and ATG5 mRNA compared with healthy controls. BECN1 expression level was associated with Binet stage (P=0.047). ATG5 expression level was closely associated with Binet stage (P=0.001), ZAP-70 (P=0.008), CD38 (P=0.007), p53 aberrations (P=0.014). There was difference between the lower and higher ATG5 mRNA expression level (P=0.020) in time to first treatment (TTT). In Cox multivariate analysis, Binet stage (P=0.001) was the only independent prognostic factor for TTT.Conclusion:Expression levels of autophagy-associated genes BECN1 and ATG5 mRNA were up-regulated in CLL, which suggests higher autophagy level in CLL cells. The high expression of ATG5 gene probably plays an important role in the prognosis of CLL.Objective:This study was aimed to explore the effects and its mechanisms of chidamide on the proliferation, autophagy and apoptosis of chronic lymphocytic leukemia (CLL) cells.Methods:CLL cells from the peripheral blood of CLL patients were sorted by Ficoll. The CLL cells were cultured in vitro with chidamide. The cell proliferation was detected by CCK-8 method; the protein levels of LC3 in cells were analyzed by Western blot; the cell apoptosis was analyzed by Annexin V/PI double staining and multiparameter flow cytometry. Quantitative RT-PCR was used to evaluate the mRNA changes of BECN1 and ATG5.Results:The findings indicated that chidamide inhibited the viability of CLL cells in time- and concentration-dependent manners. The median apoptotic rates of the negative control (NC),3-MA, chloroquine (CQ), chidamide, chidamide and CQ treatment groups were 20.97%,29.83%,25.33%,37.47%,44.37%, respectively. Western blot analysis showed that chidamide resulted in the decline of autophagy in CLL cells by detecting LC3. The median BECN1 mRNA expression of the NC、 3-MA, CQ, chidamide, chidamide and CQ treatment groups were 0.078,0.062, 0.054,0.051,0.055, respectively. The median ATG5 mRNA expression of the NC, 3-MA, CQ, chidamide, chidamide and CQ treatment groups were 0.087,0.034, 0.056,0.078,0.107, respectively.Conclusion:These results showed that chidamide is a potent antitumor agent for treating CLL. Chidamide could induce cell apoptosis and decline autophagy during the chidamide -induced death of CLL cells.