Emodin, a natural anthraquinone, suppresses liver cancer in vitro and in vivo by regulating VEGFR2 and miR-34a
- 30 Downloads
The pharmacokinetic (PK) and potential effects of Emodin on liver cancer were systematically evaluated in this study. Both the intragastric administration (i.g.) and hypodermic injection (i.h.) of Emodin exhibited a strong absorption (absorption rate < 1 h) and elimination capacity (t1/2 ≈ 2 h). The tissue distribution of Emodin after i.h. was rapid and wide. The stability of Emodin in three species of liver microsomes wasrat >human> beagle dog. These PK data provided the basis for the subsequent animal experiments. In liver cancer patient tissues, the expression of vascular endothelial growth factor (VEGF)-induced signaling pathways, including phosphorylated VEGF receptor 2 (VEGFR2), AKT, and ERK1/2,were simultaneously elevated, but miR-34a expression was reduced and negatively correlated with SMAD2 and SMAD4. Emodin inhibited the expression of SMAD2/4 in HepG2 cells by inducing the miR-34a level. Subsequently, BALB/c nude mice received a daily subcutaneous injection of HepG2 cells with or without Emodin treatment (1 mg/kg or 10 mg/kg), and Emodin inhibited tumorigenesis and reduced the mortality rate in a dose-dependent manner. In vivo experiments showed that cell proliferation, migration, and invasion were promoted by VEGF or miR-34a signal treatment but were inhibited when combined with Emodin treatment. All these results demonstrated that Emodin inhibited tumorigenesis in liver cancer by simultaneously inhibiting the VEGFR2-AKT-ERK1/2signaling pathway and promoting a miR-34a-mediated signaling pathway.
KeywordsEmodin Liver cancer VEGFR2 miR-34a
R.T. and J. B. designed the experiments; C.S. performed the animal experiments; J. J. performed the cell experiments in vitro; Z. Y. and W. Y. performed the western blot analysis and the luciferase assay; G. M. and J. W. performed the immunohistochemistry assay; and J. B. analyzed the data and drafted the manuscript.
This work was supported by funding from the Fourth Hospital of Hebei Medical University.
Compliance with ethical standards
Conflict of interest
All authors have declared that they have no conflicts of interest to disclose. Jianguo Bai declares that he has no conflict of interest. Jianfei Wu declares that he has no conflict of interest. Ruifeng Tang declares that he has no conflict of interest. Chao Sun declares that he has no conflict of interest. Junwei Ji declares that he has no conflict of interest. Zhaolin Yin declares that he has no conflict of interest. Guangjun Ma declares that he has no conflict of interest. Wei Yang declares that he has no conflict of interest.
The Guide for the Care and Use of Laboratory Animals [National Research Council (US) Committee for the update of the Guide for the Care and Use of Laboratory Animals, 2011] was followed. All procedures in studies involving animals were performed in accordance with the ethical standards of the Animal Ethics Committee of the Fourth Hospital of Hebei Medical University (Shijiazhuang, Hebei Province, China).
For this type of study, formal consent is not required.
- 1.Adult Primary Liver Cancer Treatment (PDQ(R)) (2002) Patient version. PDQ Cancer Information Summaries. Bethesda (MD), InGoogle Scholar
- 3.Global, regional, and national life expectancy, all-cause mortality, and cause-specific mortality for 249 causes of death, 1980–2015: a systematic analysis for the Global Burden of Disease Study 2015 (2016). Lancet 388 (10053):1459–1544. https://doi.org/10.1016/S0140-6736(16)31012-1
- 4.Global, regional, and national age-sex specific all-cause and cause-specific mortality for 240 causes of death, 1990–2013: a systematic analysis for the Global Burden of Disease Study 2013 (2015). Lancet 385 (9963):117–171. https://doi.org/10.1016/S0140-6736(14)61682-2
