【摘要】 目的:探讨土木香内酯(alantolactone,ALT)对人骨肉瘤143B细胞增殖、迁移、侵袭和凋亡的影响及其作用机制。方法:用不同浓度(0、4、6、8、10μmol/L)的ALT处理人骨肉瘤143B细胞后,用结晶紫染色法和MTT实验检测细胞的增殖能力,用划痕愈合实验、Transwell小室法、Hoechst33258染色法分别检测细胞的迁移、侵袭和凋亡水平,用qPCR及Western blotting(WB)分别检测细胞中E-cadherin和N-cadherin、caspase-3、cleaved-caspase-3(c-caspase-3)、PARP和cleaved-PARP(c-PARP)mRNA和蛋白表达水平。用荧光素酶报告基因实验检测T细胞淋巴因子或淋巴增强因子(T cell lymphocyte factor/lymphoid enhancer factor,TCF/LEF)转录活性,qPCR及WB检测β-catenin及MMP-7、c-Myc mRNA和蛋白表达水平。结果:ALT能够抑制骨肉瘤143B细胞的增殖、迁移和侵袭,同时促进细胞凋亡(P<0.05或P<0.01)。经8、10μmol/L ALT处理后,143B细胞中PARP m RNA和蛋白水平显著上调,c-caspase-3和c-PARP蛋白水平表达增加(均P<0.05);E-cadherin mRNA和蛋白水平表达上调、N-cadherin mRNA和蛋白表达水平下调,同时TCF/LEF转录活性明显下降(P<0.05或P<0.01),β-catenin、MMP-7和c-Myc mRNA和蛋白表达水平显著下调(P<0.05或P<0.01)。结论:ALT通过抑制Wnt/β-catenin信号通路的活性从而抑制人骨肉瘤143B细胞增殖、迁移和侵袭,并促进其凋亡。更多还原

【Abstract】 Objective: To investigate the effect of alantolactone(ALT) on proliferation, migration, invasion and apoptosis of human osteosarcoma 143 B cells and the underlying mechanism. Methods: Osteosarcoma 143 B cells were treated with different concentrations of ALT(0, 4, 6, 8, 10 μmol/L). Then, the cell proliferation ability was detected by crystal violet staining and MTT assay, cell migration was determined by Wound-healing test, cell invasion was analyzed by Transwell assay and cell apoptosis rate was detected by Hoechst33258 staining. The mRNA and protein expressions of E-cadherin, N-cadherin, caspase-3, cleaved caspase-3(c-caspase-3),poly ADP-ribose polymerase(PARP) and cleaved PARP(c-PARP) in 143 B cells were detected by qPCR and Western blotting(WB),respectively. TCF/LEF(T cell lymphocyte factor/lymphoid enhancer factor) transcriptional activity was examined with Luciferase reporter gene assay. The mRNA and protein expressions of β-catenin as well as MMP-7 and c-Myc were detected by qPCR and WB,respectively. Results: ALT inhibited proliferation, migration and invasion of osteosarcoma 143 B cells and promoted apoptosis(P<0.05 or P<0.01). After the treatment with ALT at 8, 10 μmol/L, the mRNA and protein expressions of E-cadherin and PARP, as well as the protein expressions of c-caspase-3 and c-PARP were up-regulated, while the mRNA and protein expressions of N-cadherin were downregulated(P<0.05 or P<0.01); At the same time, the TCF/LEF transcriptional activity and the mRNA and protein expressions of β-catenin,MMP-7 and c-Myc were significantly down-regulated(P<0.05 or P<0.01). Conclusion: ALT may inhibit the proliferation, migration and invasion and promote cell apoptosis possibly through suppressing Wnt/β-catenin signaling pathway in osteosarcoma 143 B cells.更多还原