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中国癌症防治杂志

中国癌症防治杂志 ›› 2024, Vol. 16 ›› Issue (4): 405-411.doi: 10.3969/j.issn.1674-5671.2024.04.04

• 多发性骨髓瘤诊疗专栏 • 上一篇    下一篇

PPP2R5C对多发性骨髓瘤细胞增殖、凋亡和药物敏感性的影响

  

  1. 首都医科大学附属北京同仁医院血液内科
  • 出版日期:2024-08-25 发布日期:2024-08-22
  • 通讯作者: 王亮 E-mail：wangliangtrhos@126.com
  • 基金资助:
    国家自然科学基金面上项目（82170181）；北京市医师科学家培养计划（BJPSTP-2024-01）

Effects of PPP2R5C on proliferation, apoptosis, and drug sensitivity of multiple myeloma cells

  • Online:2024-08-25 Published:2024-08-22

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摘要： 目的 探讨蛋白磷酸酶2调节亚基B'γ（protein phosphatase 2 regulatory subunit B'gamma，PPP2R5C）对多发性骨髓瘤（multiple myeloma，MM）细胞增殖、凋亡和药物敏感性的影响。方法 利用qRT⁃PCR和Western blot检测PPP2R5C在人MM细胞系（MM1S、RPMI⁃8226细胞系）和正常外周血单个核细胞（peripheral blood mononuclear cells，PBMC）中的表达水平。在MM1S细胞中分别转染PPP2R5C干扰siRNA（si⁃PPP2R5C组）及其阴性对照⁃siRNA（si⁃CTRL组）、PPP2R5C过表达质粒（OE⁃PPP2R5C组）及其阴性对照（OE⁃CTRL组），利用CCK⁃8法检测各组细胞的增殖能力，流式细胞术检测细胞凋亡情况。用不同浓度硼替佐米（bortezomib，BTZ）处理si⁃PPP2R5C组和si⁃CTRL组细胞24 h后，采用CCK⁃8法检测细胞活力并计算半数抑制浓度（half maximal inhibitory concentration，IC50）。用1 nmol/L的BTZ干预si⁃PPP2R5C组和si⁃CTRL组细胞24 h后，采用qRT⁃PCR和Western blot检测各组细胞中BCL⁃2和BAX的表达水平，Caspase 3/7 活性检测试剂盒检测Caspase 3/7活性，流式细胞术检测细胞凋亡情况。结果 相较于PBMC细胞，MM1S、RPMI⁃8226细胞在mRNA和蛋白水平上均高表达PPP2R5C（均P<0.001）。与si⁃CTRL组相比，si⁃PPP2R5C组在培养48 h、72 h后细胞增殖活力均受到抑制（均P<0.001），凋亡率明显增加（5.97% vs 14.39%，P<0.001）。与OE⁃CTRL组相比，OE⁃PPP2R5C组在培养48 h、72 h后细胞增殖活力均增强（均P<0.01）。si⁃PPP2R5C组BTZ的IC50值相比si⁃CTRL组显著下降（3.40 nmol/L vs 10.37 nmol/L，P<0.001）。si⁃PPP2R5C+BTZ组相比于Control组和si⁃CTRL+BTZ组，BCL⁃2的mRNA和蛋白水平均下降，而BAX的mRNA及蛋白水平、Caspase3/7活性和细胞凋亡率则增加（均P<0.05）。结论PPP2R5C在MM细胞系中显著高表达，敲低PPP2R5C表达后可抑制MM细胞增殖并促进凋亡，以及增强BTZ的药物敏感性。

关键词: 多发性骨髓瘤, PPP2R5C, 增殖, 凋亡, 耐药

Abstract: Objective To investigate the effects of protein phosphatase 2 regulatory subunit B'gamma (PPP2R5C) on the proliferation, apoptosis and drug sensitivity of multiple myeloma (MM) cells. Methods The qRT⁃PCR and Western blot were used to detect the expression of PPP2R5C in human multiple myeloma cell lines (MM1S and RPMI⁃8226 cell lines) and normal peripheral blood mononuclear cells(PBMC). The MM1S cells were transfected with PPP2R5C interfering siRNA (si⁃PPP2R5C group) and its negative control (si⁃CTRL group), and PPP2R5C overexpressing plasmid (OE⁃PPP2R5C group) and its negative control (OE⁃CTRL group), respectively. Cell proliferation was detected by CCK⁃8 method, and cell apoptosis was detected by flow cytometry. The cells in the si⁃PPP2R5C group and si⁃CTRL group were treated with different concentrations of Bortezomib (BTZ) for 24 h, cell viability was measured by CCK⁃8 method and half maximal inhibitory concentration (IC50) values were calculated. After 1 nmol/L BTZ intervention for 24 h, the expression levels of BCL⁃2 and BAX in the si⁃PPP2R5C and si⁃CTRL groups were detected by qRT⁃PCR and Western blot, and Caspase⁃3/7 activity was detected by Caspase 3/7 activity detection kit, and cell apoptosis was detected by flow cytometry. Results Compared with PBMC cells, PPP2R5C was highly expressed in MM1S and RPMI⁃8226 cells at both mRNA and protein levels (all P<0.001). Compared with the si⁃CTRL group, the cell proliferation activity of si⁃PPP2R5C group was inhibited after 48 h and 72 h of cell culture (all P<0.001), the apoptosis rate was significantly increased (5.97% vs 14.39%, P<0.001). Compared with the OE⁃CTRL group, the cell proliferation activity of OE⁃PPP2R5C group was enhanced at these time points after 48 h and 72 h of cell culture (all P<0.01). The IC50 value of BTZ in the si⁃PPP2R5C group was significantly lower than that in the si⁃CTRL group(3.40 nmol/L vs 10.37 nmol/L, P<0.001). Moreover, compared with the control group and si⁃CTRL+BTZ group, mRNA and protein level of BCL⁃2 in the si⁃PPP2R5C+BTZ group were decreased, while the mRNA and protein level of BAX, Caspase 3/7 activity and cell apoptosis rate were increased (all P<0.05).Conclusions PPP2R5C is highly expressed in MM cell lines. PPP2R5C knockdown can inhibit the cell proliferation and promote apoptosis of MM cells, and increase the drug sensitivity of BTZ.

Key words: Multiple myeloma, PPP2R5C, Proliferation, Apoptosis, Drug resistance

中图分类号: 

  • R733.3