血根碱对人肺腺癌细胞迁移、侵袭和Wnt/β-catenin信号通路的影响

杨佳; 陈旻; 李明花; 钦敬茹; 王中奇

doi:10.3971/j.issn.1000-8578.2019.18.1576

血根碱对人肺腺癌细胞迁移、侵袭和Wnt/β-catenin信号通路的影响

Effects of Sanguinarine on Invasion, Migration and Wnt/β-catenin Signaling Pathway of Lung Adenocarcinoma Cells

摘要

摘要:
目的探讨血根碱对人肺腺癌细胞迁移、侵袭的影响及其初步分子机制。
方法 MTT法检测血根碱对肺腺癌细胞活性的影响；利用划痕和Transwell小室实验分析血根碱对细胞迁移、侵袭的影响；RT-qPCR和免疫印迹法检测不同浓度血根碱对Wnt/β-catenin通路相关基因和核心蛋白的影响。
结果与空白对照组比，血根碱（1~10 μmol/L）呈浓度-时间依赖性地抑制人肺腺癌细胞的活力（P < 0.05）；低浓度1 μmol/L作用48 h后，A549和H1975细胞的迁移和侵袭能力明显减弱（P < 0.01）；血根碱（1、2.5、5 μmol/L）可显著下调Wnt/β-catenin通路中核心分子β-catenin及上游磷酸化GSK3β、DVL2蛋白表达，进而抑制下游靶基因TCF/LEF，CyclinD1的mRNA水平（P < 0.05），并呈一定的浓度依赖性。
结论血根碱可能通过下调Wnt/β-catenin信号通路起到抑制人肺腺癌细胞迁移、侵袭的作用。

Abstract:
Objective To investigate the effect of sanguinarine on the invasion and migration of lung adenocarcinoma cells and its preliminary molecular mechanism.
Methods MTT assay was used to observe the proliferative capacity of A549 and H1975 cells. Wound healing and Transwell assays were performed to determine the invasion and migration abilities of A549 and H1975 cells, respectively. RT-qPCR and Western blot were used to detect the mRNA and protein expression of core genes in Wnt/β-catenin signaling pathway.
Results Compared with the control group, sanguinarine(1-10μmol/L) could inhibit the activity of human lung adenocarcinoma cells in a concentration- and time- dependent manner (P < 0.05); the migration and invasion abilities of A549 and H1975 cells were significantly weakened by 1μmol/L sanguinarine after 48 h (P < 0.01); sanguinarine (1, 2.5, 5μmol/L) could significantly down-regulate the protein expression of β-catenin, phosphorylated GSK3β and DVL2 in Wnt/β-catenin pathway, thereby inhibiting the mRNA levels of TCF/LEF and CyclinD1, in a concentration-dependent manner (P < 0.05).
Conclusion Sanguinarine may inhibit the migration and invasion of human lung adenocarcinoma cells by down-regulating the Wnt/β-catenin signaling pathway.

HTML全文

参考文献(22)

施引文献

资源附件(0)