摘要
目的探究端锚聚合酶抑制剂XAV-939对人骨肉瘤细胞增殖迁移的影响及作用机制。方法体外培养人骨肉瘤MG-63细胞,根据XAV-939终浓度的不同分为空白对照组、实验1组(5μmol/L)、实验2组(10μmol/L)、实验3组(15μmol/L)。采用CCK-8增殖实验测定对照组与实验组MG-63细胞增殖情况,并计算抑制率;采用伤口愈合试验和Transwell侵袭实验检测对照组与实验组MG-63细胞的迁移侵袭能力;采用蛋白免疫印记实验检测对照组与实验组MG-63细胞中β-catenin、基质金属蛋白酶Ⅱ(MMP-2)蛋白以及上皮间质转换标志物E-cadherin和N-cadherin蛋白的表达水平的变化。结果 CCK-8增殖实验结果显示,3组实验组细胞的增殖抑制率均明显高于空白对照组(P <0.01),且3组实验组中随着XAV-939终浓度的提高,细胞增殖抑制率逐渐升高,差异有统计学意义(P <0.05);培养48 h时,实验组细胞增殖抑制率均高于培养24 h时(P <0.05),而对照组细胞培养24 h与48 h时相比,细胞增殖抑制率差异无统计学意义(P> 0.05)。伤口愈合试验和Transwell侵袭实验结果显示,实验组细胞迁移、侵袭能力明显低于空白组(P <0.05),3组实验组中随着XAV-939终浓度的提高,细胞的迁移、侵袭能力逐渐降低,差异有统计学意义(P <0.05)。蛋白免疫印记实验结果显示,与对照组相比,β-catenin、MMP-2、N-cadherin蛋白在实验组中表达水平下降,而E-cadherin蛋白表达水平上升,差异有统计学意义(P <0.05)。结论 XAV-939能有效抑制人骨肉瘤MG-63细胞增殖侵袭能力,其作用机制可能与抑制Wnt/β-catenin信号通路进而逆转上皮间质转换过程有关,XAV-939有望成为治疗人骨肉瘤的靶向药物。
Objective To investigate into the effect of tankyrase inhibitor XAV-939 on the proliferation and migration of human osteosarcoma cells and its mechanism.Methods Human osteosarcoma MG-63 cells were cultured in vitro and divided into a blank control group,experimental group 1(5μmol/L),experimental group2(10μmol/L),and experimental group 3(15μmol/L).The proliferation of MG-63 cells in the control group and experimental group was measured by CCK-8 proliferation assay and the inhibition rate was calculated.Wound healing assay and Transwell invasion assay were used to detect the migration and invasion ability of MG-63 cells in the control and experimental groups.Protein immunization,the expression ofβ-catenin,MMP-2 protein and epithelial-mesenchymal transition markers E-cadherin and N-cadherin in MG-63 cells in the control group and experimental group were detected by Western blotting.Results The results of the proliferation assay of CCK-8 showed that the proliferation inhibition rates of the three experimental groups were significantly higher than that of the blank control group(P<0.01).With the increase of the final concentration of XAV-939 in the three experimental groups,the cell proliferation was inhibited(P<0.05).The inhibitory rate of cell proliferation in the experimental group at 48 h was significantly higher than that in at 24 h(P<0.05),the inhibition rate of cell proliferation in control group at 48 h was not significantly different from that at 24 h(P>0.05).Wound healing test and Transwell invasion assay showed that the cell migration and invasion ability of the experimental group were significantly lower than that of the blank group(P<0.05).With the increase of the final concentration of XAV-939 in three experimental groups,the migration and invasion ability of cells gradually decreased,the difference being statistically significant(P<0.05).Western blotting results showed that,compared with the control group,the expression ofβ-catenin,MMP-2 and N-cadherin in the experimental group decreased,while the
作者
赵珂
韩兴文
吴永娜
李二亮
王文己
Zhao Ke;Han Xing-wen;Wu Yong-na;Li Er-liang;Wang Wen-ji(Department of Orthopaedics,Gansu Provincial Hospital,Lanzhou730000,China;Department of Orthopaedics,The First Hospital of Lanzhou University,Lanzhou730000,China;Gansu Provincial Key Laboratory of Biotherapy and Regenerative Medicine,Lanzhou730000,China)
出处
《兰州大学学报（医学版）》
CAS
2019年第2期61-66,共6页
Journal of Lanzhou University（Medical Sciences）
基金
甘肃省自然科学基金项目(160RJZA126)
甘肃省科技计划资助项目(17JR5RA196)