- 6.Wang Z, Zhang G, Wu J, Jia M (2013) Adjuvant therapy for hepatocellular carcinoma: current situation and prospect. Drug Discov Ther 7(4):137–143Google Scholar
- 8.Chang LC, Huang N, Chou YJ, Lee CH, Kao FY, Huang YT (2008) Utilization patterns of Chinese medicine and Western medicine under the National Health Insurance Program in Taiwan, a population-based study from 1997 to 2003. BMC Health Serv Res 8:170. https://doi.org/10.1186/1472-6963-8-170 Google Scholar
- 12.Zou Y, Guo CG, Yang ZG, Sun JH, Zhang MM, Fu CY (2016) A small interfering RNA targeting vascular endothelial growth factor efficiently inhibits growth of VX2 cells and VX2 tumor model of hepatocellular carcinoma in rabbit by transarterial embolization-mediated siRNA delivery. Drug Des Devel Ther 10:1243–1255. https://doi.org/10.2147/DDDT.S94122 Google Scholar
- 14.Lin CC, Ng LT, Hsu FF, Shieh DE, Chiang LC (2004) Cytotoxic effects of Coptis chinensis and Epimedium sagittatum extracts and their major constituents (berberine, coptisine and icariin) on hepatoma and leukaemia cell growth. Clin Exp Pharmacol Physiol 31(1–2):65–69Google Scholar
- 15.Chang WC, Lin YL, Lee MJ, Shiow SJ, Wang CJ (1996) Inhibitory effect of crocetin on benzo(a)pyrene genotoxicity and neoplastic transformation in C3H10T1/2 cells. Anticancer Res 16(6B):3603–3608Google Scholar
- 16.Wang JB, Zhao HP, Zhao YL, Jin C, Liu DJ, Kong WJ, Fang F, Zhang L, Wang HJ, Xiao XH (2011) Hepatotoxicity or hepatoprotection? Pattern recognition for the paradoxical effect of the Chinese herb Rheum palmatum L. in treating rat liver injury. PLoS One 6(9):e24498. https://doi.org/10.1371/journal.pone.0024498 Google Scholar
- 21.Kim J, Lee JS, Jung J, Lim I, Lee JY, Park MJ (2015) Emodin suppresses maintenance of stemness by augmenting proteosomal degradation of epidermal growth factor receptor/epidermal growth factor receptor variant III in glioma stem cells. Stem Cells Dev 24(3):284–295. https://doi.org/10.1089/scd.2014.0210 Google Scholar
- 24.Sun YP, Liu JP (2015) Blockade of emodin on amyloid-beta 25-35-induced neurotoxicity in AbetaPP/PS1 mice and PC12 cells through activation of the class III phosphatidylinositol 3-kinase/Beclin-1/B-cell lymphoma 2 pathway. Planta Med 81(2):108–115. https://doi.org/10.1055/s-0034-1383410 Google Scholar
- 30.Xie MJ, Ma YH, Miao L, Wang Y, Wang HZ, Xing YY, Xi T, Lu YY (2014) Emodin-provoked oxidative stress induces apoptosis in human colon cancer HCT116 cells through a p53-mitochondrial apoptotic pathway. Asian Pac J Cancer P 15(13):5201–5205Google Scholar
- 31.Kirsch M, Schackert G, Black PM (2004) Metastasis and angiogenesis. Cancer Treat Res 117:285–304Google Scholar
- 34.Torimura T, Sata M, Ueno T, Kin M, Tsuji R, Suzaku K, Hashimoto O, Sugawara H, Tanikawa K (1998) Increased expression of vascular endothelial growth factor is associated with tumor progression in hepatocellular carcinoma. Hum Pathol 29(9):986–991Google Scholar
- 35.Sternberg CN, Davis ID, Mardiak J, Szczylik C, Lee E, Wagstaff J, Barrios CH, Salman P, Gladkov OA, Kavina A, Zarba JJ, Chen M, McCann L, Pandite L, Roychowdhury DF, Hawkins RE (2010) Pazopanib in locally advanced or metastatic renal cell carcinoma: results of a randomized phase III trial. J Clin Oncol Off J Am Soc Clin Oncol 28(6):1061–1068. https://doi.org/10.1200/JCO.2009.23.9764 Google Scholar
- 36.Grothey A, Van Cutsem E, Sobrero A, Siena S, Falcone A, Ychou M, Humblet Y, Bouche O, Mineur L, Barone C, Adenis A, Tabernero J, Yoshino T, Lenz HJ, Goldberg RM, Sargent DJ, Cihon F, Cupit L, Wagner A, Laurent D (2013) Regorafenib monotherapy for previously treated metastatic colorectal cancer (CORRECT): an international, multicentre, randomised, placebo-controlled, phase 3 trial. Lancet 381(9863):303–312. https://doi.org/10.1016/S0140-6736(12)61900-X Google Scholar
- 38.Bartel DP (2004) MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116(2):281–297Google Scholar
