摘要
为研究沙棘籽渣黄酮类化合物(FHR)对人乳腺癌细胞Bcap-37的生长抑制和凋亡的诱导作用,探讨其抗癌作用的机制,本项研究应用CCK-8法检测FHR对Bcap-37的生长抑制效应,流式细胞术(FCM)分析细胞周期及凋亡率,透射电镜观察细胞凋亡的超微结构表征,半定量RT-PCR方法和Westernblot方法检测与细胞周期及凋亡调控相关基因的转录和表达的变化。结果表明,FHR可明显抑制Bcap-37的增殖,并呈剂量依赖性;FCM分析发现在FHR1000μg/ml作用24h后能阻断Bcap-37细胞于G2/M期,并伴有少量的亚G1峰出现;TEM观察Bcap-37细胞经FHR作用后具有典型的凋亡形态学特征;半定量RT-PCR结果显示,经1000μg/mlFHR作用24h后,PCNA、Bcl-2转录水平分别下调为对照组的0.26倍和0.66倍,IGFBP4转录水平上调为对照组的1.13倍。Westernblot结果显示p53蛋白表达显著上调,Bcl-2蛋白表达显著下调。本项研究结果提示FHR可能是通过下调PCNA的转录使细胞的增殖被阻断在G2/M期,并通过p53蛋白上调和Bcl-2蛋白的下调诱导Bcap-37细胞凋亡。
To investigate the effects of flavonoids from Hippophae rhamnoides (FHR)-indueed growth inhibition and apoptosis in human breast cancer cell line Bcap-37, the antiproliferative effect of FHR against Bcap-37 cells was tested by CCK-8 method, cell apoptosis percentage and the cell cycle phase distribution of Bcap-37 cells were measured by flow cytometric assay, apoptotic phenotype of Bcap-37 was observed by transmission electron microscopy, the variation of transcription and expression of cell cycle and apoptosis related genes was determined by semi-quantitative RT-PCR and Western blot. The results showed that a dose-dependent proliferation inhibition was demonstrated in Bcap-37 cells, and a G2/M cell cycle arrest was induced by FHR. The characteristic morphological changes of apoptosis in Bcap-37 cells were observed after treatment with FHR for 24 h. The transeriptional level of PCNA and Bcl-2 were down-regulated respectively to 26% and 66% of its mRNA of the control group, whereas, the mRNA transeriptional level of IGFBP4 was up-regulated to 1.13 times of that of the control group. The expression level of p53 protein was up-regulated and Bcl-2 protein was down-regulated. These results indicate that the growth inhibition effect by means of dose-dependent manner of FHR on breast cancer cells Bcap-37 seems to be down-regulated by PCNA which resulted in a G2/M cell cycle arrest. FHR induced apoptosis of Bcap-37 might be due to the up-regulation of p53 and down-regulation of Bcl-2.
出处
《现代免疫学》
CAS
CSCD
北大核心
2005年第2期98-101,107,共5页
Current Immunology
基金
上海市科技发展基金资助项目(014358009)"十五"
"211工程"重点学科建设子项目
