Abstract
Objective
Dihydroartemisinin (DHA), a predominant phytoconstituent in Artemisia annua L. (a plant widely used as a traditional medicine in China), inhibits lung tumorigenesis and metastasis. However, its anticancer effect against hepatocellular carcinoma has not yet been investigated. In this study, the anti-tumor potential of DHA was evaluated in vitro against the hepatocellular carcinoma HCCLM6 cell line.
Results
DHA (1–100 μM) treatment suppressed the cell proliferation in dose-dependently. In addition, expression of all genes, involved in cellular proliferation (e.g. E2F1, BCL2, PCNA, MKI67 and CCNE2) and cellular motility (e.g. DOCK1, ITGA2, VCL, MMP2, FN1), was significantly downregulated by DHA (50 and 100 μM). Global gene expression profile identified 1731 differentially expressed genes (DEGs); among them, 211 were up-regulated and 1520 were down-regulated. Besides, the extracellular matrix (ECM)-receptor interaction, focal adhesion, regulation of actin cytoskeleton and TNF pathways were enriched by DEGs. Based on the KEGG signal pathway enrichment, the FN1 and integrin-β1 could be a potential target for DHA for inhibiting proliferation. The expression of FN1 and integrin-β1 was further analyzed by the qPCR, immunohistochemistry and Western blot assay in vitro and in vivo. The results indicated that DHA decreased the FN1 and integrin-β1 protein levels and interfered with PI3K-AKT signal transduction pathway.
Conclusions
Our findings revealed that DHA could inhibit proliferation and migration of human hepatocellular carcinoma cells targeting FN1 and ITGB1 via the PI3K-AKT pathway. Therefore, DHA might be a novel drug with a potential effect against liver tumorigenesis and metastasis.
Similar content being viewed by others
Data availability
All data used to support the findings of this study are included within the article.
References
Aung W, Sogawa C, Furukawa T, Saga T (2011) Anticancer effect of dihydroartemisinin (DHA) in a pancreatic tumor model evaluated by conventional methods and optical imaging. Anticancer Res 31:1549–1558
Guo K et al (2008) Involvement of protein kinase C beta-extracellular signal-regulating kinase 1/2/p38 mitogen-activated protein kinase-heat shock protein 27 activation in hepatocellular carcinoma cell motility and invasion. Cancer Sci 99:486–496. https://doi.org/10.1111/j.1349-7006.2007.00702.x
He B et al (2016) hTERT mediates gastric cancer metastasis partially through the indirect targeting of ITGB1 by microRNA-29a. Sci Rep 6:1–12
Hlady RA et al (2018) Integrating the epigenome to identify novel drivers of hepatocellular carcinoma. Hepatology. https://doi.org/10.1002/hep.30211
Ho WE, Peh HY, Chan TK, Wong WS (2014) Artemisinins: pharmacological actions beyond anti-malarial. Pharmacol Ther 142:126–139. https://doi.org/10.1016/j.pharmthera.2013.12.001
Howe EN, Cochrane DR, Richer JK (2011) Targets of miR-200c mediate suppression of cell motility and anoikis resistance. Breast Cancer Res 13(2):R45. https://doi.org/10.1186/bcr2867
Kawahara R, Niwa Y, Simizu S (2018) Integrin β1 is an essential factor in vasculogenic mimicry of human cancer cells. Cancer Sci 109(8):2490–2496. https://doi.org/10.1111/cas.13693
Laudato S et al (2017) P53-induced miR-30e-5p inhibits colorectal cancer invasion and metastasis by targeting ITGA6 and ITGB1. Int J Cancer 141:1879–1890. https://doi.org/10.1002/ijc.30854
Li Y et al (2001) Establishment of cell clones with different metastatic potential from the metastatic hepatocellular carcinoma cell line MHCC97. World J Gastroenterol 7:630–636
Li Y et al (2004) Stepwise metastatic human hepatocellular carcinoma cell model system with multiple metastatic potentials established through consecutive in vivo selection and studies on metastatic characteristics. J Cancer Res Clin Oncol 130:460–468. https://doi.org/10.1007/s00432-004-0564-9
