芳香烃受体(AhR)在乳腺癌中的表达及其与阿霉素化疗耐药的相关性  

Aryl hydrocarbon receptor(AhR)expression in breast cancer tissues and its association with adriamycin chemotherapy resistance

作　　者：李正东[1] 杨新伟[2] 成小林[1] 庄志刚[1] 童晓文[3] 

机构地区：[1]同济大学附属第一妇婴保健院乳腺外科,上海200040 [2]上海中医药大学附属市中医医院中医外科,上海200071 [3]同济大学附属同济医院妇产科上海200065

出　　处：《复旦学报（医学版）》2014年第1期93-97,共5页Fudan University Journal of Medical Sciences

基　　金：上海市科委资助课题(134119a7900)~~

摘　　要：目的观察芳香烃受体(aryl hydrocarbon receptor,AhR)在乳腺癌组织中的表达,并探讨乳腺癌细胞AhR表达与阿霉素化疗耐药的关系。方法应用免疫组化染色法观察AhR在50例乳腺癌标本中的表达,其中淋巴结转移癌40例,正常乳腺组织10例。采用AhR-siRNA表达载体和脂质体法瞬时转染基因沉默AhR高表达的乳腺癌细胞株MCF-7/ADR;RT-PCR和Western bolt法检测转染后AhR mRNA及蛋白的表达;MTT法检测细胞增殖活性及转染前后乳腺癌细胞对阿霉素敏感性的变化。结果 AhR在乳腺癌组织表达率为82.0%(46/50)、在乳腺癌转移淋巴结中表达率为92.5%(37/40),而在正常乳腺组织中10%(1/10)有表达。乳腺癌及乳腺癌转移淋巴结中AhR的表达均显著高于正常乳腺组织(P<0.05)。基因沉默AhR高表达的乳腺癌细胞株MCF-7/ADR后24h,PCR和免疫印迹结果均显示乳腺癌细胞株AhR基因及蛋白表达水平降低。MTT显示AhR基因沉默对乳腺癌MCF-7/ADR细胞的增殖活性有显著的抑制作用,48h细胞增殖抑制率可达52%。AhR沉默后乳腺癌细胞对阿霉素的半数有效浓度(IC50)由(18.2±0.9)μmol/L降低至(8.4±1.1)μmol/L(P<0.05)。结论 siAhR能够有效抑制AhR基因的表达,降低乳腺癌细胞的增殖能力。AhR对阿霉素耐药过程发挥作用,沉默AhR表达能一定程度上逆转阿霉素耐药现象。Objective To investigate both the expression of aryl hydrocarbon receptor （AhR） in breast cancer tissues and its association with adriamycin chemotherapy resistance. Methods AhR expression was observed by immunohistochemical staining in 50 specimens of breast cancer containing 40 cases of metastasis lymph nodes and 10 cases of normal breast tissues. Breast cancer cell line MCF- 7/ADR with AhR high expression was transfected by the AhR-siRNA liposome vector. The expression of AhR mRNA and protein was detected by RT-PCR and Western blot after transfection.Adriamycin sensitivity of breast cancer cells before and after transfection was determined by proliferation test,MTT. Results AhR in immunohistochemical staining was expressed in 82% （46/ 50） breast cancer tissue,in 92.5 （37/40） metastasis lymph nodes,and in 10% （1/10） normal breast tissue. AhR expression in normal breast tissues was significantly lower than that in breast cancers and metastasis lymph nodes （P〈0.05）. PCR and Western blot results showed that expression of AhR gene and protein in breast cancer cell line MCF-7/ADR was decreased 24 hours later after AhR gene silencied by liposome transfection. The proliferation of breast cancer cell MCF-7/ADR was significantly inhibited after AhR gene was silenced. MTT showed the inhibition rate of ceil proliferation was up to 52 % at 48 hours. Median effective concentration （ICs0） to adriamycin was decreased from （18.2 ± 0.9） u mol/L to （8.4 ± 1. 1） u mol/L （P%0.05） before and after AhR-silenced in breast cancer cells. Conclusions AhR-siRNA can effectively inhibit the expression of AhR gene and reduce the proliferation of breast cancer cell. AhR plays a role in adriamycin resistant process, so that AhR silencing can partly reverse adriamycin resistant in breast cancer.

关 键 词：乳腺癌 RNA干扰 化疗耐药 阿霉素 

分 类 号：R735.9[医药卫生—肿瘤]