DEPTOR-mTOR信号通路介导的自噬在多发性骨髓瘤中对破骨细胞分化的调控作用

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Abstract Objective To study the role of DEPTOR knockdown in RPMI-8226 cells on the protein expression of RANKL and differentiation of THP-1 into osteoclast-like cells in a contactless co-culture system and its possible mechanism. Methods Constructed DEPTOR shRNA expression vector GV115-shRNA was transferred into RPMI-8226 cell to produce packaged lentivirus. Western blot was applied to measure the protein levels of DEPTOR and RANKL. The expression of autophagy-associated proteins LC-3 and Atg5 were confirmed by Western blot analysis. Three groups were divided：THP-1 group, THP-1 + RPMI-8226 group and THP-1 + DEPTOR shRNA group. Osteoclast-like cells were identified by TRAP. The mRNA levels of calcitonin receptor (CTR) and Cathepsin-K were examined using RT-PCR. Results The results showed that protein expression levels of DEPTOR and RANKL were significantly lower in RPMI-8226 cells transfected with GV115 DEPTOR shRNA compared with that in untransfected cells (P < 0.05). The expression levels of autophagy-associated proteins LC-3 and Atg5 in the DEPTOR shRNA group were significantly lower than those in the control shRNA group and the parental group (P < 0.05). In the co-culture system, THP-1 cell could differentiate into TRAP positive multinuclear cells. RPMI-8226 promoted mRNA expression of CTR and Cathepsin-K (P < 0.05). DEPTOR shRNA suppressed osteoclast-like cells formation and decreased CTR and Cathepsin-K mRNA expression in co-cultures, the differences were statistically significant (P < 0.05). Conclusion In the coculture system, DEPTOR shRNA inhibits the differentiation of THP-1 cells into TRAP positive multinuclear cells, which may be due to its inhibition on RANKL expression in RPMI-8226 cells, and the inhibition of autophagy will restrain osteoclast maturation.

Key words： Myeloma bone disease DEPTOR RNA interference Autophagy THP-1

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	ZHANG Haoran1 QIAO Xuxu1 BI Minghong1 ZHAI Yunzhi1 QIAN Liyu2 PAN Chengwu2 ZHAO Lun1

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ZHANG Haoran1 QIAO Xuxu1 BI Minghong1 ZHAI Yunzhi1 QIAN Liyu2 PAN Chengwu2 ZHAO Lun1. The role of DEPTOR-mTOR signaling pathway-mediated autophagy on osteoclasts differentiation and maturation in multiple myeloma[J]. 中国医药导报, 2018, 15(4): 18-22.

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http://www.yiyaodaobao.com.cn/EN/ OR http://www.yiyaodaobao.com.cn/EN/Y2018/V15/I4/18

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