Naturally derived sulforaphene inhibits proliferation of mantle cell lymphoma by down-regulating CRM1 expression / 肿瘤

ABSTRACT

Objective: To investigate the expression level of chromosomal region maintenance 1 (CRM1) in mantle cell lymphoma, and to explore the possible mechanism of naturally derived sulforaphene LFS-01 inhibiting the proliferation of mantle cell lymphoma cells. Methods: The expression of CRM1 in mantle cell lymphoma was analyzed by Oncomine data mining. After treatment with LFS-01, the viability of mantle cell lymphoma JeKo-1 cells was detected by CCK-8 assay, the nuclear transport function of CRM1 protein was measured by laser confocal microscopy, the expression of CRM1 protein was detected by Western blotting, the cell cycle arrest and apoptosis were analyzed by FCM and transmission electron microscopy, the expression change of apoptosis pathway-related proteins was detected by Western blotting, and the impact of caspase inhibitor z-VAD-FAM on the effects of LFS-01 was detected by CCK-8 assay and FCM. Lentiviral infection was used to establish a stable JeKo-1 cells expressing mutant CRM1 (C528S), then the effects of LFS-01 on the nuclear transport function of CRM1 and the proliferation of JeKo-1 cells were detected by laser confocal microscopy and CCK-8, respectively. The transcriptional level in JeKo-1 cells after LFS-01 treatment was detected by RNAseq, and the viability of JeKo-1 cells treated with Toll-like receptor (TLR) inhibitor TAK-242 and LFS-01 was measured by CCK-8 assay. Results: CRM1 was overexpressed in mantle cell lymphoma (P < 0.05). LFS-01 inhibited the proliferation of JeKo-1 cells, and the 50% inhibitory concentration (IC50) at 24 h and 48 h were 5.81 μmol/L and 9.09 μmol/L, respectively. 20.0 μmol/L LFS-01 inhibited the nuclear transport function of CRM1 (P < 0.05), 6.0 μmol/L LFS-01 down-regulated the expression level of CRM1 (P < 0.05), 10.0 μmol/L LFS-01 induced cell cycle arrest at G2/M phase and induced apoptosis (both P < 0.05), and 4 μmol/L LFS-01 up-regulated the expression levels of poly ADP-ribose polymerase (PARP), cleaved caspase-3 and cleaved caspase-9 (all P < 0.01). The caspase inhibitor z-VAD-FAM reversed the apoptosis-induction effect of LFS-01 (P < 0.01). CRM 1 mutation eliminated the effects of LFS-01 on CRM1 function and cell proliferation (both P < 0.05). LFS-01 inhibited the cell proliferation-related pathways (P < 0.01), and TLR inhibitor TAK-242 combined with LFS-01 had synergetic inhibitory effect on JeKo-1 cells (P < 0.01). Conclusion: LFS-01 can inhibit the growth of mantle cell lymphoma cells through inducing cell cycle arrest and apoptosis, and CRM1 is essential in this process. In addition, LFS-01 and TLR inhibitor TAK-242 have synergetic inhibitory effect on mantle cell lymphoma cells.