论文部分内容阅读
目的 探讨免疫活性细胞对肿瘤细胞的杀伤活性与mdr1的关系。方法 利用特异性切割mdr1的ribozyme为工具 ,以表达mdr1的耐药细胞株KBv2 0 0 为靶细胞 ,采用脂质体转染技术 ,将含ribozyme的质粒pHβApr 1neo/ 5mR3及空载体pHβApr 1neo导入KBv2 0 0 及其亲本KB细胞内 ,运用North ernblotting、免疫组化方法观察ribozyme对mdr1mRNA及P 170的影响 ,采用MTT法检测了CD3AK、LAK对KB、KBv2 0 0 细胞株耐药逆转前后杀伤活性的变化。结果 含ribozyme的质粒pHβApr 1neo/ 5mR3及空载体pHβApr 1neo可以在KB、KBv2 0 0 细胞中稳定表达 ,ribozyme可以特异性地切割mdr1,导致KBv2 0 0 / 5mR3的mdr1mRNA含量下降 ,P 170表达减低 ,LAK和CD3AK对KBv2 0 0 的杀伤活性较对KB的强 ,MDR逆转后杀伤活性降至敏感株水平 ,LAK和CD3AK对KB/ 5mR3、KB/vec、KBv2 0 0 /vec的杀伤活性也较KB及KBv2 0 0 / 5mR3强。结论 mdr1 ribozyme在细胞内具有一定的逆转肿瘤多药抗性的生物学效应 ,LAK和CD3AK对MDR肿瘤细胞的杀伤活性与P糖蛋白表达相关
Objective To investigate the relationship between mdr1 and the killing activity of immunocompetent cells on tumor cells. Methods Using the ribozyme specifically cutting mdr1 as a tool, the resistant cell line KBv200 expressing mdr1 was used as a target cell. The plasmid pHβApr 1neo/ 5mR3 containing ribozyme and the empty vector pHβApr 1neo were introduced into KBv2 using liposome transfection technology. In 0 and its parental KB cells, Northern blotting and immunohistochemistry were used to observe the effects of ribozyme on mdr1 mRNA and P 170. MTT assay was used to detect the killing activity of CD3AK and LAK before and after drug resistance reversal in KB and KBv200 cell lines. Variety. Results The ribozyme-containing plasmid pHβApr 1neo/5mR3 and the empty vector pHβApr 1neo could be stably expressed in KB and KBv200 cells. ribozyme could specifically cleave mdr1, resulting in the decrease of mdr1 mRNA and the decrease of P 170 expression in KBv200/5mR3. The killing activity of LAK and CD3AK against KBv200 was stronger than that of KB. The reversal of MDR reduced the killing activity to sensitive strains. The killing activity of LAK and CD3AK against KB/5mR3, KB/vec, and KBv200/vec was also higher than that of KB. And KBv2 0 0 / 5mR3 strong. Conclusion The mdr1 ribozyme has a certain biological effect in reversing multidrug resistance of tumors. The killing activity of LAK and CD3AK on MDR tumor cells is related to the expression of P-glycoprotein.