Liao Hehe, Zhao Xixi, Qu Jinkun, Zhang Jia, Cai Hui
Second Department of Thoracic Surgery, The First Affiliated Hospital of Xi'an Jiaotong University 277 West Yanta Road, Xi'an 710061, Shaanxi, China.
Department of Oncology, The Second Affiliated Hospital of Xi'an Jiaotong University 157 West Fifth Street, Xi'an, 710004, Shaanxi, China.
Int J Clin Exp Med. 2015 Jul 15;8(7):10714-22. eCollection 2015.
Matrine has been proved to inhibit proliferation and induce apoptosis of human lung cancer cells. However, less studies involved in evaluating the effects and mechanism of matrine in cell migration and invasion of lung cancer. This study was aim to investigate the involvement of miR-133a in matrine's anti-invasion and anti-metastasis in lung cancer. MTT assay was used to assess the inhibition of proliferation effects of matrine in NCI-H1299 cells. Migration and invasion abilities of NCI-H1299 cells were investigated by Transwell assays. Expression of miR-133a was detected by real-time PCR. Anti-miR technique was applied to inhibit miR-133a in matrine treated HCI-H1299 cells. Real-time PCR and Western blotting were performed to evaluate the activation of EGFR/Akt/MMP-9 pathway. As results, matrine treatment significantly inhibited proliferation, migration and invasion of NCI-H1299 cells in a concentration-dependent manner, accompanied by significantly elevation of miR-133a expression. However, matrine failed to inhibit the metastatic ability when cells transfected with anti-miR-133a. Matrine treatment also suppressed activation of EGFR/Akt/MMP-9 pathway. The inhibitory effects of matrine on activation of EGFR pathway were also reversed by anti-miR-133a transfection in NCI-H1299 cells. In conclusion, matrine inhibited the invasion and metastasis of lung cancer cell by elevating expression of miR-133a which further suppressed activation of EGFR/Akt/MMP-9 pathway.
苦参碱已被证明可抑制人肺癌细胞的增殖并诱导其凋亡。然而,关于评估苦参碱对肺癌细胞迁移和侵袭影响及机制的研究较少。本研究旨在探讨miR-133a在苦参碱抗肺癌侵袭和转移中的作用。采用MTT法评估苦参碱对NCI-H1299细胞增殖的抑制作用。通过Transwell实验研究NCI-H1299细胞的迁移和侵袭能力。采用实时定量PCR检测miR-133a的表达。应用抗miR技术抑制苦参碱处理的HCI-H1299细胞中的miR-133a。通过实时定量PCR和蛋白质免疫印迹法评估EGFR/Akt/MMP-9信号通路的激活情况。结果显示,苦参碱处理以浓度依赖的方式显著抑制NCI-H1299细胞的增殖、迁移和侵袭,同时miR-133a表达显著升高。然而,当细胞转染抗miR-133a后,苦参碱未能抑制其转移能力。苦参碱处理还抑制了EGFR/Akt/MMP-9信号通路的激活。在NCI-H1299细胞中,抗miR-133a转染也逆转了苦参碱对EGFR信号通路激活的抑制作用。综上所述,苦参碱通过提高miR-133a的表达抑制肺癌细胞的侵袭和转移,进而抑制EGFR/Akt/MMP-9信号通路的激活。