Zong Shoukai, Dai Wei, Fang Wencheng, Guo Xiangting, Wang Kai
People's Hospital of Rizhao, Rizhao, China.
Biosci Rep. 2020 May 27. doi: 10.1042/BSR20201302.
This study aimed to investigate the effect of SIK2 on cisplatin resistance induced by aerobic glycolysis in breast cancer cells and its potential mechanism. qRt-PCR and Western blot were used to detect SIK2 mRNA and protein levels, and cisplatin (DDP) resistant cell lines of breast cancer cells were established. Viability was measured and evaluated via CCK-8, cell invasion capability was evaluated via Transwell, and apoptosis rate was assessed via Flow cytometry. The glycolysis level was evaluated by measuring glucose consumption and lactic acid production. The protein levels of p-PI3K, p- protein kinase B (Akt) and p-mTOR were determined by western blot. SIK2 was highly expressed in breast cancer tissues and cells compared with adjacent tissues and normal human breast epithelial cells, and it had higher diagnostic value for breast cancer. Silencing SIK2 expression can inhibit proliferation and invasion of breast cancer cells and induce their apoptosis. In addition, SIK2 knockdown inhibits glycolysis, reverses the resistance of drug-resistant cells to cisplatin, and inhibits PI3K/AKT/mTOR signaling pathway. When LY294002 was used to inhibit PI3K/AKT/mTOR signaling pathway, the effect of pcDNA3.1-SIK2 on aerobic glycolysis of breast cancer cells could be reversed. SIK2 can promote cisplatin resistance caused by aerobic glycolysis of breast cancer cells through PI3K/AKT/mTOR signaling pathway, which may be a new target to improve cisplatin resistance of breast cancer cells.
本研究旨在探讨盐诱导激酶2（SIK2）对乳腺癌细胞有氧糖酵解诱导的顺铂耐药性的影响及其潜在机制。采用qRt-PCR和蛋白质免疫印迹法检测SIK2 mRNA和蛋白水平，并建立乳腺癌细胞的顺铂（DDP）耐药细胞系。通过CCK-8法检测并评估细胞活力，通过Transwell法评估细胞侵袭能力，通过流式细胞术评估细胞凋亡率。通过测量葡萄糖消耗和乳酸生成来评估糖酵解水平。通过蛋白质免疫印迹法测定磷酸化磷脂酰肌醇-3激酶（p-PI3K）、磷酸化蛋白激酶B（Akt）和磷酸化哺乳动物雷帕霉素靶蛋白（p-mTOR）的蛋白水平。与癌旁组织和正常人乳腺上皮细胞相比，SIK2在乳腺癌组织和细胞中高表达，对乳腺癌具有较高的诊断价值。沉默SIK2表达可抑制乳腺癌细胞的增殖和侵袭并诱导其凋亡。此外，敲低SIK2可抑制糖酵解，逆转耐药细胞对顺铂的耐药性，并抑制PI3K/AKT/mTOR信号通路。当使用LY294002抑制PI3K/AKT/mTOR信号通路时，pcDNA3.1-SIK2对乳腺癌细胞有氧糖酵解的影响可被逆转。SIK2可通过PI3K/AKT/mTOR信号通路促进乳腺癌细胞有氧糖酵解引起的顺铂耐药性，这可能是提高乳腺癌细胞顺铂耐药性的新靶点。
