BCRP基因在非小细胞肺癌组织和非癌肺组织中的表达及意义

R734.2; 目的:进一步研究从人类乳腺癌细胞株(MCF-7/AdrVp)中克隆出的、对蒽环类抗癌药耐药的BCRP基因在非癌肺组织及非小细胞肺癌组织中的表达。方法:从术后即刻获得的正常肺组织和有活力的非小细胞肺癌组织新鲜标本中及时提取细胞总RNA,通过RT-PCR及PCR法获得并扩增BCRP基因的cDNA产物,再经凝胶电泳分离BCRP基因cDNA带,然后再通过转膜技术把这些BCRP基因cDNA产物转移到杂交膜上,用Southernblot杂交技术检测BCRP基因的表达程度。结果:细胞总RNA分别从8个只有肺癌组织标本和12对同时获取的肺癌组织及非癌肺组织标本中成功提取,然后用变性凝胶电泳鉴定...

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Published in癌症 Vol. 20; no. 3; pp. 274 - 278
Main Authors 毛友生, Austin Doyle, Weidong Yang, Yuetong Wei, Mark J.Krasna, Douglas D.Ross
Format Journal Article
LanguageChinese
Published 中国医学科学院,中国协和医科大学肿瘤医院胸外科,%Greenebaum Cancer Center, University of Maryland Medical School, Baltimore, Maryland 21201, USA 2001
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Abstract R734.2; 目的:进一步研究从人类乳腺癌细胞株(MCF-7/AdrVp)中克隆出的、对蒽环类抗癌药耐药的BCRP基因在非癌肺组织及非小细胞肺癌组织中的表达。方法:从术后即刻获得的正常肺组织和有活力的非小细胞肺癌组织新鲜标本中及时提取细胞总RNA,通过RT-PCR及PCR法获得并扩增BCRP基因的cDNA产物,再经凝胶电泳分离BCRP基因cDNA带,然后再通过转膜技术把这些BCRP基因cDNA产物转移到杂交膜上,用Southernblot杂交技术检测BCRP基因的表达程度。结果:细胞总RNA分别从8个只有肺癌组织标本和12对同时获取的肺癌组织及非癌肺组织标本中成功提取,然后用变性凝胶电泳鉴定提取RNA的质量。其中4例标本因术中缺血时间太长,或有坏死炎症或术前放化疗等原因,所提RNA有降解而放弃进一步的检测。通过RT-PCR及PCR方法从16例可用标本中获得并扩增BCRP基因的cDNA产物,再通过转膜及Southernblot杂交技术,最终发现BCRP基因在正常肺组织中均有不同程度的表达(10/10),而在非小细胞肺癌组织标本中只有一半病人的标本有不同程度的表达(8/16)。结论:BCRP基因可能是一个在非癌肺组织中常规表达的基因,而在部分病人的肺癌组织中可能此基因有缺失或被抑制。因此,在部分非小细胞肺癌患者中如用蒽环类抗癌药(如阿霉素,柔红霉素等)化疗时,可能会诱导BCRP基因的过度表达而产生对此类药物的耐药现象。
AbstractList R734.2; 目的:进一步研究从人类乳腺癌细胞株(MCF-7/AdrVp)中克隆出的、对蒽环类抗癌药耐药的BCRP基因在非癌肺组织及非小细胞肺癌组织中的表达。方法:从术后即刻获得的正常肺组织和有活力的非小细胞肺癌组织新鲜标本中及时提取细胞总RNA,通过RT-PCR及PCR法获得并扩增BCRP基因的cDNA产物,再经凝胶电泳分离BCRP基因cDNA带,然后再通过转膜技术把这些BCRP基因cDNA产物转移到杂交膜上,用Southernblot杂交技术检测BCRP基因的表达程度。结果:细胞总RNA分别从8个只有肺癌组织标本和12对同时获取的肺癌组织及非癌肺组织标本中成功提取,然后用变性凝胶电泳鉴定提取RNA的质量。其中4例标本因术中缺血时间太长,或有坏死炎症或术前放化疗等原因,所提RNA有降解而放弃进一步的检测。通过RT-PCR及PCR方法从16例可用标本中获得并扩增BCRP基因的cDNA产物,再通过转膜及Southernblot杂交技术,最终发现BCRP基因在正常肺组织中均有不同程度的表达(10/10),而在非小细胞肺癌组织标本中只有一半病人的标本有不同程度的表达(8/16)。结论:BCRP基因可能是一个在非癌肺组织中常规表达的基因,而在部分病人的肺癌组织中可能此基因有缺失或被抑制。因此,在部分非小细胞肺癌患者中如用蒽环类抗癌药(如阿霉素,柔红霉素等)化疗时,可能会诱导BCRP基因的过度表达而产生对此类药物的耐药现象。
Abstract_FL Objective:This study was designed to investigate BCRP gene expression in normal lung and in non-small cell lung cancer tissue. Methods:RNA was extracted immediately from the fresh normal lung tissue and viable tumor tissue harvested from surgically resected specimens of non-small cell lung cancer patients. cDNA of BCRP gene was prepared by RT-PCR and then amplified by PCR. The cDNA products from those specimens were transferred to blotting membrane through electrophoresis and transferring technique. Southern blot hybridization was eventually performed to detect the expression of BCRP gene. Results:RNAs were extracted from 8 tumor tissues and 12 pairs of tumor tissue and normal lung tissue were harvested from the same lung. Four patients’RNA samples with poor quality due to degrading were discarded. cDNA products of BCRP gene were obtained by RT-PCR and amplified by PCR in the remain 16 patients’RNA samples. Through Southern blot hybridization, BCRP gene was found to have slight expression in various amounts in all normal lung tissue(10/10) and only in half of tumor tissue samples (8/16). Conclusion:BCRP gene is slightly expressed in a variable amount in all normal lung tissue and only in half of tumor tissue for this group of patients with non-small cell lung cancer. Therefore chemotherapy with anthracycline may possibly induce the overexpression of BCRP gene and at last to develop multidrug resistance.
Author 毛友生
Austin Doyle
Weidong Yang
Mark J.Krasna
Yuetong Wei
Douglas D.Ross
AuthorAffiliation 中国医学科学院,中国协和医科大学肿瘤医院胸外科,%Greenebaum Cancer Center, University of Maryland Medical School, Baltimore, Maryland 21201, USA
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Austin Doyle
Weidong Yang
Mark J.Krasna
Yuetong Wei
Douglas D.Ross
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Keywords BCRP基因
多药耐药
化疗
肺肿瘤
非小细胞肺癌
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