- 42.Mraz M, Malinova K, Kotaskova J, Pavlova S, Tichy B, Malcikova J, Stano Kozubik K, Smardova J, Brychtova Y, Doubek M, Trbusek M, Mayer J, Pospisilova S (2009) miR-34a, miR-29c and miR-17-5p are downregulated in CLL patients with TP53 abnormalities. Leukemia 23(6):1159–1163. https://doi.org/10.1038/leu.2008.377 Google Scholar
- 47.Kwak HJ, Park MJ, Park CM, Moon SI, Yoo DH, Lee HC, Lee SH, Kim MS, Lee HW, Shin WS, Park IC, Rhee CH, Hong SI (2006) Emodin inhibits vascular endothelial growth factor-A-induced angiogenesis by blocking receptor-2 (KDR/Flk-1) phosphorylation. Int J Cancer 118(11):2711–2720. https://doi.org/10.1002/ijc.21641 Google Scholar
- 51.Jia X, Yu F, Wang J, Iwanowycz S, Saaoud F, Wang Y, Hu J, Wang Q, Fan D (2014) Emodin suppresses pulmonary metastasis of breast cancer accompanied with decreased macrophage recruitment and M2 polarization in the lungs. Breast Cancer Res Treat 148(2):291–302. https://doi.org/10.1007/s10549-014-3164-7 Google Scholar
- 56.Subramaniam A, Loo SY, Rajendran P, Manu KA, Perumal E, Li F, Shanmugam MK, Siveen KS, Park JI, Ahn KS, Hui KM, Kumar AP, Sethi G (2013) An anthraquinone derivative, emodin sensitizes hepatocellular carcinoma cells to TRAIL induced apoptosis through the induction of death receptors and downregulation of cell survival proteins. Apoptosis 18(10):1175–1187. https://doi.org/10.1007/s10495-013-0851-5 Google Scholar
- 57.Meijerink MR, Puijk RS, van Tilborg A, Henningsen KH, Fernandez LG, Neyt M, Heymans J, Frankema JS, de Jong KP, Richel DJ, Prevoo W, Vlayen J (2018) Radiofrequency and microwave ablation compared to systemic chemotherapy and to partial hepatectomy in the treatment of colorectal liver metastases: a systematic review and meta-analysis. Cardiovasc Intervent Radiol 41(8):1189–1204. https://doi.org/10.1007/s00270-018-1959-3 Google Scholar
- 61.Miura H, Miyazaki T, Kuroda M, Oka T, Machinami R, Kodama T, Shibuya M, Makuuchi M, Yazaki Y, Ohnishi S (1997) Increased expression of vascular endothelial growth factor in human hepatocellular carcinoma. J Hepatol 27(5):854–861Google Scholar
- 64.Hiyoshi Y, Schetter AJ, Okayama H, Inamura K, Anami K, Nguyen GH, Horikawa I, Hawkes JE, Bowman ED, Leung SY, Harris CC (2015) Increased microRNA-34b and -34c predominantly expressed in stromal tissues is associated with poor prognosis in human colon cancer. PLoS One 10(4):e0124899. https://doi.org/10.1371/journal.pone.0124899 Google Scholar
- 65.Jamieson NB, Morran DC, Morton JP, Ali A, Dickson EJ, Carter CR, Sansom OJ, Evans TR, McKay CJ, Oien KA (2012) MicroRNA molecular profiles associated with diagnosis, clinicopathologic criteria, and overall survival in patients with resectable pancreatic ductal adenocarcinoma. Clinical cancer research : an official journal of the American association for. Cancer Res 18(2):534–545. https://doi.org/10.1158/1078-0432.CCR-11-0679 Google Scholar
- 68.Cortez MA, Valdecanas D, Niknam S, Peltier HJ, Diao L, Giri U, Komaki R, Calin GA, Gomez DR, Chang JY, Heymach JV, Bader AG, Welsh JW (2015) In vivo delivery of miR-34a sensitizes lung tumors to radiation through RAD51 regulation. Mol Ther-Nucl Acids 4:e270. https://doi.org/10.1038/mtna.2015.47 Google Scholar
- 69.Bruna A, Darken RS, Rojo F, Ocana A, Penuelas S, Arias A, Paris R, Tortosa A, Mora J, Baselga J, Seoane J (2007) High TGFbeta-Smad activity confers poor prognosis in glioma patients and promotes cell proliferation depending on the methylation of the PDGF-B gene. Cancer Cell 11(2):147–160. https://doi.org/10.1016/j.ccr.2006.11.023 Google Scholar
- 72.Cortez MA, Ivan C, Valdecanas D, Wang X, Peltier HJ, Ye Y, Araujo L, Carbone DP, Shilo K, Giri DK, Kelnar K, Martin D, Komaki R, Gomez DR, Krishnan S, Calin GA, Bader AG, Welsh JW (2016) PDL1 regulation by p53 via miR-34. J Natl Cancer Inst 108(1). https://doi.org/10.1093/jnci/djv303