CENPA 敲低通过降低 PLA2R1 启动子甲基化和调节 PLA2R1/HHEX 轴来抑制乳腺癌细胞的进展和肿瘤生长。
CENPA knockdown restrains cell progression and tumor growth in breast cancer by reducing PLA2R1 promoter methylation and modulating PLA2R1/HHEX axis.
机构信息
Department of Surgery, The First Affiliated Hospital of Jinzhou Medical University, Jinzhou, 121000, Liaoning, China.
Department of Rheumatology, The First Affiliated Hospital of Jinzhou Medical University, Jinzhou, 121000, Liaoning, China.
出版信息
Cell Mol Life Sci. 2024 Jan 12;81(1):27. doi: 10.1007/s00018-023-05063-5.
BACKGROUND
Breast cancer is a lethal malignancy affecting females worldwide. It has been reported that upregulated centromere protein A (CENPA) expression might indicate unfortunate prognosis and can function as a prognostic biomarker in breast cancer. This study aimed to investigate the accurate roles and downstream mechanisms of CENPA in breast cancer progression.
METHODS
CENPA protein levels in breast cancer tissues and cell lines were analyzed by Western blot and immunohistochemistry assays. We used gain/loss-of-function experiments to determine the potential effects of CENPA and phospholipase A2 receptor (PLA2R1) on breast cancer cell proliferation, migration, and apoptosis. Co-IP assay was employed to validate the possible interaction between CENPA and DNA methyltransferase 1 (DNMT1), as well as PLA2R1 and hematopoietically expressed homeobox (HHEX). PLA2R1 promoter methylation was determined using methylation-specific PCR assay. The biological capabilities of CENPA/PLA2R1/HHEX axis in breast cancer cells was determined by rescue experiments. In addition, CENPA-silenced MCF-7 cells were injected into mice, followed by measurement of tumor growth.
RESULTS
CENPA level was prominently elevated in breast cancer tissues and cell lines. Interestingly, CENPA knockdown and PLA2R1 overexpression both restrained breast cancer cell proliferation and migration, and enhanced apoptosis. On the contrary, CENPA overexpression displayed the opposite results. Moreover, CENPA reduced PLA2R1 expression through promoting DNMT1-mediated PLA2R1 promoter methylation. PLA2R1 overexpression could effectively abrogate CENPA overexpression-mediated augment of breast cancer cell progression. Furthermore, PLA2R1 interacted with HHEX and promoted HHEX expression. PLA2R1 knockdown increased the rate of breast cancer cell proliferation and migration but restrained apoptosis, which was abrogated by HHEX overexpression. In addition, CENPA silencing suppressed tumor growth in vivo.
CONCLUSION
CENPA knockdown restrained breast cancer cell proliferation and migration and attenuated tumor growth in vivo through reducing PLA2R1 promoter methylation and increasing PLA2R1 and HHEX expression. We may provide a promising prognostic biomarker and novel therapeutic target for breast cancer.
背景
乳腺癌是一种影响全球女性的致命恶性肿瘤。据报道，上调着丝粒蛋白 A（CENPA）的表达可能预示着不良预后，并可作为乳腺癌的预后生物标志物。本研究旨在探讨 CENPA 在乳腺癌进展中的准确作用和下游机制。
方法
通过 Western blot 和免疫组织化学检测分析乳腺癌组织和细胞系中的 CENPA 蛋白水平。我们使用增益/失能实验来确定 CENPA 和磷脂酶 A2 受体（PLA2R1）对乳腺癌细胞增殖、迁移和凋亡的潜在影响。采用 co-IP 实验验证 CENPA 与 DNA 甲基转移酶 1（DNMT1）以及 PLA2R1 与血源性同源盒（HHEX）之间的可能相互作用。采用甲基化特异性 PCR 检测 PLA2R1 启动子甲基化。通过拯救实验确定 CENPA/PLA2R1/HHEX 轴在乳腺癌细胞中的生物学功能。此外，用 CENPA 沉默的 MCF-7 细胞注射小鼠，然后测量肿瘤生长。
结果
CENPA 水平在乳腺癌组织和细胞系中明显升高。有趣的是，CENPA 敲低和 PLA2R1 过表达均抑制乳腺癌细胞增殖和迁移，并增强凋亡。相反，CENPA 过表达则呈现相反的结果。此外，CENPA 通过促进 DNMT1 介导的 PLA2R1 启动子甲基化降低 PLA2R1 的表达。PLA2R1 过表达可以有效消除 CENPA 过表达介导的乳腺癌细胞进展的增强。此外，PLA2R1 与 HHEX 相互作用并促进 HHEX 表达。PLA2R1 敲低增加了乳腺癌细胞增殖和迁移的速度，但抑制了凋亡，而过表达 HHEX 则可以消除这种作用。此外，CENPA 沉默抑制体内肿瘤生长。
结论
CENPA 敲低通过减少 PLA2R1 启动子甲基化和增加 PLA2R1 和 HHEX 表达，抑制乳腺癌细胞增殖和迁移，并减弱体内肿瘤生长。我们可能为乳腺癌提供了一个有前途的预后生物标志物和新的治疗靶点。
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