着色性干皮病D组蛋白对胰腺癌PANC-1细胞紫杉醇敏感性的影响  

Effects of XPD on paclitaxel senstivity of pancreatic cancer cell line PANC-1

作　　者：简捷[1] 刘利珍 邓峰[1] 黄缘[3] 秦健斌[1] 李双[1] 

机构地区：[1]南昌大学第三附属医院消化内科,南昌医学硕士330008 [2]上海市嘉定区中心医院肿瘤科,上海201800 [3]南昌大学第二附属医院消化内科,南昌330006

出　　处：《医学研究生学报》2016年第12期1286-1290,共5页Journal of Medical Postgraduates

摘　　要：目的研究表明化疗药物耐药和与消化道肿瘤相关的一系列基因分子突变密切相关。文章研究野生型人剪切修复基因XPD转染人胰腺癌PANC-1细胞后对结合型紫杉醇敏感性的影响。方法将空载质粒p EGFP-N_2及重组质粒p EGFP-N_2/XPD通过瞬时转染于人胰腺癌PANC-1细胞。设正常对照组(不作任何处理)、空载质粒转染组(空载质粒p EGFPN_2转染)、重组质粒XPD转染组(重组质粒p EGFP-N_2/XPD转染)。使用RT-PCR和Western blot方法检验转染后XPD mRNA及蛋白表达;MTT实验检测结合型紫杉醇对XPD转染后PANC-1细胞增殖活性的变化;流式细胞仪检测XPD转染后PANC-1细胞在结合型紫杉醇作用下凋亡反应的差异。结果重组质粒XPD转染组中XPD mRNA表达量(9.572±0.345)和蛋白表达量(1.055±0.012)较正常对照组(1.125±0.043,0.302±0.002)及空载质粒转染组(1.273±0.057,0.378±0.004)明显升高(P<0.01);正常对照组与空载质粒转染组XPD mRNA和蛋白表达量差异无统计学意义(P>0.05)。MTT结果显示不同浓度结合型紫杉醇作用PANC-1细胞24 h后,重组质粒XPD转染组的A值均明显低于正常对照组、空载质粒转染组,差异均有统计学意义(P<0.05);正常对照组与空载质粒转染组差异无统计学意义(P>0.05)。流式细胞仪检测结果显示不同浓度结合型紫杉醇作用PANC-1细胞24 h后,重组质粒XPD转染组细胞凋亡率均明显高于正常对照组、空载质粒转染组,差异均有统计学意义(P<0.01);正常对照组与空载质粒转染组差异无统计学意义(P>0.05)。结论通过XPD转染PANC-1细胞可明显增强PANC-1细胞对结合型紫杉醇的化疗敏感性。Objective Studies have shown that chemotherapy drug resistance is closely associated with a series of gene muta-tion related to digestive tract tumor. The aim of this study was to explore the effects of transfected Xeroderma Pigmentosum D （ XPD） on paclitaxel sensitivity of human pancreatic cancer cell line PANC-1. Methods Blank plasmid pEGFP-N2and recombinant plasmid pEGFP-N2/XPD were transiently transfected into PANC-1, respectively. They were divided into three groups ： Normal control group （no treatment）,Blank plasmid pEGFP-N2 transfected group （transfected with blank plasmid pEGFP-N2） and Recombinant plasmid pEGFP-N2/XPD transfected group （transfected with recombinant plasmid pEGFP-N2/XPD）. Reverse transcriptase PCR and Western blot were used to detect the expressions of XPD mRNA and protein, respectively. The sensitivity of PANC-1 cells to paclitaxel after XPD transfection was assessed by MTT assay. Flow cytometry was used to examine PANC-1 cells apoptosis exposed to paclitaxel after XPD transfection. Results Compared with Normal control group （1.125±0.043,0 .3 0 2 ± 0 .002） and Blank plasmid pEGFP-N2tmns-fected group （1.273±0.057, 0.378±0 .00 4） ,the expression of XPD mRNA （9 .572±0.345） and protein （1.055±0.012） in Recombi-nant plasmid pEGFP-N2/XPD transfected group was significantly increased （P〈0.01）. MTT assay showed the proliferation of PANC-1 cells exposed to paclitaxel for 24 hours was significantly inhibited after transfection of XPD compared with Normal control group and Blank plasmid pEGFP-N2 transfected group（ P〈0.05） .Normal control group and Blank plasmid pEGFP-N2 transfected group had no statistically difference（P〉0.05） .Flow cytometry showed that the apoptosis of PANC-1 cells exposed to paclitaxel for 24 hours was sig-nificantly increased after XPD transfection（ P〈0.01） .The apoptosis exposed to paclitaxel in Normal control group and Blank plasmid pEGFP-N2 transfected group

关 键 词：着色性干皮病D组蛋白 人胰腺癌细胞株PANC-1 紫杉醇 增殖 凋亡 

分 类 号：R735.9[医药卫生—肿瘤]