摘要
该文初步探讨了环吡酮胺(ciclopirox olamine, CPX)抑制人胶质瘤细胞SHG44生长的作用和机制。该研究用梯度浓度的CPX处理人胶质瘤细胞SHG44后,通过MTT实验检测药物半抑制浓度IC50和细胞增殖能力;应用平板克隆实验检测细胞克隆形成能力;Transwell小室和细胞划痕实验检测细胞侵袭和迁移能力;流式细胞术检测细胞内活性氧(reactive oxygen species, ROS)、线粒体内ROS以及细胞凋亡的变化;Seahorse XF96 Flux analysis分析仪检测细胞耗氧率(oxygen consumption rate, OCR);实时荧光定量PCR(q PCR)和Western blot技术检测细胞内m RNA和蛋白的变化。结果表明:SHG44对CPX药物敏感;CPX能使STAT3在m RNA转录水平和蛋白表达水平下降,抑制SHG44的迁移和侵袭,同时,促进线粒体ROS的累积,最终破坏线粒体功能,抑制细胞生长并促进细胞凋亡。该研究结果提示,CPX具有一定的抗胶质瘤细胞SHG44生长的作用,有望成为一种治疗胶质瘤的药物。
The aim of this study was to investigate the effects and molecular mechanism of ciclopirox olamine(CPX)inhibiting the growth of human glioma cell SHG44.In this study,human glioma cell line SHG44 was treated with a gradient concentration of CPX,then the half maximal inhibitory concentration(IC50)and cell proliferation ability of the CPX treated SHG44 cells were determined by MTT assay.In addition,the clone-forming ability was detected to confirm the effect of CPX on SHG44 cell growth.Moreover,the transwell cell migration and invasion assays were performed to study the effects of CPX on the capacity of cell motility and invasiveness.Intracellular reactive oxygen species(ROS),mitochondrial ROS,and apoptosis were detected by flow cytometry.Seahorse XF96 Flux analyzer was used to measure the oxygen consumption rate(OCR),and Real-time PCR(qPCR)as well as Western blot were applied to detect the mRNA and protein levels.Our results showed that SHG44 cells were sensitive to CPX treatment.CPX could reduce both the mRNA and protein expression levels of STAT3,and thus inhibiting the migration and invasion of SHG44 cells.Moreover,CPX treatment significantly enhances the accumulation of mitochondrial ROS,which leads to the impairement of mitochondrial function,and thus inhibiting cell growth and promoting apoptosis.Collectively,our findings demonstrated that CPX could inhibit the growth of SHG44 cells,which may be developed as a drug for the treatment of glioma.
作者
韩胜男
龚世伟
费明明
上官福根
吕斌
Han Shengnan;Gong Shiwei;Fei Mingming;Shangguan Fugen;LüBin(School of Laboratory Medicine and Life Sciences,Wenzhou Medical University,Wenzhou,Zhejiang325035,China;Zhejiang Provincial Key Laboratory of Medical Genetics,Wenzhou325035,China)
出处
《中国细胞生物学学报》
CAS
CSCD
2019年第5期933-941,共9页
Chinese Journal of Cell Biology
基金
国家自然科学基金(批准号:31771534、31570772)资助的课题~~
