乳腺癌中线粒体功能障碍与CPT1A/ERK信号转导通路共同调节乳腺癌恶性行为的机制研究

doi:10.19401/j.cnki.1007-3639.2024.07.004

摘要/Abstract

摘要：

背景与目的：肉碱棕榈酰转移酶1A（carnitine palmitoyl transferase 1A，CPT1A）的高表达与乳腺癌患者预后较差相关，且能促进线粒体对脂肪酸的利用并最大限度地提高三磷酸腺苷（triphosphate，ATP）产量。然而，CPT1A在乳腺癌转移中的作用仍不清楚。本研究旨在探讨乳腺癌中线粒体功能障碍与CPT1A/细胞外信号调节激酶（extracellular signal-regulated kinase，ERK）信号转导通路共同调节乳腺癌恶性行为的机制。方法：分别使用慢病毒系统和shRNA工具在人乳腺癌细胞系MDA-MB-231和MCF7中过表达或敲低CPT1A，将细胞分为NC组、CPT1A组和shCPT1A组。采用transwell实验检测细胞侵袭能力。采用蛋白质印迹法（Western blot）分析细胞中过氧化物酶体增殖物激活受体γ辅激活因子-1α（peroxisome proliferator-activated receptor γ coactivator-1α，PGC-1α）、ERK1/2和CPT1A蛋白的表达。采用线粒体红染色分析MDA-MB-231和MCF7细胞系的线粒体形态，并通过耗氧率分析线粒体呼吸能力。结果：与表达对照载体的细胞相比，CPT1A过表达导致MCF7细胞和MDA-MB-231细胞侵袭能力增强（P<0.05），shCPT1A敲低导致细胞侵袭能力降低（P<0.05）。与NC组相比，CPT1A组MDA-MB-231和MCF7细胞中线粒体分支长度显著变短（P<0.05），ERK1/2、PGC-1α表达显著增加（P<0.05），shCPT1A组MDA-MB-231和MCF7细胞中线粒体分支长度显著变长（P<0.05），ERK1/2、PGC-1α表达显著降低（P<0.05）。此外，与NC组相比，CPT1A组MDA-MB-231细胞基础和最大呼吸能力以及ATP产量显著增加（P<0.05），而shCPT1A组MDA-MB-231细胞基础和最大呼吸能力以及ATP产量显著降低（P<0.05）。结论：CPT1A激活的ERK1/2-PGC-1α信号转导通路在线粒体分裂介导的乳腺癌细胞转移中发挥重要作用。

关键词: 乳腺癌, 线粒体, 肉碱棕榈酰转移酶1A, 细胞外信号调节激酶, 恶性行为

Abstract:

Background and purpose: The overexpression of carnitine palmitoyl transferase 1A (CPT1A) is related to the poor prognosis of breast cancer, and it can promote the utilization of fatty acids by mitochondria and maximize triphosphate (ATP) production. However, the role of CPT1A in breast cancer metastasis is still unclear. This study aimed to explore the mechanism that mitochondrial dysfunction and CPT1A/extracellular signal-regulated kinase (ERK) signaling pathway in breast cancer jointly regulate the malignant behavior of breast cancer. Methods: The lentivirus system and shRNA tools were used to overexpress or knock down CPT1A in human breast cancer cell lines MDA-MB-231 and MCF7, and the cells were divided into NC group, CPT1A group and shCPT1A group. The invasion ability of cells was detected by transwell assay, and the protein expressions of peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α), ERK1/2 and CPT1A were analyzed by Western blot. The mitochondrial morphology of MDA-MB-231 and MCF7 cell lines was analyzed using mitochondrial red staining, and mitochondrial respiratory capacity was analyzed by oxygen consumption rate. Results: Compared with cells expressing control vector, overexpression of CPT1A resulted in enhanced invasion abilities of MCF7 cells and MDA-MB-231 cells (P<0.05), while knockdown of shCPT1A resulted in decreased invasion abilities of cells (P<0.05). Compared with NC group, the length of mitochondrial branches in MDA-MB-231 and MCF7 cells in CPT1A group was significantly shorter (P<0.05), and the expressions of ERK1/2 and PGC-1α increased significantly (P <0.05). In shCPT1A group, the length of mitochondrial branches in MDA-MB-231 and MCF7 cells increased significantly (P <0.05), and the expressions of ERK1/2 and PGC-1α decreased significantly (P<0.05). In addition, compared with NC group, the cellular basis, maximum respiratory capacity and ATP production of MDA-MB-231 in CPT1A group increased significantly (P <0.05), while the cellular basis, maximum respiratory capacity and ATP production of MDA-MB-231 in shCPT1A group decreased significantly (P<0.05). Conclusion: ERK1/2-PGC-1α activated by CPT1A Signaling pathway plays a key role in mitochondrial division mediated breast cancer cell metastasis.

Key words: Breast cancer, Mitochondria, Carnitine palmitoyl transferase 1A, Extracellular signal-regulated kinase, Malignant behavior

中图分类号:

R737.9

姜丹, 宋国庆, 王晓丹. 乳腺癌中线粒体功能障碍与CPT1A/ERK信号转导通路共同调节乳腺癌恶性行为的机制研究[J]. 中国癌症杂志, 2024, 34(7): 650-658.

JIANG Dan, SONG Guoqing, WANG Xiaodan. Study on the mechanism of mitochondrial dysfunction and CPT1A/ERK signal transduction pathway regulating malignant behavior in breast cancer[J]. China Oncology, 2024, 34(7): 650-658.

图/表 4

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