论文部分内容阅读
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，在部分非小细胞肺癌患者中如用蒽环类抗
Objective: To further study the expression of anthracycline-resistant BCRP gene cloned from human breast cancer cell line (MCF-7/AdrVp) in non-cancer lung tissue and non-small cell lung cancer tissue. METHODS: Total RNA was extracted from fresh specimens of normal lung tissue and viable non-small cell lung cancer tissue immediately after surgery. The cDNA product of BCRP gene was obtained and amplified by RT-PCR and PCR, and then was subjected to gel. The cDNA bands of BCRP gene were separated by electrophoresis, and then these BCRP gene cDNA products were transferred to the hybrid membrane by transfer membrane technology. The expression of BCRP gene was detected by Southern blot hybridization. RESULTS: Total cellular RNA was successfully extracted from 8 lung cancer-only specimens and 12 pairs of simultaneously acquired lung cancer tissues and non-cancer lung tissue specimens, and then the quality of the extracted RNA was identified by denaturing gel electrophoresis. Four of the specimens were deprived of further tests because of the long ischemia time during the operation, necrosis or preoperative radiotherapy and chemotherapy. The cDNA product of BCRP gene was obtained and amplified from 16 available specimens by RT-PCR and PCR. Finally, the BCRP gene was found to have different degrees of expression in normal lung tissues by transfer membrane and Southern blot hybridization technology (10) Only 10% of patients in non-small cell lung cancer tissues had different degrees of expression (8/16). CONCLUSIONS: The BCRP gene may be a gene that is normally expressed in non-cancerous lung tissues, but it may be absent or suppressed in some patients’ lung cancer tissues. Therefore, in some patients with non-small cell lung cancer, such as using anthracycline