Liao Linhong, Cheng Hui, Liu Shusong
Department of Pathology, Ganzhou Maternal and Child Health Hospital, Ganzhou, Jiangxi 341000, P.R. China.
Department of Emergency, Ganzhou People's Hospital, Ganzhou, Jiangxi 341000, P.R. China.
Oncol Lett. 2022 Jul 19;24(3):317. doi: 10.3892/ol.2022.13438. eCollection 2022 Sep.
Breast cancer is one of the most frequently diagnosed types of cancer worldwide. The present study aimed to investigate the role and underlying regulatory mechanism of non-structural maintenance of chromosome condensin I complex subunit H (NCAPH) in the malignant progression and cisplatin (DDP) resistance of breast cancer cells. Therefore, the mRNA and protein expression levels of NCAPH were first determined in breast cancer cells via reverse transcription-quantitative PCR and western blotting. Furthermore, following transfection of NCAPH interference plasmids, the effect of NCAPH knockdown on cell proliferation, migration, invasion were also assessed using CCK-8, wound healing and Transwell assays. Apoptosis was evaluated using TUNEL assay, and western blotting was performed in breast cancer cells and DDP-resistant breast cancer cells. The association between NCAPH and its downstream target, aurora kinase B (AURKB), was verified using bioinformatic analysis and the co-immunoprecipitation assay. Furthermore, the effect of AURKB overexpression on the aforementioned processes and the Akt/mTOR signaling pathway were also assessed. The results demonstrated that NCAPH mRNA and protein expression levels were significantly upregulated in breast cancer cells, whereas NCAPH knockdown significantly attenuated the proliferation, migration and invasion of breast cancer cells. NCAPH silencing also exacerbated the apoptosis of DDP-resistant breast cancer cells. AURKB mRNA and protein expression levels were also significantly upregulated in MCF-7 cells, whereas its overexpression significantly reversed the effects of NCAPH knockdown on breast cancer cells and the Akt/mTOR signaling pathway. Overall, NCAPH knockdown significantly downregulated AURKB mRNA and protein expression levels to block the Akt/mTOR signaling pathway and inhibited breast cancer cell proliferation, migration, invasion, and aggravate DDP-resistant breast cancer cell apoptosis, indicating that NCAPH may serve as a promising therapeutic target for breast cancer.
乳腺癌是全球最常被诊断出的癌症类型之一。本研究旨在探讨染色体凝聚素I复合体亚基H（NCAPH）在乳腺癌细胞恶性进展及顺铂（DDP）耐药中的作用及潜在调控机制。因此，首先通过逆转录定量PCR和蛋白质印迹法测定乳腺癌细胞中NCAPH的mRNA和蛋白质表达水平。此外，转染NCAPH干扰质粒后，还使用CCK-8、伤口愈合和Transwell实验评估NCAPH敲低对细胞增殖、迁移、侵袭的影响。使用TUNEL实验评估细胞凋亡，并在乳腺癌细胞和耐DDP乳腺癌细胞中进行蛋白质印迹分析。通过生物信息学分析和免疫共沉淀实验验证NCAPH与其下游靶点极光激酶B（AURKB）之间的关联。此外，还评估了AURKB过表达对上述过程及Akt/mTOR信号通路的影响。结果表明，乳腺癌细胞中NCAPH的mRNA和蛋白质表达水平显著上调，而敲低NCAPH可显著减弱乳腺癌细胞的增殖、迁移和侵袭。沉默NCAPH还加剧了耐DDP乳腺癌细胞的凋亡。MCF-7细胞中AURKB的mRNA和蛋白质表达水平也显著上调，而过表达AURKB可显著逆转敲低NCAPH对乳腺癌细胞及Akt/mTOR信号通路的影响。总体而言，敲低NCAPH可显著下调AURKB的mRNA和蛋白质表达水平，阻断Akt/mTOR信号通路，抑制乳腺癌细胞增殖、迁移、侵袭，并加剧耐DDP乳腺癌细胞凋亡，表明NCAPH可能是乳腺癌一个有前景的治疗靶点。
