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目的探讨辐射促转染的多药耐药基因(mdr1)反义寡核苷酸(ASON)逆转肿瘤细胞KBv200的耐药效果。方法应用MTT法比较脂质体介导的ASON直接转染与联合辐射转染的情况下KBv200细胞对长春新碱(VCR)的半数抑制量(IC50),再利用RTPCR方法和流式细胞仪检测两种不同的转染方法对KBv200细胞的mdr1mRNA及其表达产物P糖蛋白(Pgp)的调控情况。采用耐药细胞株KBv200移植于裸鼠皮下,肿瘤局部2Gy60Coγ射线照射后1h,瘤内注射脂质体介导的mdr1ASON,经VCR联合化疗。结果联合照射的ASON组明显优于未照射组(P<0.05)。KBv200细胞的IC50、mdr1mRNA的表达水平及Pgp的阳性率,均明显低于未照射组(P<0.05)。联合照射组与未照射组肿瘤体积和肿瘤重量差异有统计学意义(P<0.01)。结论辐射促转染的mdr1ASON对肿瘤细胞具有较强的耐药逆转作用。
Objective To investigate the multidrug resistance (mdr1) antisense oligonucleotide (ASON) induced by radiation to reverse the drug resistance of tumor cells KBv200. Methods MTT assay was used to compare the IC50 of vincristine (VCR) in KBv200 cells transfected with liposome-mediated ASON direct transfection combined with radiation. Then RTPCR and flow cytometry Regulation of mdr1 mRNA and P glycoprotein (Pgp) expression in KBv200 cells by two different transfection methods. The drug-resistant cell line KBv200 was transplanted subcutaneously in nude mice. The tumors were injected with liposome-mediated mdr1ASON intratumorally 1 h after 2Gy60Coγ-ray irradiation and were treated with VCR combined with chemotherapy. Results The combined ASON group was significantly better than the non-irradiated group (P <0.05). The expression of IC50, mdr1 mRNA and the positive rate of Pgp in KBv200 cells were significantly lower than those in non-irradiated group (P <0.05). There were significant differences in tumor volume and tumor weight between the combined irradiation group and the non-irradiated group (P <0.01). Conclusion The radiation-transfected mdr1ASON has a strong reversal effect on tumor cells.