多药耐药基因作用新发现.docVIP

多药耐药基因作用新发现 　　[摘要] 目的：观察被照射的小细胞肺癌细胞系在外照射前后其总RNA（核糖核酸）、MDR1 mRNA（多药耐药基因的信使核糖核酸）、药物敏感性和放射敏感性的变化，从而探讨多药耐药基因是否还有其他未被揭示的新作用。方法：采用GWGP80型远距离60Co治疗机对进入指数生长期的NCI-H446小细胞肺癌细胞系进行分次外照射。用Trizol分别提取未照射细胞（S-cell，Sc）和已完成全部外照射剂量的NCI-H446细胞（R-cell，Rc）的总RNA。采用RT-PCR（逆转录聚合酶链反应）法来确定两组细胞MDR1 mRNA表达的高低。通过加入丝裂霉素和逆转剂维拉帕米后的干扰来分别观察外照射前后NCI-H446细胞系存活率的变化。然后对Sc和Rc再给予不同剂量的外照射，1周后计算它们各自的集落形成率（PE）和存活率（S）。结果：在相同细胞数和相同体积的条件下，未照射组和照射组细胞总RNA的浓度分别为25.9 mg/L和16.6 mg/L；未照射组细胞其MDR1 cDNA（多药耐药基因的逆转录脱氧核糖核酸）/β-actin cDNA为1.078，照射组为1.338。在相同浓度的化疗药丝裂霉素的干扰下，照射组细胞的存活率均明显高于未照射组（P0.05）。在分别给予2 Gy和4 Gy的外照射后，照射组细胞的存活率（S）分别为（79.67±34.48）%和（23.73±8.62）%，而未照射组的分别为（51.24±11.88）%和（19.40±5.59）%，其差异有统计学意义（P 　　[关键词] 细胞系；小细胞肺癌；外照射；MDR1 mRNA ；药物敏感性；丝裂霉素；逆转作用；维拉帕米；放射敏感性 　　[中图分类号] R730.5 [文献标识码] A [文章编号] 1673-7210（2012）01（a）-005-05 　　 　　The new findings of multi-drug resistant (MDR) genes′s effect 　　SONG Peng, DUAN Yunyou, HUANG Youzhang, WU Haibo 　　Department of Quality Control, Navy General Hospital of PLA, Beijing 100048, China 　　[Abstract] Objective: To observe the change of the total RNA and MDR1 gene, the chemosensitivity and the radio-sensitivity of NCI-H 446 small cell lung cancer cell line, and to explore the other effect of multidrug resistant (MDR) genes. Methods: NCI-H 446 small cell lung cancer cell line which was exponential phase of growth was explored to 60Co treatment machine (GWGP80 type). The total RNA of the cells that were unirradiated (S-cell, Sc) and irradiated (R-cell, Rc) were isolated from homogenate samples by the acid guanidine thiocyanate-phenol chloroform method. The expression of MDR1 mRNA was assessed by qualitative RT-PCR assays. The survival rates of NCI-H446 cell line were respectively observed by the interference of different concentration of Mitomycin C and the reversal agent of verapamil before and after fractioned radiation treatment. The two kinds of cells were administered different doses of radiation treatment again. And then the plating efficiency (PE) and the survival rate (S)