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目的 化疗是脑胶质瘤术后综合治疗的主要措施,由于胶质瘤细胞存在原发或继发的化疗药物耐受性,致使化疗失败。本文试图寻求一种简便、敏感、准确的检测方法.确定肿瘤细胞的耐药程度,为合理制定化疗方案提供依据。方法 采用软琼脂糖克隆法检测14例脑胶质瘤和u251MG细胞株对长春新碱的药物敏感性,通过VCRIC50估价肿瘤的耐药程度。同时采用以cRNA为内参照的RT-PCR方法同步检测胶质瘤MDR1基因mRNA含量,根据单位总RNA含MDR1,mRNA的分子数确定耐药程度。结果 以U251MG细胞株为对照,确定脑胶质瘤的耐药程度,VCRlC50在1.73～28.80ng之间。MDR1 mRNA分数在0.80～14.59×1O~5之间。两种方法相关性比较,r=0.77。琼脂糖克隆法其中度以上耐药的检出率为50％,而应用RT-PCR方法的检出率为33.50％。系统化疗后肿瘤细胞的MDR1,mRNA含量明显增高。结论 琼脂克隆法可直接准确地检测脑胶质瘤的化疗耐药性.有较高的临床应用价值。
The purpose of chemotherapy is the main measures of comprehensive treatment of glioma postoperatively, due to the presence of primary or secondary chemotherapy drug resistance of glioma cells, resulting in the failure of chemotherapy. This article attempts to seek a simple, sensitive and accurate detection methods to determine the degree of resistance of tumor cells, to provide a basis for the rational formulation of chemotherapy. Methods The sensitivity of vincristine to 14 gliomas and u251MG cells was detected by soft agarose gel electrophoresis. The drug resistance of vincristine was assessed by VCRIC50. At the same time, the mRNA level of MDR1 gene in glioma was detected synchronously by RT-PCR with cRNA as an internal reference, and the drug resistance was determined according to the total number of molecules containing MDR1 and mRNA in total RNA. Results U251MG cell line as a control to determine the degree of glioma resistance, VCRlC50 between 1.73 ~ 28.80ng. The MDR1 mRNA scores ranged from 0.80 to 14.59 × 10 5. Correlation between the two methods, r = 0.77. Agarose cloning method was moderately resistant to more than 50% of the detection rate, and the application of RT-PCR method was detected in 33.50%. After chemotherapy, tumor cells MDR1, mRNA content was significantly increased. Conclusion agar cloning method can directly and accurately detect the chemoresistance of glioma, and has high clinical value.