【摘要】 目的:探讨石蒜碱(lycorine)对人结肠腺癌LoVo细胞活力、凋亡和迁移侵袭能力的影响及其作用机制。方法:不同浓度(1～8μmol/L) lycorine处理LoVo细胞,CCK-8法检测细胞活力;划痕愈合和Transwell小室实验检测迁移和侵袭能力;Hoechst染色及流式细胞术实验检测凋亡改变;萤光素酶报告基因系统检测MAPK/ERK信号通路活化情况;同时采用Western blot法检测增殖细胞核抗原(proliferating cell nuclear antigen, PCNA)、上皮-间充质转化(epithelial-mesenchymal transition, EMT)相关标志物、凋亡相关蛋白及磷酸化ERK1/2蛋白水平的变化。结果:与空白对照组相比,lycorine处理组LoVo细胞的活力和迁移侵袭能力受到显著抑制(P<0.05),同时EMT相关标志物基质金属蛋白酶7(matrix metalloproteinase-7, MMP-7)和N-cadherin表达下调,E-cadherin表达上调(P<0.05),而凋亡细胞数及cleaved caspase-3无显著差异(P>0.05)。此外,lycorine处理组Elk1/SRF转录活性降低,ERK1/2磷酸化水平降低(P<0.05)。结论:石蒜碱可通过调控MAPK/ERK信号通路及抑制EMT过程LoVo细胞的活力及迁移侵袭能力,从而在体外主要发挥生长抑制的抗肿瘤作用。更多还原

【Abstract】 AIM: To investigate the effect of lycorine on the viability, migration, invasion and apoptosis of human colon adenocarcinoma LoVo cells. METHODS: After treatment with lycorine at different concentrations(1～8 μmol/L), the cell viability was measured by CCK-8 assay. Wound-healing and Transwell chamber assay were used to evaluate cell migration and invasion abilities. Hoechst staining and flow cytometry were adopted to test the changes of apoptosis. The activation of MAPK/ERK signaling pathway was determined by luciferase reporter system. The protein levels of proliferating cell nuclear antigen(PCNA), epithelial-mesenchymal transition(EMT)-related markers, apoptosis-related proteins, and p-ERK1/2 were detected by Western blot. RESULTS: Compared with blank control group, the viability, migration and invasion of LoVo cells treated with lycorine were significantly inhibited(P<0.05). The levels of EMT-related markers matrix metalloproteinase-7(MMP-7) and N-cadherin were down-regulated remarkably(P<0.05), and the change of E-cadherin was the opposite(P<0.05). However, the number of apoptotic cells and the protein level of cleaved caspase-3 had no obvious changes(P>0.05). Additionally, the transcriptional activity of Elk1/SRF the protein level of p-ERK1/2 were decreased(P<0.05). CONCLUSION: Lycorine inhibits the viability, migration and invasion of LoVo cells via regulating MAPK/ERK signaling pathway and inhibiting EMT process. In vitro, it mainly exerts anti-tumor activity through cytostatic effects.更多还原