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目的探讨塞来昔布增强口腔癌细胞化疗敏感性与其阻滞细胞周期进程及调节P-gp之间的相关性。方法塞来昔布10、20、40、80μmol/L和/或长春新碱0.375、0.75、1.5、3μmol/L处理KB/VCR细胞后,分别采用MTT法检测KB/VCR细胞生长抑制率;流式细胞术检测细胞周期分布情况;Western blot检测周期相关蛋白Cyclin D1、p21WAF1/CIP1及P-gp的蛋白表达。结果塞来昔布浓度较低时(<20μmol/L)对KB/VCR生长增殖无明显影响,但小剂量的塞来昔布10μmol/L可显著增强VCR对KB/VCR细胞的毒性作用,并呈时间依赖性。塞来昔布与长春新碱联合作用于KB/VCR细胞24、48、72 h后,其生长抑制率分别为(37.53±2.05)%、(46.67±3.17)%及(54.02±1.53)%,显著高于塞来昔布和长春新碱单独用药组(P均<0.01)。塞来昔布与VCR联合应用能改变细胞周期分布,与VCR组相比,联合用药组G0/G1期细胞数量增加(56.08±0.46)%,S期和G2/M期细胞数目降低[S期:(22.83±0.20)%;G2/M期:(21.09±0.66)%]。此外,与VCR组细胞相比,联合用药能显著降低Cyclin D1的表达,增强p21WAF1/CIP1的表达,并且Cyclin D1蛋白水平的降低及p21WAF1/CIP1蛋白水平的上升伴随着P-gp蛋白的下调。结论塞来昔布下调节P-gp而增强KB/VCR细胞对化疗药物的敏感性可能与Cyclin D1和p21WAF1/CIP1蛋白变化引起的细胞周期阻滞有关。
Objective To investigate the effect of celecoxib on the chemosensitivity of oral cancer cells to the cell cycle arrest and the regulation of P-gp. Methods KB / VCR cells were treated with celecoxib 10, 20, 40, 80μmol / L and / or vincristine 0.375,0.75,1.5 and 3μmol / L for KB / VCR. Western blot was used to detect the protein expression of Cyclin D1, p21WAF1 / CIP1 and P-gp. Results Low concentrations of celecoxib (<20 μmol / L) had no significant effect on the proliferation of KB / VCR cells. However, the low dose of celecoxib 10 μmol / L significantly increased the toxicity of VCR to KB / VCR cells Time-dependent. The inhibitory rates of celecoxib and vincristine on KB / VCR cells were (37.53 ± 2.05)%, (46.67 ± 3.17)% and (54.02 ± 1.53)%, respectively, Significantly higher than that of celecoxib and vincristine alone (all P <0.01). The combination of celecoxib and VCR could change the cell cycle distribution. Compared with the VCR group, the number of cells in the G0 / G1 phase increased (56.08 ± 0.46)% in the combination group and decreased in the S and G2 / M phases [S phase : (22.83 ± 0.20)%; G2 / M phase: (21.09 ± 0.66)%]. In addition, compared with VCR cells, the combination treatment can significantly reduce the expression of Cyclin D1 and enhance the expression of p21WAF1 / CIP1, and the decrease of Cyclin D1 protein and the increase of p21WAF1 / CIP1 protein accompanied with the down-regulation of P-gp protein. CONCLUSION: Celecoxib may increase the sensitivity of KB / VCR cells to chemotherapeutic drugs by down-regulating P-gp, which may be related to the cell cycle arrest induced by the changes of Cyclin D1 and p21WAF1 / CIP1 proteins.