Liu F, Zhang J, Qian J, Wu G, Ma Z (2018) Emodin alleviates CCl4induced liver fibrosis by suppressing epithelialmesenchymal transition and transforming growth factorbeta1 in rats. Mol Med Rep 18:3262–3270. https://doi.org/10.3892/mmr.2018.9324
Shi F, Sottile J (2011) MT1-MMP regulates the turnover and endocytosis of extracellular matrix fibronectin. J Cell Sci 124(23):4039–4050. https://doi.org/10.1242/jcs.087858
Slezakova S, Ruda-Kucerova J (2017) Anticancer activity of artemisinin and its derivatives anticancer. Research 37:5995–6003. https://doi.org/10.21873/anticanres.12046
Sponziello M et al (2016) Fibronectin-1 expression is increased in aggressive thyroid cancer and favors the migration and invasion of cancer cells. Mol Cell Endocrinol 431:123–132
Teja-Isavadharm P et al (1996) Comparative bioavailability of oral, rectal, and intramuscular artemether in healthy subjects: use of simultaneous measurement by high performance liquid chromatography and bioassay. Br J Clin Pharmacol 42:599–604
Titulaer HA, Zuidema J, Kager PA, Wetsteyn JC, Lugt CB, Merkus FW (1990) The pharmacokinetics of artemisinin after oral, intramuscular and rectal administration to volunteers. J Pharm Pharmacol 42:810–813
Tong Y et al (2016) Artemisinin and its derivatives can significantly inhibit lung tumorigenesis and tumor metastasis through Wnt/beta-catenin signaling. Oncotarget 7:31413–31428. https://doi.org/10.18632/oncotarget.8920
Tu Y (1999) The development of new antimalarial drugs: qinghaosu and dihydro-qinghaosu. Chin Med J 112:976–977
Xu K, Al-Ani MK, Wang C, Qiu X, Chi Q, Zhu P, Dong N (2018a) Emodin as a selective proliferative inhibitor of vascular smooth muscle cells versus endothelial cells suppress arterial intima formation. Life Sci 207:9–14. https://doi.org/10.1016/j.lfs.2018.05.042
Xu K, Zhou T, Huang Y, Chi Q, Shi J, Zhu P, Dong N (2018b) Anthraquinone emodin inhibits tumor necrosis factor alpha-induced calcification of human aortic valve interstitial cells via the NF-kappaB pathway. Front Pharmacol 9:1328. https://doi.org/10.3389/fphar.2018.01328
Xu K, Sha Y, Wang S, Chi Q, Liu Y, Wang C, Yang L (2019) Effects of Bakuchiol on chondrocyte proliferation via the PI3K-Akt and ERK1/2 pathways mediated by the estrogen receptor for promotion of the regeneration of knee articular cartilage defects. Cell Prolif 52:e12666. https://doi.org/10.1111/cpr.12666
Yang X et al (2017) miR-200b regulates epithelial-mesenchymal transition of chemo-resistant breast cancer cells by targeting FN1. Discov Med 24:75–85
Zhang CZ, Zhang H, Yun J, Chen GG, Lai PB (2012) Dihydroartemisinin exhibits antitumor activity toward hepatocellular carcinoma in vitro and in vivo. Biochem Pharmacol 83:1278–1289. https://doi.org/10.1016/j.bcp.2012.02.002
Acknowledgements
This work was supported by National Natural Science Foundation of China (11832008, 11532004, 11902058) and Innovation and Attracting Talents Program for College and University (“111” Project) (Grant No. B06023).
Author information
Authors and Affiliations
Corresponding authors
Ethics declarations
Conflict of interest
All authors read and approved the final manuscript. We declare that the authors have no conflict of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Wu, R., Gao, Y., Wu, J. et al. Semi-synthetic product dihydroartemisinin inhibited fibronectin-1 and integrin-β1 and interfered with the migration of HCCLM6 cells via PI3K-AKT pathway. Biotechnol Lett 42, 917–926 (2020). https://doi.org/10.1007/s10529-020-02839-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10529-020-02839-8