mdr1基因在多药耐药相关的几种人胃癌细胞系的表达  

Expression of mdr1 gene in several human gastric cancer cell lines associated with multidrug resistance

作　　者：高福莲[1] 蔡新华[1] 马开颜[1] 乐晓萍[1] 张钦宪[1] 

机构地区：[1]郑州大学基础医学院分子细胞生物学研究中心河南省分子医学重点学科开放实验室,河南郑州450052

出　　处：《中国现代医学杂志》2006年第10期1490-1494,共5页China Journal of Modern Medicine

摘　　要：目的探讨mdr1在多药耐药相关的几种人胃癌细胞系中的转录、翻译、P-gp功能变动趋势。方法培养SGC7901/VCR(人胃癌多药耐药细胞亚系)、SGC7901和BGC823(人胃腺癌细胞系)于RPMI1640,用RT-PCR和原位杂交检测mdr1mRNA的表达,用免疫印迹和免疫组化检测P-gp的表达,用流式细胞仪检测阿霉素在细胞内的蓄积。结果半定量RT-PCR显示,SGC7901/VCRmdr1和β-actin吸收峰面积之比为5.63,SGC7901为0.61,BGC823为0.85。杂交信号呈紫蓝色颗粒,分布于胞质。免疫印迹显示SGC7901/VCRP-gp的表达最强,SGC7901最弱。P-gp阳性呈棕黄色颗粒,位于细胞膜上。SGC7901中,少数细胞P-gp表达极强。SGC7901/VCR平均光密度为(1.8310±0.8401),SGC7901为(0.3590±0.2512),BGC823为(0.6260±0.4996)(P<0.05)。SGC7901/VCR阿霉素特异荧光强度为(6.59±50.30),SGC7901为(35.88±14.55),BGC823为(27.44±7.06)(P<0.001)。结论在多药耐药相关的人胃癌细胞系SGC7901/VCR、SGC7901和BGC823中,mdr1基因均表达,P-gp具有药物外排功能,SGC7901/VCR的mdr1表达最强,SGC7901最弱,BGC823居中。[Objective] To explore alteration of mdrl transcription, translation and P-gp function in several human gastric cancer ceU lines associated with multidrug resistance. [Methods] SGC7901/VCR （the human gastric cancer multidrug resistance cell subline）, SGC7901 and BGC823（the human gastric adenocarcinoma cell line） were cultured in RPMI1640 respectively. The expression of mdrl mRNA was detected by RT-PCR and hybridization in situ. The P-gp expression was detected by immunoblotting and immunohistochemistry. The fluorescence intensity of intracellular adriamycin was assessed by flowcytometry. [Results] By the semiquantitative RT-PCR, the ratio of mdrl and β-actin was 5.63 in the SGC7901/VCR, 0.61 in the SGC7901 cells and 0.85 in the BGC823.The hybridization signals in violet-colored granules were dispersed in the cytoplasm. Based on the immunoblott/ng, the expression level of P-gp in the SGC7901/VCR cells was highest and that in the SGC7901 cells was lowest. The P-gp immunoreactivity （IR） in brownish-colored granules was located at the cell membrane region.The stronger P-gp-IR displayed only in a few SGC7901 cells. The averages of OD were （1.8310±0.8401）, （0.359±0.2512）, （0.6260±0.4996） in the SC, C7901/VCR, SGC7901, BGC823 cells respectively（P 〈0.05）. The values of adriamycin-specific fluorescence intensity were （6.59±50.30）, （35.88±14.55）, （27.44±7.06） in the SGC7901/VCR, SGC7901 and BGC823 cells respectively （P 〈0.001）. [Conclusions] The mdrl gene can be expressed in the human gastric cancer cen lines associated with multidmg resistance and the P-gp has the drag-exclusive function. Among these 3 cen lines, the mdrl expression level is the strongest in the SGC7901/VCR cens, weakest in the SGC7901 and moderate level in the BGC823 cells.

关 键 词：多药耐药基因1 P-糖蛋白 SGC7901/VCR细胞亚系 BGC823细胞系 SGC7901细胞系 

分 类 号：R735.2[医药卫生—肿瘤]