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目的:将多药耐药基因(mdr1)转入CIK细胞,观察转染前后其对顺铂的耐药性及对Lewis肺癌细胞杀伤活性的影响。方法:常规方法培养CIK细胞,在细胞对数生长期,将mdr1的重组质粒转染CIK,通过RT-PCR鉴定耐药基因表达;MTT法检测CIK细胞对顺铂敏感性的变化,同时检测转染前后CIK细胞对Lewis肺癌细胞的杀伤活性变化;Western blot检测细胞中mdr1编码的P-gp蛋白表达的变化;通过计算瘤重抑制率(TWI)检测转染前后CIK细胞对Lewis肺癌移植瘤的抑制作用。结果:转染mdr1后的CIK细胞mdr1 mRNA阳性,在转染后的CIK细胞中,P-gp的表达较转染前CIK细胞及转染空质粒的CIK细胞明显增高,差异有统计学意义(P<0.05),转染mdr1基因后的CIK细胞对顺铂的耐药性明显提高,转染前后CIK细胞对Lewis肺癌细胞的杀伤活性无明显变化(P>0.05)。结论:将mdr1基因转入CIK细胞后,细胞获得了多药耐药性,同时保持了原有的对肿瘤细胞的杀伤活性。
OBJECTIVE: To transfer the multidrug resistance gene (mdr1) into CIK cells and observe its drug resistance to cisplatin and its effect on Lewis lung cancer cell killing activity before and after transfection. Methods: CIK cells were cultured by conventional methods. The mdr1 recombinant plasmids were transfected into CIK cells in the logarithmic growth phase, and the expression of drug-resistant genes was identified by RT-PCR. The sensitivity of CIK cells to cisplatin was detected by MTT assay. The cytotoxicity of CIK cells to Lewis lung cancer cells before and after dyeing was detected by Western blot. The expression of mdr1-encoded P-gp protein was detected by Western blot. The inhibitory rates of tumor weight (TWI) Inhibition. Results: The mdr1 mRNA of CIK cells transfected with mdr1 was positive. The expression of P-gp in transfected CIK cells was significantly higher than that of CIK cells and CIK cells transfected with empty plasmid before transfection P <0.05). CIK cells transfected with mdr1 gene showed a significant increase in the resistance to cisplatin. The cytotoxicity of CIK cells to Lewis lung cancer cells did not change significantly before and after transfection (P> 0.05). Conclusion: The mdr1 gene was transfected into CIK cells, the cells obtained multidrug resistance, while maintaining the original killing activity of tumor cells.