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目的观察化疗增敏剂对多药耐药性(Multi-drug resistance,MDR)肿瘤细胞P-糖蛋白(P-glycoprotein,P-gp)的影响及伴随99Tcm-MI BI摄取动力学的变化,以建立用99Tcm-MIBI来评价化疗增敏剂效果的方法。方法MDR人乳腺癌细胞MCF-7/Adr 37℃培养。(1)实验组和对照组细胞培养基中分别加入化疗增敏剂维拉帕米(10μmol/L) 和等体积培养液DMEM。99Tcm-MIBI与细胞共同孵育2 h后收集细胞,测定放射性活度和P-gp表达水平。(2)维拉帕米(10μmol/ L)加入细胞培养基中,与细胞孵育若干时间后99Tcm-MIBI加入细胞培养基中,与细胞一起培养2 h后收集细胞。测定放射性活度和P-gp表达水平。结果(1)维拉帕米作用2h后,细胞摄取99Tcm-MIBI较对照组显著增加(t=2.33,P<0.05),但P-gp表达水平差异无显著性(P> 0.05)。(2)肿瘤细胞摄取99Tcm-MIBI随维拉帕米作用时间的延长而增加,各时间点UR差异有显著性(F=58.2,P<0.05),并且, VRP作用时间在80 min内时,肿瘤细胞摄取99Tcm-MIBI与P-gp的表达水平无相关性(r=0.16,P>0.05),VRP作用时间大于8 h 时,肿瘤细胞摄取99Tcm-MIBI与P-gp的表达水平呈负相关(r=-0.73,P<0.01)。结论化疗增敏剂能够影响P-gp过度表达所致MDR肿瘤细胞对99Tcm-MIBI的摄取。
Objective To observe the effects of chemosensitizers on P-glycoprotein (P-gp) and the uptake kinetics of 99Tcm-MI BI in multi-drug resistance (MDR) The method of evaluating the effect of chemosensitizer by 99Tcm-MIBI was established. Methods MDR human breast cancer cells MCF-7 / Adr cultured at 37 ℃. (1) Verapamil (10μmol / L) and equal volume of culture medium DMEM were added into the cell culture medium of experimental group and control group respectively. After incubation with 99Tcm-MIBI for 2 h, the cells were collected and the radioactivity and the expression of P-gp were measured. (2) Verapamil (10μmol / L) was added to the cell culture medium, 99Tcm-MIBI was added to the cell culture medium after incubation with the cells for several hours, and the cells were collected after cultured with the cells for 2 hours. Radioactivity and P-gp expression levels were determined. Results (1) The cell uptake of 99Tcm-MIBI in verapamil group increased significantly (t = 2.33, P <0.05) after 2 hours of treatment with verapamil, but there was no significant difference in the expression level of P-gp between two groups (P> 0.05). (2) The uptake of 99Tcm-MIBI by tumor cells increased with the prolongation of the effect of verapamil, and the difference of UR was significant at each time point (F = 58.2, P <0.05). When the action time of VRP was within 80 min, The uptake of 99Tcm-MIBI by tumor cells had no correlation with the expression of P-gp (r = 0.16, P> 0.05). When VRP was longer than 8 h, the expression of 99Tcm-MIBI was negatively correlated with the expression of P-gp (r = -0.73, P <0.01). Conclusion Chemosensitizers can affect the uptake of 99Tcm-MIBI in MDR tumor cells induced by P-gp overexpression.