CAJ | 학술논문

目的研究和探讨力达霉素对碱性成纤维细胞生长因子(bFGF)诱导的癌细胞中信号转导的抑制作用.方法MTT法检测LDM和阿霉素(ADR)对3种人癌细胞系的增殖抑制作用.受体结合实验检测LDM对bFGF与其受体结合的作用.Western blotting检测bFGF受体复合物的形成,流式细胞测定细胞内Ca2+的流动,免疫印迹法测定bFGF所诱导的3种PKC亚型的活性.结果MTT法实验结果,LDM在体外对3种人癌细胞均显示出强烈的增殖抑制作用.按IC50值进行比较表明,LDM的活性比ADR强1 000倍以上.LDM抑制[125I]-bFGF对大鼠肺细胞膜受体的IC50为2.0×10-4nmol·L-1.LDM能阻碍bFGF与其受体复合物的形成,LDM(10 nmol·L-1)预孵育2 h可阻止bFGF诱导的细胞内Ca2+效应.免疫印迹法证明,LDM可抑制bFGF诱导的癌细胞的3种PKC亚型的活性.结论力达霉素抗肿瘤作用的机制之一可能是通过阻断bFGF受体的信号转导途径.
목적연구화탐토력체매소대감성성섬유세포생장인자(bFGF)유도적암세포중신호전도적억제작용.방법MTT법검측LDM화아매소(ADR)대3충인암세포계적증식억제작용.수체결합실험검측LDM대bFGF여기수체결합적작용.Western blotting검측bFGF수체복합물적형성,류식세포측정세포내Ca2+적류동,면역인적법측정bFGF소유도적3충PKC아형적활성.결과MTT법실험결과,LDM재체외대3충인암세포균현시출강렬적증식억제작용.안IC50치진행비교표명,LDM적활성비ADR강1 000배이상.LDM억제[125I]-bFGF대대서폐세포막수체적IC50위2.0×10-4nmol·L-1.LDM능조애bFGF여기수체복합물적형성,LDM(10 nmol·L-1)예부육2 h가조지bFGF유도적세포내Ca2+효응.면역인적법증명,LDM가억제bFGF유도적암세포적3충PKC아형적활성.결론력체매소항종류작용적궤제지일가능시통과조단bFGF수체적신호전도도경.
Aim To study the mechanism of inhibition of basic fibroblast growth factor (bFGF) related signal transduction by lidamycin in cancer cells. Methods MTT assay was used to determine the growth inhibitory effect of lidamycin (LDM) and adriamycin (ADR) in several cancer cell lines. The inhibition of bFGF bound to its receptor by LDM was measured with [ 125I ]-bFGF binding assay.Intracellular Ca2+ stimulated by bFGF was measured by Fura-3. The formation of bFGF- receptor immune complex and the inhibitory effect of LDM on the activity of PKC isoenzymes induced by bFGF in cancer cells were identified by Western blotting analysis. Results LDM displayed extremely potent growth inhibitory effect on several cancer cell lines in a dose-dependent manner. A comparison of the IC50 values showed that the effect of LDM was 1 000-fold more potent than that of ADR. LDM blocked the specific by anti-FGFR specific antibody, LDM inhibited the formation of bFGF-receptor immune complex. bFGF demonstrated that LDM inhibited the activity of PKC isoenzymes in cancer cells stimulated with bFGF.Conclusion The blocking of bFGF receptors in the signal transduction pathway may be involved in the effect of LDM on cancer cells.