LSD1抑制剂对前列腺癌细胞体外生物学行为的调控

Effect of LSD1 inhibitors on regulating the biological behaviors of prostate cancer cells in vitro

目的研究赖氨酸特异性去甲基化酶-1(lysine specific demethylase 1,LSD1)抑制剂优降宁对人雄激素非依赖性前列腺癌细胞增殖、迁移以及上皮间质转化等体外生物学行为的影响。方法分别用不同浓度的优降宁(0、1、3 mmol/L)处理肿瘤细胞0~72 h,使用CCK8法检测细胞存活率。选用0、3 mmol/L优降宁处理48 h,用流式细胞仪检测比较细胞的凋亡以及细胞周期分布情况,用划痕实验及Transwell小室迁移实验比较细胞的运动能力,用Western-blot实验比较上皮间质转化过程中的关键基因的蛋白表达差异,最终用RT-q PCR进一步验证m RNA水平的变化,以阐明LSD1抑制剂优降宁对前列腺癌细胞的影响。结果 1、3 mmol/L的优降宁均能够显著降低DU145细胞的存活率以及集落形成数,以3 mmol/L剂量处理细胞48 h最为显著。3 mmol/L优降宁处理48 h后能显著增加前列腺癌细胞的凋亡率,使得停留在G2期的细胞明显增加。处理组划痕愈合速率明显减慢,相同时间穿过小室的细胞数明显减少。蛋白水平和m RNA水平结果一致表明上皮间质转化过程受到显著抑制。结论 LSD1抑制剂能够通过促进细胞凋亡、阻滞细胞周期显著抑制前列腺癌细胞的增殖,减弱肿瘤细胞的迁移运动能力,并且显著抑制肿瘤细胞上皮间质转化过程,LSD1可成为前列腺癌治疗的潜在靶点。

Objective To investigate the effect oflysine-specific demethylase-1 （LSD1） inhibitor Eutonyl on the proliferation, migration and epithelial-mesenchymal transition and other in vitro biological behaviors of androgen-independent human prostate cancer cells. Methods Tumor cells were treated with different concentrations of Eutonyl （0, 1, 3 mmol/L） for 0-72 h. The cell viability was measured by CCK8 assay. After treatment with 0 and 3 mmol/L Eutonyl for 48 h, the cell apoptosis and the distribution of cell cycle were compared by flow cytometry, and the cell viability was detected by the scratch test and Transwell chamber test. Western-blot was adopted to measure the protein expression of key genes in the process of epithelial mesenchymal transition. RT-qPCR was performed to further verify the changes in mRNA levels to elucidate the effect of LSD1 inhibitor Eutonyl on prostate cancer cells. Results Eutonyl at a dose of 1 and 3 mmol/L could significantly reduce the survival rate and colony formation of DU145 cells. The most significant effect was obtained after 3 mmol/L Eutonyl treatment for 48 h. Eutonyl treatment at a dose of 3 mmol/L for 48 h could significantly increase the apoptosis rate of prostate cancer cells and evidently increased the quantity of cells in the G2 phase. The healing rate of scratch test was significantly slowed and the quantity of cells passing through the chamber was significantly reduced in thetreatment groups. The results at the protein and mRNA levels were consistent, suggesting that the process of epithelial-mesenchymal transition was significantly inhibited. Conclusion LSD1 inhibitor can significantly inhibit the proliferation of prostate cancer cells by promoting cell apoptosis and arresting cell cycle, weakening the migration ability of tumor cells, and significantly suppress the epithelial-mesenchymal transition process. LSD1 may be a potential therapeutic target for clinical treatment of prostate cancer.