Rab1A对多发性骨髓瘤细胞系8226增殖和凋亡的影响

doi:10.16098/j.issn.0529-1356.2021.01.009

摘要/Abstract

摘要：

目的探讨Rab1A对多发性骨髓瘤(MM)细胞系8226增殖和凋亡的影响。方法采用siRNA干扰Rab1A表达，将多发性骨髓瘤细胞8226分为空白对照组、阴性对照组和Rab1A siRNA组。其中空白对照组的多发性骨髓瘤细胞8226不做任何处理，阴性对照组的多发性骨髓瘤细胞8226转染阴性对照siRNA。Rab1A siRNA组转染Rab1A特异性siRNA。应用集落形成、CCK-8细胞增殖实验分析Rab1A对多发性骨髓瘤细胞8226增殖的影响。采用流式细胞技术检测多发性骨髓瘤细胞8226的凋亡情况。采用Western blotting和Real-time PCR技术检测Rab1A siRNA对c-Myc、cyclin D1、Bcl-2及Bax表达的影响。结果Rab1A siRNA组多发性骨髓瘤细胞8226的Rab1A mRNA和Rab1A 蛋白的表达显著较阴性对照组下调；集落形成、 CCK-8细胞增殖实验结果显示，Rab1A siRNA可抑制多发性骨髓瘤细胞8226增殖；Rab1A siRNA组多发性骨髓瘤细胞8226的凋亡比例显著较阴性对照组增加（P<0.01）；Rab1A siRNA组c-Myc、cyclin D1和Bcl-2的蛋白表达量显著较阴性对照组下调（P<0.01），Bax蛋白水平的表达量显著较阴性对照组上调（P<0.01）。结论Rab1A可能通过调控c-Myc、cyclin D1、Bcl-2和Bax的表达促进多发性骨髓瘤细胞8226增殖，而RablA siRNA可有效抑制RPMI-8226细胞RablA的表达，从而抑制其增值，并促进凋亡。

关键词: Rab1A, 多发性骨髓瘤细胞, 增殖, 免疫印迹法, 人

Abstract:

Objective To investigate the effect of Rab1A on proliferation and apoptosis of multiple myeloma(MM) cell line 8226. Methods The siRNA interference was used to knockdown Rab1A gene. The multiple myeloma cell line 8226 was divided into blank control group, negative control group and Rab1A siRNA group. In the blank control group, the multiple myeloma cells were not treated. Multiple myeloma cells 8226 in the negative control group were transfected with negative control siRNA. The Rab1A siRNA group was transfected with Rabl A-targeted siRNA. The effect of Rab1A on multiple myeloma cell 8226 proliferation was analyzed by colony forming test and cell counting kit-8 (CCK-8) assay. The apoptosis of multiple myeloma cell 8226 was detected by flow cytometry. Western blotting and Real-time PCR were used to observe the effect of Rab1A siRNA on the expression of c-Myc, cyclin D1, Bcl-2 and Bax. Results The expressions of Rab1A mRNA and Rab1A protein in the Rab1A siRNA group were significantly down-regulated compared with those in the negative control group. The result of colony formation and CCK-8 assay showed that Rab1A siRNA inhibit the proliferation of multiple myeloma cells 8226. The early and late apoptosis ratio of multiple myeloma cell 8226 in Rab1A siRNA group increased significantly compared with the negative control group (P<0.05). The expression of cyclin D1 and Bcl-2 in the Rab1A siRNA group were significantly down-regulated compared with the negative control group (P<0.05), and the expression of c-Myc and Bax were significantly up-regulated compared with the negative control group (P<0.05). ConclusionRab1A may promote the proliferation of multiple myeloma cells 8226 by regulating the expression of c-Myc, cyclin D1, Bcl-2 and Bax, while RablA siRNA can effectively inhibit the expression of RablA in rpmi-8226 cells, thereby inhibiting its proliferation and promoting apoptosis.

Key words: Rab1A, Multiple myeloma cell, Proliferation, Western blotting, Human

中图分类号:

R733.3

吴含王秀红吴婷婷杨粟. Rab1A对多发性骨髓瘤细胞系8226增殖和凋亡的影响[J]. 解剖学报, 2021, 52(1): 60-66.

WU Han WANG Xiu-hong WU Ting-ting YANG Su. Effects of Rab1A on proliferation and apoptosis of multiple myeloma cell line 8226[J]. Acta Anatomica Sinica, 2021, 52(1): 60-66.

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