天蚕素A-爪蟾素2杂合肽P18对MDA-MB-435s乳腺癌细胞的作用研究

The Effect of Cecropin A-Magainin2 Hybrid Antimicrobial Peptide P18 on the MDA-MB-435s Cell Line

目的研究天蚕素A-爪蟾素2杂合肽P18对体外培养的人类乳腺癌细胞株MDA-MB-435s的活性作用。方法采用MTT法检测P18对体外培养的MDA-MB-435s细胞增殖的影响;使用Live/Dead Viability/Cytotoxicity试剂盒对细胞进行染色,并在荧光显微镜下观察细胞的存活状态;通过DiBAC4(3)染色,以荧光分光光度计检测给肽时细胞膜的膜电位变化;在透射电子显微镜下观察P18作用24h后细胞超微结构的变化。结果MTT试验表明P18对MDA-MB-435s细胞的增殖具有浓度依赖性的抑制作用;荧光显微镜下可观察到P18可以使MDA-MB-435s细胞膜破损,细胞死亡;P18作用时,荧光分光光度计检测到MDA-MB-435s的细胞膜发生了去极化现象;透射电子显微镜下可见大量细胞崩解坏死。结论杂合肽P18能够引起MDA-MB-435s细胞膜的通透性改变而导致细胞坏死。

Objective To study the effect of cecropin A-magainin2 hybrid antimicrobial peptide P18 on the human breast cancer cell line MDA-MB-435s. Methods Cultured MDA-MB-435s cells were treated by P18 and cell proliferation was assessed by MTr assay; the state of the cells was observed with a Live/Dead Viability/Cytotoxicity kit by fluorescence microscopy ; the change of cell membrane potential was monitored with the existence of DiBAC4 （ 3 ） by spectrofluorometer ; and the uhrastructure of MDA-MB-435s cells were observed by transmission electron microscopy after 24 hour P18 treatment. Results MTT test shows that P18 can inhabit the proliferation of MDA-MB-435s cells in a dose dependent manner; dead cells with broken cell membranes can be seen under fluorescence microscopy; the depolarization process can be detected by speetrofluorometer; cell necrosis was observed by transmission electron microscopy. Conclusion P18 can enhance the membrane permeability of MDA-MB-435s cells and trigger cell necrosis.