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中国癌症防治杂志

中国癌症防治杂志 ›› 2022, Vol. 14 ›› Issue (4): 378-387.doi: 10.3969/j.issn.1674-5671.2022.04.04

• 基础研究 • 上一篇    下一篇

PGRMC1在激素治疗中促进乳腺癌细胞增殖的机制研究 

  

  1. 首都医科大学附属北京妇产医院/北京妇幼保健院内分泌科 
  • 出版日期:2022-08-25 发布日期:2022-09-02
  • 通讯作者: 阮祥燕 E-mail：ruanxiangyan@ccmu.edu.cn
  • 基金资助:
    北京市教育委员会科技发展计划一般项目（KM202010025009）；北京市医院管理中心“青苗”计划专项经费资助（QML20181401）；国家自然科学基金项目（81671411）；北京市自然科学基金项目（7162062）

Mechanism of PGRMC1 promoting breast cancer cell proliferation during hormone therapy

  • Online:2022-08-25 Published:2022-09-02

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摘要： 目的 探讨孕激素受体膜组分1（progesterone receptor membrane components 1，PGRMC1）在激素治疗中促进乳腺癌细胞增殖的作用机制。 方法 雌激素受体（estrogen receptor，ER）阳性的乳腺癌MCF7细胞经雌孕激素处理后，用过表达PGRMC1、敲降ER抑制因子PHB1（shPHB1-1和shPHB1-2）及其相应阴性对照的慢病毒液进行感染。采用免疫共沉淀联合质谱分析及GST pull-down实验检测PGRMC1与PHB复合体（PHB1和PHB2）的相互作用，免疫印迹法和RT-qPCR检测PHB1、PHB2和PGRMC1的表达情况，CCK-8和EDU实验检测细胞的增殖能力，流式细胞术检测细胞周期情况，RT-qPCR检测ER信号通路下游靶基因THBS1、CXCL12和GREB1的表达情况。结果 免疫共沉淀联合质谱分析发现，PHB1和PHB2均存在于PGRMC1的免疫共沉淀组分中；GST pull-down鉴定体外条件下PGRMC1与PHB复合体存在直接相互作用。与相应对照组相比，过表达PGRMC1和下调PHB1的乳腺癌细胞增殖速度均明显加快（均P<0.05），S期和G2/M期阳性细胞比例均明显增加（均P<0.05），且能够显著促进ER信号通路下游靶基因THBS1、CXCL12和GREB1的表达（均P<0.01）。PGRMC1过表达后MCF7细胞中PHB1与ER的相互作用减弱。下调PHB1后再过表达PGRMC1的细胞周期中，S期和G2/M期阳性细胞比例与单独下调PHB1相比无明显变化（均P>0.05）。结论 PGRMC1可能通过与PHB复合体结合，并解除后者对ER信号通路的抑制作用，从而促进ER信号通路的活化和下游靶基因的表达，加速激素刺激条件下乳腺癌细胞的恶性增殖。

关键词: 乳腺癌, 孕激素受体膜组分1, 雌激素受体, ER抑制因子1, ER抑制因子2

Abstract: Objective To investigate the mechanism of progesterone receptor membrane components 1(PGRMC1) promoting the proliferation of breast cancer cells during hormone therapy. Methods Estrogen receptor (ER) -positive breast cancer MCF7 cells were treated with estrogen and progesterone, and then infected with chronic disease venom overexpressing PGRMC1, knockdown ER prohibitin(shPHB1-1 and shPHB1-2) and their corresponding negative controls. The interaction between PGRMC1 and PHB complex (PHB1 and PHB2) was detected by co-immunoprecipitation combined with mass spectrometry and GST pull-down assay. The expressions of PHB1, PHB2 and PGRMC1 were detected by Western blot and RT-qPCR. The cell proliferation ability was detected by CCK-8 and EDU assay. Flow cytometry was used to detect the cell cycle, and RT-qPCR was used to detect the expression of downstream target genes of ER signaling pathway, including THBS1, CXCL12 and GREB1. Results Co-immunoprecipitation combined with mass spectrometry analysis showed that both PHB1 and PHB2 were present in the co-immunoprecipitation fraction of PGRMC1. GST pull-down identified a direct interaction between PGRMC1 and PHB complex in vitro. Compared with the control group, the proliferation rate of breast cancer cells overexpressing PGRMC1 and down-regulating PHB1 was significantly accelerated (all P<0.05), the proportions of positive cells in S and G2/M phases were significantly increased (all P<0.05), and the expressions of THBS1, CXCL12 and GREB1 in the downstream target genes of ER signaling pathway were significantly promoted (all P<0.01). The interaction between PHB1 and ER in MCF7 cells was attenuated after PGRMC1 overexpression. The proportion of positive cells in S and G2/M phases after down-regulating PHB1 and then overexpressing PGRMC1 had no significant change compared with that of down-regulating PHB1 alone (all P>0.05). Conclusions PGRMC1 may relieve the inhibitory effect of PHB complex on the ER signaling pathway by binding to PHB complex, thereby promoting the activation of the ER signaling pathway and the expressions of downstream target genes, and accelerating the malignant proliferation of breast cancer cells under hormone stimulation.

Key words: Breast cancer, Progesterone receptor membrane components 1, Estrogen receptor, Prohibitin 1, Prohibitin 2

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