摘要
目的:研究新型精胺氧化酶(SMO)小分子抑制剂SI-4650对人卵巢癌SKVO-3细胞增殖和上皮细胞间质化影响及其分子机制。方法:体外培养SKVO-3细胞,以未加药作为对照组,30、60μmol/L SI-4650处理48 h细胞为实验组,每组设3个复孔,检测SI-4650对SKVO-3细胞内SMO的酶活性及多胺含量、酶促反应产物活性氧水平的影响,对细胞增殖、周期、线粒体膜电位的作用,以及对细胞凋亡、侵袭和迁移能力的影响,同时检测凋亡相关蛋白Bax、Bcl-2、Caspase3以及上皮细胞间质化(EMT)相关蛋白E-Cad、N-Cad、Vimentin、MMP-2、MMP-9表达。结果:与对照组相比,实验组SKVO-3细胞中SI-4650浓度增加,明显抑制SKVO-3细胞内SMO酶活性(P<0.01)、减少SMO酶促反应产物活性氧水平(P<0.01)、降低细胞内总多胺含量(P<0.01)。SI-4650高效抑制SKVO-3细胞生长(P<0.01),实验组细胞生长抑制率分别为32.27%、47.31%;将SKVO-3细胞阻滞在S期(P<0.01),实验组细胞Bax表达和Caspase3活化剪切增加,Bcl-2表达减少,线粒体膜电位下降,凋亡细胞比率增加至31.41%、43.51%;同时,实验组E-cad表达显著增加,N-cad、Vimentin表达均明显减少,合成分泌MMP-2、MMP-9减少,干扰了SKVO3细胞EMT进程,降低肿瘤细胞侵袭、迁移能力(P<0.01)。结论:SI-4650对人卵巢癌SKVO-3细胞具有杀伤和抑制肿瘤细胞侵袭、迁移的抗肿瘤活性,其机制可能与干扰多胺代谢、诱导细胞S周期阻滞和线粒体途径细胞凋亡以及干扰细胞EMT进程相关。
Objective:To investigate the effects of SI-4650,a novel small molecule inhibitor of spermine oxidase(SMO),on the proliferation and epithelial mesenchymal transformation(EMT)of human ovarian cancer SKVO-3 cells as well as its underlying molecular mechanisms.Methods:SKVO-3 cells treated with 0μmol/L SI-4650 were used as control group,SKVO-3 cells treated with 30,60μmol/L SI-4650 were used as experimental group.The effects of SI-4650 on the activity of SMO,the polyamine contents and the cellular reactive oxygen species(ROS)were detected.Cell proliferation,cell cycle and mitochondrial membrane potential change of SKVO-3 cells were tested.The effects of SI-4650 on apoptosis,migration and invasion were investigated.The effects of SI-4650 on Bax,Bcl-2,Caspase3,E-cadherin,N-cadherin,Vimentin,matrix metalloproteinase 2(MMP2)and MMP 9 expression levels in SKVO-3 cells were detected.Results:Comparison between blank control group and experimental groups,SI-4650 could improve the content of SI-4650 in SKVO-3 cells.SI-4650 could inhibit the activity of SMO(P<0.01),reduce the ROS(P<0.01)and polyamine content in SKVO-3 cells(P<0.01).Treatment of SKVO-3 cells with SI-4650 inhibited the proliferation(the inhibition rate was 32.27%and 47.31%in experimental groups),caused S-phase cell cycle arrest(P<0.01)and induced apoptosis(P<0.01).The expressions of Bax and c-Caspase3 in SKVO-3 cells were increased(P<0.01),the content of Bcl-2 was decreased(P<0.01),and the mitochondrial membrane potential was decreased(P<0.01),and the number of apoptotic cells was increased(31.41%and 43.51%in experimental groups).At the same time,SI-4650 could change the expression levels of EMT-related factors,increased the expression level of E-cad,decreased the expression levels of N-cad,Vimentin,MMP-2 and MMP-9,and inhibited the migration and invasion of SKVO-3 cells.Conclusion:SI-4650 can effectively inhibit proliferation,invasion and metastasis of human ovarian cancer SKVO-3 cells,and the mechanism may be related to its ability to depress the activity of
作者
杨建林
田家俊
张浩
陈倩影
吕亚丰
曹春雨
YANG Jian-lin;TIAN Jia-jun;ZHANG Hao;CHEN Qiao-ying;LYU Ya-feng;CAO Chun-yu(Medical College of China Three Gorges University,Hubei Key Laboratory of Tumor Microenvironment and Immunotherapy,Yichang 443002,China)
出处
《中国应用生理学杂志》
CAS
CSCD
北大核心
2022年第2期175-180,192,共7页
Chinese Journal of Applied Physiology
基金
国家自然科学基金资助项目(81372265,30772590)
湖北省卫生健康科研基金(WJ2019H532)肿瘤微环境与免疫治疗湖北省重点实验室开放基金项目(2019KZL01,2016KZL04)。
关键词
精胺氧化酶抑制剂
人卵巢癌细胞
细胞培养
细胞增殖
凋亡
侵袭迁移
inhibitor of spermine oxidase
human ovarian cancer cell
cell culture
cell proliferation
apoptosis
migration and invasion
