百乐明TCP对卵巢癌细胞增殖、凋亡和自噬的影响

Lysine-Specific Demethylase 1(LSD1) Inhibitor TCP Induces Apoptosis and Autophagy via the AKT/mTOR Pathway in SKOV3 Ovarian Cancer Cells

目的强内心百乐明(tranylcypromine,TCP)是在卵巢癌中最有效的LSD1抑制剂之一,被证明可以抑制卵巢癌细胞的增殖,但是其中更深入的分子生物学机制目前尚无研究。本研究的目的在于讨论TCP对卵巢癌生物学功能的影响。方法本研究采用了CCK-8法检测了细胞的增殖能力,流式细胞术检测了细胞的凋亡能力,western blot法检测了细胞中自噬的发生和mTOR通路的活性。结果我们的研究证明了在卵巢癌细胞中,应用浓度为100μM LSD1抑制剂TCP处理细胞48h可以诱发早期凋亡和晚期凋亡的发生,Bcl-2的表达上升,Bax的表达下降,这提示我们TCP可以诱发卵巢癌细胞凋亡的发生。当应用TCP处理SKOV3细胞12h后开始发生自噬,而在24h和48h后自噬效果最为明显。而TCP处理卵巢癌细胞24h和48h后p-mTOR、p-P70S6K蛋白的表达水平明显下降。结论 LSD1抑制剂TCP可以抑制SKOV3细胞的增殖,诱发细胞凋亡和自噬能力,同时抑制mTOR/p70S6K细胞信号转导通路的活性。

Objective TCP is one of the most potent LSD1 inhibitors,which can inhibit ovarian cancer cells viability. This study aimed to investigate the mechanism of the anti-cancer properties of TCP in SKOV3 ovarian cells. Methods Cell viability was measured by Cell Counting Kit-8（CCK-8） assay. Cellular apoptosis was evaluated by flowcytometric analysis using Annexin-V/PI staining methods. Western blotting was performed for analyzing the Bax,Bcl-2,mTOR,p-mTOR,p62,LC3-I,LC3-II protein expressions. Results Our results showed that the proportion of early apoptotic and late apoptotic cells increased significantly for cells treated with TCP at a concentration of 100 μMfor 48 h. Treatment of TCP in SKOV3 cells resulted in upregulation of Bax and downregulation of Bcl-2 in a time-dependent manner,indicating that TCP can induce apoptosis in SKOV3 cells. There was a downward trend in the expression of p62 when the SKOV3 cells were treated with 100 μm TCP for 12 h,24 h and 48 h. The conversion of LC3-I to LC3-II was increased significantly at 24 h and 48 h. TCP treatment decreased the levels of phosphorylated mTOR. Conclusion TCP inhibits SKOV3 cells viability and induces apoptosis and autophagy. The mTOR signaling pathway was found to be affected by TCP.