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

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 Southern blot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 Southern blot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。

EXPRESSION OF BCRP GENE IN THE NORMAL LUNG TISSUE AND LUNG CANCER

Objective: To investigate the expression of novel multidrug resistance transporter (BCRP gene) from human MCF-7/AdrVp breast cancer cells in normal lung tissue and non-small lung cancer tissue. Methods: RNA was extracted immediately from 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 was then amplified by PCR. cDNA products from those specimens were transferred to blotting membrane through electrophoresis and transferring technique and southern blot hybridization was eventually performed to detect the expression of BCRP gene. Results: RNA were extracted from 8 tumor tissue alone and 12 pairs of tumor tissue and normal lung tissue 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 were then amplified by PCR in the remain 16 patients’ RNA samples. Through southern blot hybridization, BCRP gene was found to be slightly expressed in various amounts in all normal lung tissue (10/10) and only in a half of tumor tissue samples (8/16). Conclusion: BCRP gene is slightly expressed in different amount in all normal lung tissue and only in a half of tumor tissue of non-small cell lung cancer patients. It is possible to induce it’s overexpression and to develop multidrug resistance during chemotherapy if using anthracycline anticancer drugs.