MiR-148a participates in the growth of RPMI8226 multiple myeloma cells by regulating CDKN1B
MiR-148a participates in the growth of RPMI8226 multiple myeloma cells by regulating CDKN1B
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DOI：
10.1016/j.biopha.2016.11.002
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发表时间：
2016-12-01
影响因子：
7.5
通讯作者：
Nie, Yuling
中科院分区：
文献类型：
--
作者：
Lang, Tao;Nie, Yuling
目的：为探讨miR-148 a对多发性骨髓瘤（MM）细胞系RPMI 8226细胞增殖和细胞周期的影响及其分子机制，采用RT-PCR和Western blotting方法检测miR-148 a和CDKN 1B在MM细胞和正常骨髓原代细胞中的表达。采用流式细胞仪检测miR-148 a基因敲减MM细胞和正常MM细胞的增殖和细胞周期。在正常和miR-148 a敲低的MM细胞中测定p-NPAT、p-Rb和p-CDC 6的蛋白表达。结果：MM细胞中miR-148 a的表达水平明显高于正常骨髓原代细胞，而CDKN 1B的表达水平明显低于正常骨髓原代细胞。miR-148 a基因敲低后，MM细胞主要分布于G 0/G1期，增殖能力下降。miR-148 a的敲低显著降低了p-NPAT、p-Rb和p-CDC 6的蛋白表达。结论：miR-148 a可通过下调CDKN 1B的表达，抑制CDK活性，促进MM细胞增殖。(C)2016 Elsevier Masson SAS。All rights reserved.
Objective: The aim of this study is to explore the influence of miR-148a on cell proliferation and cell cycle of multiple myeloma (MM) cell line RPMI8226 and the related molecular mechanism.Methods: The expression of miR-148a and CDKN1B in MM cells and primary cells of normal bone marrow were determined by RT-PCR and western blotting. The cell proliferation and cell cycle of miR-148a knockdown MM cells and normal MM cells were determined by flow cytometry. The protein expression of p-NPAT, p-Rb and p-CDC6 was determined in normal and miR-148a knockdown MM cells. Luciferase reported assay was used to explore the relationship between miR-148a and CDKN1B.Results: The level of miR-148a in MM cells was much higher than that in primary cells from healthy bone marrow samples, while the expression of CDKN1B was lower in MM cells. After knockdown of miR-148a, cell cycle mainly distributed at G0/G1 and the proliferation capacity of MM cells decreased. Knockdown of miR-148a significantly reduced protein expression of p-NPAT, p-Rb and p-CDC6. Luciferase reported assay showed that miR-148a could directly target CDKN1B at 30-UTR.Conclusions: High level of miR-148a inhibits CDK activity and promotes the proliferation of MM cells at least partly by downregulating CDKN1B. (C) 2016 Elsevier Masson SAS. All rights reserved.