摘要
目的研究人喉咽癌细胞株FaDu与其耐药株FaDu/T在生物学特性方面的异同及内在联系。方法以喉咽癌FaDu细胞为亲本,通过紫杉醇(Taxol)递增法筛选出其耐药细胞株FaDu/T。MTT法检测其耐药性,流式细胞术检测其周期,吖啶橙和Hoechest 33342/PI染色法检测细胞凋亡情况;RT-PCR和Western blot检测多药耐药、凋亡相关基因。结果与FaDu细胞相比,FaDu/T细胞具有以下特点:①生长缓慢;②多药耐药性增强;③处于G0/G1期较多,而G2/M和S期较少,凋亡比例无明显差别;④耐药基因ABCB1表达增加;⑤抗凋亡蛋白Bcl-2表达增加,凋亡蛋白Bax、Caspase-3表达下降。结论 FaDu/T细胞中耐药及抗凋亡蛋白ABCB1、Bcl-2的高表达是肿瘤细胞化疗抵抗的关键原因,肿瘤细胞各生物学特性的内在联系有待进一步研究。
OBJECTIVE To investigate the different or similar characteristics between the hypopharyngeal carcinoma cell line（FaDu cells）and the multidrug resistance（MDR）cell lines（FaDu/T）induced by Taxol.METHODS Human FaDu cells were exposed in stepwise escalating concentration of Taxol until the resistant cell line was formed.The drug resistance was investigated by methyl-thiazolyl-tetrazolium（MTT）assay.The cell apoptosis was measured by staining cells with acridine orange（AO）,Ho.33342 and PI staining.Meanwhile,the cell cycle distribution was quantified through flow cytometry.Reverse Transcription Polymerase Chain Reaction（RT-PCR）was conducted to detect mRNA levels of the drug resistance genes ABCB1 and apoptosis related genes Bax,Bcl-2,Caspase-3.The proteins levels were detected through Western blot.RESULTS Compared with parents FaDu cells,the drug resistance cells of FaDu/T had the biological characteristics as follows： ①They had a lower growth velocity;②Their mutidrug resistance ability was increased;③The percentage of cells in G0/G1 phases was increased while that in G2/M and S phase decreased,but the proportion of apoptosis cells had no prominent difference;④The expression of ABCB1 gene increased significantly;⑤The anti-apoptotic protein Bcl-2 increased prominently,while the apoptotic protein,Caspase-3 and Bax,decreased.CONCLUSION The high expression of ABCB1,Bcl-2 may be the critical reason of the chemotherapy resistance in FaDu/T;but the internal connection among various biological characteristics of tumor cells is worth to be further investigeated.
出处
《中国耳鼻咽喉头颈外科》
北大核心
2011年第12期658-662,共5页
Chinese Archives of Otolaryngology-Head and Neck Surgery
关键词
下咽肿瘤
细胞
培养的
多药耐药相关蛋白质类
细胞凋亡
流式细胞术
Hypopharyngeal Neoplasms
Cells
Cultured
Multidrug Resistance-Associated Proteins
Apoptosis
Flow Cytometry
作者简介
第一作者简介及通讯：马聚珂，男，河南人，在读医学硕士，主要从事头颈肿瘤的研究。Email：majuke99@163．com
通讯作者：徐伟（Email：xwhns@yahoo．com．cn）
