CAJ | 학술논문

目的探讨天花粉蛋白对人乳腺癌耐药细胞株MCF-7/ADR及干细胞标志物ALDH1蛋白表达的影响.方法主要采用MTT法检测不同浓度的天花粉蛋白(TCS)对MCF-7/ADR细胞增殖的抑制作用;流式细胞术(FCM)检测MCF-7/ADR细胞的凋亡情况;四唑氮盐(MTT)比色试验、Western-Blot印迹法观察天花粉蛋白对乳腺癌多药耐药细胞株MCF-7/ADR的耐药逆转作用.结果 MTT分析结果显示,TCS在20~100mg/L浓度范围内有明显的抑制作用,存在剂量-效应和时间-效应关系.流式细胞术检测结果显示:20,40,80mg/L TCS作用48h后,MCF-7/ADR细胞凋亡率分别为12.56%±0.70%,18.34%±1.01%,38.92%±1.79%,细胞凋亡率随药物浓度增加而增加,各实验组之间比较有显著性差异(P<0.05).(MTT)比色试验、Western-Blot印迹法结果发现:MCF-7中未测出ALDH1蛋白,在MCF-7/ADR空白对照和TCS 20mg/L作用2d后检测出ALDH1蛋白表达开始下降,MCF-7/ADR在TCS 20mg/L作用3d,4d后ALDH1蛋白表达明显下降,MCF-7/ADR在TCS 20mg/L作用5d后未检测到ALDH1蛋白的表达.结论天花粉蛋白(TCS)具有逆转乳腺癌细胞株MCF-7/ADR耐药性的作用,可明显下调ALDH1蛋白的表达.天花粉蛋白(TCS)可能通过下调ALDH1蛋白的表达,抑制乳腺癌干细胞的富集,来诱导MCF-7/ADR细胞凋亡,抑制MCF-7/ADR细胞的增殖.
목적 탐토천화분단백대인유선암내약세포주MCF-7/ADR급간세포표지물ALDH1단백표체적영향.방법 주요채용MTT법검측불동농도적천화분단백(TCS)대MCF-7/ADR세포증식적억제작용;류식세포술(FCM)검측MCF-7/ADR세포적조망정황;사서담염(MTT)비색시험、Western-Blot인적법관찰천화분단백대유선암다약내약세포주MCF-7/ADR적내약역전작용.결과 MTT분석결과현시,TCS재20~100mg/L농도범위내유명현적억제작용,존재제량-효응화시간-효응관계.류식세포술검측결과현시:20,40,80mg/L TCS작용48h후,MCF-7/ADR세포조망솔분별위12.56%±0.70%,18.34%±1.01%,38.92%±1.79%,세포조망솔수약물농도증가이증가,각실험조지간비교유현저성차이(P<0.05).(MTT)비색시험、Western-Blot인적법결과발현:MCF-7중미측출ALDH1단백,재MCF-7/ADR공백대조화TCS 20mg/L작용2d후검측출ALDH1단백표체개시하강,MCF-7/ADR재TCS 20mg/L작용3d,4d후ALDH1단백표체명현하강,MCF-7/ADR재TCS 20mg/L작용5d후미검측도ALDH1단백적표체.결론 천화분단백(TCS)구유역전유선암세포주MCF-7/ADR내약성적작용,가명현하조ALDH1단백적표체.천화분단백(TCS)가능통과하조ALDH1단백적표체,억제유선암간세포적부집,래유도MCF-7/ADR세포조망,억제MCF-7/ADR세포적증식.
@@@@ Objective To study the effects of trichosanthin on MCF-7/ADR resistant cell line of human breast cancer and stem cell markers ALDH1 protein expression.Methods MTT was mainly used to detect proliferation inhibition of MCF-7/ADR cells for different concentrations of TCS.FCM was used to detect apoptosis in MCF-7/ADR cells.MTT colorimetric test and Western-Blot were used to observe trichosanthin resistance reversal effect of multidrug resistance in breast cancer cell line MCF -7/ADR.Results MTT assay showed that within the concentration of 20~100mg/L,TCS had significantly inhibition effects in a dose-response and time-effect relationship.The results of FCM showed that after 48h of 20,40 and 80mg/L TCS action,the apoptosis rate of MCF-7/ADR were 12.56%±0.70%,18.34%±1.01% and 38.92%±1.79%respectively,the rate of apoptosis with the drug concentration increased among the experimental group significantly .MCF-7/ADR cells were treated with different concentrations of TCS after 48h,the occurrence of S-phase was arrested,compared with the control group,the difference was statistically significant .MTT colorimetric test and the Western-Blot blot results found:ALDH1 protein was not detec-ted in MCF-7,but in the MCF-7/ADR blank control and TCS 20mg/L,the expression of ALDH1 protein declined after 2d;in the TCS 20 mg/L group of 3d and 4d the expression of ALDH1 protein in MCF-7/ADR decreased,while it was not detected after 5d.Conclusion TCS has the role of reversing MCF-7/ADR resistance,significantly lowering the expression of ALDH 1 protein,possibly inhibiting the enrichment of breast cancer stem cells,inducing MCF-7/ADR cells apoptosis,and inhibiting the proliferation of MCF-7/ADR cells.