光动力治疗对前列腺癌细胞膜蛋白影响及机制探讨

Effect and mechanism of photodynamic therapy on prostate cancer cell membrane protein

目的 近年来光动力治疗前列腺癌逐渐兴起,其疗效评价尚未统一。本研究旨在探讨光动力治疗后前列腺癌细胞膜蛋白变化机制及意义,为后续临床研究提供参考。方法 培养PC-3细胞分为A。B两组,MALDI-TOF/MSMS方法进行蛋白提取,蛋白质身份确定由美国国立生物技术信息中心(National Center for Biotechnology Information,NCBI)人类蛋白质数据库中检索,采用两相电泳联合串联质谱蛋白质组学方法分析并统计学分析电泳差异,rhodamine-123染色标记光动力处理之后线粒体膜通透性的改变,统计学分析荧光强度改变,对光动力治疗促使前列腺癌细胞死亡的作用靶点进行分析。结果 光动力治疗干预后,对照组与光处理组蛋白质总量及条带分布近似,线粒体60×10^3 热感蛋白(mitochondrial 60×10^3 heat shock protein,HSP60),Entrez Gene ID:31542947,PI 5.7,MW:61016.4,Protein Score:354,Protein Score C.I.%:100。电压依赖性的阴离子通道(voltage-dependent anion channel,VDAC),Entrez Gene ID:340201,PI 7.49,MW:31574.6,Protein Score:178,Protein Score C.I.%:100,蛋白电泳点差异有统计学意义,F=2.564,P=0.021;光动力处理后细胞rhodamine-123荧光强度由55.13降低到22.47,F=4.372,P=0.009,表明线粒体膜电位降低。细胞膜通透性在光动力作用后增加。结论 光动力治疗引发线粒体膜通透性增强,线粒体中与去极化效应相关的蛋白质表达上调,使线粒体的损伤不可逆转,最终导致癌细胞死亡,有可能成为光治疗疗效的指标。

OBJECTIVE In recent years,photodynamic therapy for prostate cancer has emerged,but the evaluation of its efficacy is not uniform.This study aimed to explore the mechanism and significance of changes in membrane proteins of prostate cancer cells after photodynamic therapy,and provide a reference for follow-up clinical research.METHODS Cultured PC-3 cells were divided into two groups:A and B,and MALDI-TOF/MSMS method was used for protein extraction,which was identified from the NCBI human protein database.Then used two phase electrophoresis combined with tandem mass spectrometry to analyze electrophoretic differences.Rhodamine-123 staining marked the change of mitochondrial membrane permeability after photodynamic treatment,and the change of fluorescence intensity was statistically analyzed.The role of photodynamic therapy in promoting the death of prostate cancer cells was analyzed.RESULTS After photodynamic therapy,the total protein content and band distribution of the control group were similar to that of the photodynamic treatment group.Mitochondrial heat shock protein(HSP60),Entrez Gene ID:31542947,PI5.7,MW:61016.4,Protein Score:354,Protein Score C.I.%:100.Voltage-dependent anion channel(VDAC),Entrez Gene ID:340201,PI 7.49,MW:31574.6,Protein Score:178,Protein Score C.I.%:100,the electrophoretic point of protein was significantly different,F=2.564,P=0.021;the fluorescence intensity of rhodamine-123 decreased from 55.13 to 22.47 after photodynamic treatment,F=4.372,P=0.009,indicating a decrease in mitochondrial membrane potential.The permeability of cell membrane was increased after photodynamic therapy.CONCLUSIONS Photodynamic therapy enhances changes in mitochondrial membrane permeability,leading to the eventual death of cancer cells.The expression of proteins related to depolarization effect in mitochondria is up-regulated,which makes mitochondrial damage irreversible,that may become an indicator of the efficacy of phototherapy.