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目的:探讨印记基因SLC22A18(solute carrier family 22,member 18)对人胶质瘤U251细胞化疗药物敏感性的影响及耐药机制的研究。方法:采用脂质体转染法将携带有SLC22A18基因的重组质粒pIRES2-EGFP-SLC22A18转入U251细胞;分别采用RT-PCR和蛋白质印迹法检测SLC22A18 mRNA及蛋白在转染后U251细胞中的表达情况;CCK-8法检测细胞对化疗药物敏感性的变化;FCM法检测SLC22A18表达对细胞凋亡以及对多柔比星在细胞内蓄积浓度的影响。结果:转染SLC22A18基因的U251细胞中有SLC22A18 mRNA及其蛋白的表达;SLC22A18基因转染组U251细胞与对照组细胞比较,U251细胞对紫杉醇的敏感性下降,半数抑制浓度(halfinhibitory concentration,IC50)值上升(t=3.23,P<0.05),对替莫唑胺的敏感性上升,IC50值下降(t=4.28,P<0.05)。SLC22A18基因转染组和空质粒转染组细胞凋亡率分别为(41.35±4.98)%和(6.25±0.82)%,差异有统计学意义(t=12.05,P<0.01)。SLC22A18表达使细胞内多柔比星蓄积浓度明显下降(t=4.25,P<0.05)。结论:SLC22A18表达使人胶质瘤U251细胞对紫杉醇的敏感性下降,对替莫唑胺的敏感性提高,并可能通过降低细胞内化疗药物蓄积产生耐药性。
Objective: To investigate the effect of SLC22A18 (member 18) on chemosensitivity of human glioma U251 cells and its mechanism of drug resistance. Methods: The recombinant plasmid pIRES2-EGFP-SLC22A18 carrying SLC22A18 gene was transfected into U251 cells by lipofection method. The expression of SLC22A18 mRNA and protein in transfected U251 cells was detected by RT-PCR and Western blot respectively The changes of cell sensitivity to chemotherapeutic drugs were detected by CCK-8 assay. The effect of SLC22A18 expression on cell apoptosis and the concentration of doxorubicin in cells were detected by FCM. Results: SLC22A18 mRNA and protein expression were found in U251 cells transfected with SLC22A18 gene. U251 cells transfected with SLC22A18 gene showed a decreased sensitivity to paclitaxel and a lower half-inhibitory concentration (IC50) (T = 3.23, P <0.05). The sensitivity to temozolomide increased and the IC50 decreased (t = 4.28, P <0.05). The apoptosis rates of SLC22A18 gene transfected group and empty plasmid transfected group were (41.35 ± 4.98)% and (6.25 ± 0.82)%, respectively, with statistical significance (t = 12.05, P <0.01). SLC22A18 expression significantly decreased the concentration of doxorubicin in the cells (t = 4.25, P <0.05). CONCLUSIONS: SLC22A18 expression decreases the sensitivity of human glioma U251 cells to paclitaxel, increases the sensitivity to temozolomide, and may result in drug resistance by reducing the accumulation of intracellular chemotherapy drugs.