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目的:将多药耐药基因(mdr1)转入CIK细胞,观察转染前后其对紫杉醇的耐药性及对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细胞对紫杉醇敏感性的变化,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蛋白的表达的变化;通过计算瘤重抑制率(TWI)检测转染前后的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,同时P-gp的表达较转染前及转染空质粒的CIK细胞均有显著增高(P<0.05),转染后的CIK细胞对紫杉醇的耐药性有显著提高,但对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 transfect multidrug resistance gene (mdr1) into CIK cells and observe the effects of mdr1 on paclitaxel resistance and the cytotoxicity of Lewis lung carcinoma cells before and after transfection. Methods: CIK cells were cultured in vitro and CIK cells were transfected with the recombinant plasmids of mdr1 during logarithmic growth phase. The expression of drug resistance genes was detected by RT-PCR. The sensitivity of CIK cells to paclitaxel was detected by MTT assay. The cytotoxic activity of CIK cells on Lewis lung cancer cells before and after staining was detected by Western blot. The expression of P-gp protein encoded by mdr1 was detected by Western blot. The effect of CIK cells before and after transfection on Lewis lung carcinoma Tumor inhibition. Results: The expression of mdr1 mRNA in CIK cells transfected with mdr1 was significantly higher than that in CIK cells transfected with empty plasmid (P <0.05), and the expression of P-gp was significantly increased in CIK cells transfected with mdr1 There was no significant change in the killing activity of Lewis lung cancer cells (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.