Su L Z, Chen J, Li X, Ji P
Department of Oral and Maxillofacial Surgery, Stomatological Hospital of Chongqing Medical University, Chongqing 401120, China.
Chongqing Key Laboratory of Oral Diseases and Biomedical Sciences, Chongqing 401120, China.
Beijing Da Xue Xue Bao Yi Xue Ban. 2020 Oct 18;52(5):902-906. doi: 10.19723/j.issn.1671-167X.2020.05.018.
To investigate the effects of salinomycin on the proliferation and apoptosis of oral squamous carcinoma cells and to further understand the mechanisms of these effects.
The human oral squamous carcinoma cell line CAL-27 was cultured in different concentrations of salinomycin and cisplatin. After co-culture with 0, 1, 2, 4, 8, 16 and 32 μmol/L salinomycin or 0, 1.25, 2.5, 5, 10, 20, 40 and 80 μmol/L cisplatin for 24 hours and 48 hours, the proliferation of oral squamous carcinoma cells were detected by cell counting kit-8(CCK-8) assay. After being exposed to 0, 2, 4, 8 μmol/L salinomycin and 0, 5, 10, 20 μmol/L cisplatin for 48 hours, the cell cycle of oral squamous carcinoma cells was detected by flow cytometry assay, and Western blot analysis was performed to analyze the expressions of cysteine-containing aspartate-specific proteases-3(Caspase-3), cysteine-containing aspartate-specific proteases-9(Caspase-9), poly ADP-ribose polymerase (PARP), protein kinase B (Akt) and phosphorylated protein kinase B (p-Akt) protein in oral squamous carcinoma cells.
Both salinomycin and cisplatin significantly inhibited the proliferation of oral squamous cell carcinoma CAL-27 cells in a time- and dose-dependent manner. However, compared with the first-line chemotherapeutic drug cisplatin, salinomycin showed stronger anti-proliferation activity in oral squamous carcinoma cells than cisp-latin ( < 0.001). After being exposed to 8 μmol/L salinomycin, CAL-27 cells exhibited markedly higher proportion in quiescent/ first gap phases (40.40%±1.99% . 64.46%±0.90%, < 0.05), and had a significantly lower proportion in synthesis phases and second gap / mitosis phases (24.32%±2.30% . 18.73%±0.61%, < 0.05; 35.01%±1.24% . 16.54%±1.31%, < 0.05) compared with the dimethyl sulfoxide control group; moreover cisplatin didn't show cell-cycle specific effect on CAL-27. Western blot proved that salinomycin could up-regulate the expressions of Caspase-3 and Caspase-9 protein in oral squamous cell carcinoma CAL-27 cells ( < 0.05). At the same time, the levels of PARP, Akt and p-Akt protein were down-regulated ( < 0.05).
Compared with cisplatin, salinomycin has a better inhibitory effect on the proliferation of oral squamous carcinoma cells and blocks the cell cycle process at the quiescent / first gap phase. At the same time, salinomycin could trigger apoptosis of oral squamous carcinoma cells and the mechanism is associated with the Akt/p-Akt signaling pathway.
研究盐霉素对口腔鳞状癌细胞增殖和凋亡的影响，并进一步了解其作用机制。
将人口腔鳞状癌细胞系CAL-27培养于不同浓度的盐霉素和顺铂中。分别与0、1、2、4、8、16和32 μmol/L盐霉素或0、1.25、2.5、5、10、20、40和80 μmol/L顺铂共培养24小时和48小时后，采用细胞计数试剂盒-8（CCK-8）法检测口腔鳞状癌细胞的增殖情况。将细胞分别暴露于0、2、4、8 μmol/L盐霉素和0、5、10、20 μmol/L顺铂中48小时后，采用流式细胞术检测口腔鳞状癌细胞的细胞周期，并通过蛋白质免疫印迹分析检测口腔鳞状癌细胞中含半胱氨酸的天冬氨酸蛋白水解酶-3（Caspase-3）、含半胱氨酸的天冬氨酸蛋白水解酶-9（Caspase-9）、聚ADP核糖聚合酶（PARP）、蛋白激酶B（Akt）和磷酸化蛋白激酶B（p-Akt）蛋白的表达。
盐霉素和顺铂均能以时间和剂量依赖性方式显著抑制口腔鳞状细胞癌CAL-27细胞的增殖。然而，与一线化疗药物顺铂相比，盐霉素在口腔鳞状癌细胞中显示出比顺铂更强的抗增殖活性（P<0.001）。暴露于8 μmol/L盐霉素后，CAL-27细胞在静止/第一间隙期的比例显著升高（40.40%±1.99%、64.46%±0.90%，P<0.05），而在合成期和第二间隙/有丝分裂期的比例显著降低（24.32%±2.30%、18.73%±0.61%，P<0.05；35.01%±1.24%、16.54%±1.31%，P<0.05），与二甲基亚砜对照组相比；此外，顺铂对CAL-27细胞未显示出细胞周期特异性作用。蛋白质免疫印迹证明，盐霉素可上调口腔鳞状细胞癌CAL-27细胞中Caspase-3和Caspase-9蛋白的表达（P<0.05）。同时，PARP、Akt和p-Akt蛋白水平下调（P<0.05）。
与顺铂相比，盐霉素对口腔鳞状癌细胞增殖的抑制作用更好，并将细胞周期进程阻滞在静止/第一间隙期。同时，盐霉素可引发口腔鳞状癌细胞凋亡，其机制与Akt/p-Akt信号通路有关。
