Zhang Xu-Yang, Hao Pan, Wang Jun-Wei, Zhao Wen, Liu Hong-Min, He Peng-Xing
Institute of Drug Discovery & Development, School of Pharmaceutical Sciences, State Key Laboratory of Esophageal Cancer Prevention and Treatment, Key Laboratory of Advanced Drug Preparation Technologies, Ministry of Education of China, Zhengzhou University, 450001, Zhengzhou, China.
Mol Biol Rep. 2023 Jan;50(1):507-516. doi: 10.1007/s11033-022-07960-7. Epub 2022 Nov 9.
Lysine-Specific Demethylase 1 (LSD1) inhibitors have been developed and reached the clinic, but its effect in combination with cytotoxic chemotherapy is unclear. Here, we investigated the anti-tumor effect of LSD1 inhibitor GSK-LSD1 and its anti-tumor effect with the DNA damage drug doxorubicin (DOX) in gastric cancer (GC) cells.
Cells were treated with different concentrations of GSK-LSD1 to examine the anti-tumor effect versus cell viability by MTT and cell cycle arrest by flow cytometry. To explore whether LSD1 inhibitors can increase the anti-tumor effect of DNA damage drugs, cells were treated with DOX for 48 h after pretreatment with GSK-LSD1 for 48 h. Cell viability was detected by MTT and apoptosis-related proteins were examined by Western blot. Furthermore, anti-tumor efficacy of combination GSK-LSD1 with DOX was also measured in MGC-803 xenografts model in nude mice.
The results showed that LSD1 was highly expressed in GC cell lines. Inhibition of LSD1 has a weak effect on cell viability and cell cycle. Moreover, LSD1 inhibitors pretreatment could significantly increase the anti-tumor effect of DOX. Further study found that inhibition of LSD1 can significantly enhance DOX-induced the apoptosis, accompanied by down-regulation of antiapoptotic Bcl-2 expression and up-regulation of proapoptotic Bax expression. We also confirmed that inhibition of LSD1 can sensitize the anti-tumor effect of DOX in vivo.
Our findings suggest that the LSD1 inhibitor GSK-LSD1 has a weak inhibitory effect on the viability and cell cycle of GC cells, but can enhance the sensitivity of DOX.
赖氨酸特异性去甲基化酶1（LSD1）抑制剂已研发并进入临床，但它与细胞毒性化疗联合使用的效果尚不清楚。在此，我们研究了LSD1抑制剂GSK-LSD1在胃癌（GC）细胞中的抗肿瘤作用及其与DNA损伤药物阿霉素（DOX）联合使用的抗肿瘤作用。
用不同浓度的GSK-LSD1处理细胞，通过MTT法检测其对细胞活力的抗肿瘤作用，并通过流式细胞术检测细胞周期阻滞情况。为探讨LSD1抑制剂是否能增强DNA损伤药物的抗肿瘤作用，先用GSK-LSD1预处理细胞48小时，再用DOX处理48小时。通过MTT法检测细胞活力，并用蛋白质免疫印迹法检测凋亡相关蛋白。此外，还在裸鼠的MGC-803异种移植模型中检测了GSK-LSD1与DOX联合使用的抗肿瘤疗效。
结果显示，LSD1在GC细胞系中高表达。抑制LSD1对细胞活力和细胞周期的影响较弱。此外，LSD1抑制剂预处理可显著增强DOX的抗肿瘤作用。进一步研究发现，抑制LSD1可显著增强DOX诱导的细胞凋亡，同时伴随着抗凋亡蛋白Bcl-2表达的下调和促凋亡蛋白Bax表达的上调。我们还证实，抑制LSD1可使DOX在体内的抗肿瘤作用敏感化。
我们的研究结果表明，LSD1抑制剂GSK-LSD1对GC细胞的活力和细胞周期具有较弱的抑制作用，但可增强DOX的敏感性。
