Department of Science in Korean Medicine, Kyung Hee University, 24 Kyungheedae-ro, Dongdaemun-gu, Seoul 02447, Republic of Korea.
College of Korean Medicine, Dongguk University, 32 Dongguk-ro, Ilsandong-gu, Goyang-si, Gyeonggi-do 10326, Republic of Korea.
Life Sci. 2021 Nov 1;284:119893. doi: 10.1016/j.lfs.2021.119893. Epub 2021 Aug 26.
Tumor cells metastasis as well as proliferation are important factors that can substantially determines the prognosis of cancer. In particular, epithelial-mesenchymal transition (EMT) is key phenomena which can cause tumor cell transition into other organs by promoting the disruption of the cell-cell junctions. Because oxymatrine (OMT) have been reported to attenuate the tumor growth, we investigated whether OMT can down-regulate EMT process in tumor cells. We also focused on transforming growth factor-β (TGF-β)-induced EMT process because EMT process can be significantly induced by this growth factor.
The cell viability was measured by MTT and real time cell analysis (RTCA) assay. The expression levels of various proteins involved in the regulation of EMT and Akt/mTOR/PI3K signaling pathway were evaluated by Western blot analysis. mRNA levels of several important EMT markers were analyzed by reverse transcription polymerase chain reaction (RT-PCR). The effects of OMT on the cellular invasion and migration were evaluated by RTCA, wound healing assay, and boyden chamber assays.
OMT suppressed the expression of both constitutive and TGF-β-induced mesenchymal markers, such as fibronectin, vimentin, MMP-9, MMP-2, N-cadherin, Twist, and Snail, but induced the levels of epithelial markers. Moreover, OMT down-regulated oncogenic PI3K/Akt/mTOR pathways which lead to a significant attenuation of invasive and migratory potential of lung cancer cells.
Overall, our study established a novel anti-metastatic role of OMT against human lung cancer cells.
肿瘤细胞的转移和增殖是决定癌症预后的重要因素。特别是上皮间质转化（EMT）是一种关键现象，它可以通过促进细胞间连接的破坏，使肿瘤细胞向其他器官转移。由于氧化苦参碱（OMT）已被报道能抑制肿瘤生长，我们研究了 OMT 是否能下调肿瘤细胞中的 EMT 过程。我们还专注于转化生长因子-β（TGF-β）诱导的 EMT 过程，因为这种生长因子可以显著诱导 EMT 过程。
通过 MTT 和实时细胞分析（RTCA）测定细胞活力。通过 Western blot 分析评估参与 EMT 调节和 Akt/mTOR/PI3K 信号通路的各种蛋白质的表达水平。通过逆转录聚合酶链反应（RT-PCR）分析几个重要 EMT 标志物的 mRNA 水平。通过 RTCA、划痕愈合试验和 Boyden 室试验评估 OMT 对细胞侵袭和迁移的影响。
OMT 抑制了组成型和 TGF-β诱导的间充质标志物的表达，如纤连蛋白、波形蛋白、MMP-9、MMP-2、N-钙粘蛋白、Twist 和 Snail，但诱导了上皮标志物的水平。此外，OMT 下调致癌性的 PI3K/Akt/mTOR 通路，从而显著抑制肺癌细胞的侵袭和迁移潜力。
总的来说，我们的研究确立了 OMT 抑制人类肺癌细胞转移的新的抗肿瘤作用。
