TUBB3-ASODN对食管鳞癌细胞Ec9706/P-1耐药性的影响及机制探讨  

Effect and mechanism of β-tubulin Ⅲ antisense oligonucleotides on paclitaxel-resistant of human esophageal carcinoma cell line Ec9706/P-1

作　　者：卢红[1] 樊青霞[2] 王瑞林[2] 

机构地区：[1]河南大学淮河医院,河南开封475001 [2]郑州大学第一附属医院

出　　处：《山东医药》2010年第18期10-11,共2页Shandong Medical Journal

摘　　要：目的观察β-微管蛋白Ⅲ(TUBB3)反义寡核苷酸(ASODN)对食管鳞癌紫杉醇耐药细胞Ec9706/P-1耐药性的影响,并探讨其机制。方法将EC9706/P-1细胞分为A、B、C组,A组不作处理,B、C组分别加入无义寡核苷酸(NSODN)、TUBB3-ASODN转染24 h,加入100、10、1、0.1、0.001μg/ml的紫杉醇培养24 h。采用MTT法测算各组紫杉醇半数抑制浓度(IC50)及细胞耐药指数(RI),采用流式细胞仪检测细胞周期,采用RT-PCR和免疫组化法检测TUBB3 mRNA及其蛋白。结果C组紫彬醇IC50为(7.78±1.15)μg/ml、RI为3.37,B组分别为(16.56±1.80)μg/ml、7.17,A组分别为(15.40±2.06)μg/ml、6.67,C组与A、B组相比,P均<0.05。C组G0+G1期细胞百分比为78.7%±2.3%,S期为14.3%±2.1%,G2+M期为7.0%±0.9%;B组分别为87.3%±1.2%、6.9%±0.4%、5.8%±0.8%,A组分别为85.9%±1.1%、7.5%±0.5%、6.7%±0.9%,C组G0+G1期、S期细胞百分比与A、B组比较,P均<0.05。C组TUBB3蛋白阳性率为33.68%±8.34%、TUBB3 mRNA为1.33±0.08,B组分别为62.96%±10.49%、1.59±0.06,A组分别为64.47%±11.75%、1.60±0.10,C组与A、B组相比,P均<0.05。结论TUBB3-ASOND可增加Ec9706/P-1对紫杉醇的敏感性,TUBB3升高可能是食管鳞癌对紫杉醇产生耐药的机制之一。Objective To observe the effect of β-tubulinⅢ（TUBB3）antisense oligonucleotide（ASODN）on paclitaxel-resistant of human esophageal carcinoma cell line Ec9706/P-1,and explore its mechanism.Methods Human esophageal carcinoma cell line Ec9706/P-1 cell were divided into group A,B,C.Cells in group B and C were cultured with NSODN for 24 h.100、10、1、0.1、0.001 μg/ml paclitaxel were mix in group A,B,C for 24 h.Flow cytometry（FCM）was performed to analyze cell cycle.The proliferation inhibitory rate and drug resistance index（RI）were evaluated by MTT assay.TUBB3 was determined by immunohistochemistry.TUBB3 mRNA expression was detected by RT-PCR.Results The 50% inhibiting concentration（IC50）and RI in group C was（7.78±1.15）μg/ml and 3.37.These were（16.56±1.80）μg/ml and 7.17 in group B,（15.40±2.06）μg/ml and 6.67 in group A.The IC50 and RI in group C decreased compared with other groups（P0.05）.The expression of TUBB3 mRNA（1.33±0.08）and protein（33.68%±8.34%）were obviously decreased（P0.05）respectively in the group of C.Conclusions TUBB3 ASODN can increase the sensitivity of the Ec0706/P-1 to paclitaxel significantly.Overexpression of TUBB3 is one of factors responsible for the development of paclitaxel-resistance.

关 键 词：β-微管蛋白Ⅲ 反义寡核苷酸 食管肿瘤 鳞状细胞癌 紫杉醇 细胞耐药 

分 类 号：R735.1[医药卫生—肿瘤]