Advances in Clinical and Experimental Medicine
2018, vol. 27, nr 7, July, p. 899–905
Publication type: original article
Inhibition of migration and invasion by berberine via inactivation of PI3K/Akt and p38 in human retinoblastoma cell line
1 Department of Ophthalmology, Ningbo Eye Hospital, China
Background. As a clinically important natural isoquinoline alkaloid, berberine has been reported to possess various pharmacological effects.
Objectives. This study was aimed to investigate the effect of berberine on cell migration and invasion in human retinoblastoma (Rb) cells.
Material and Methods. The cytotoxicity of berberine was estimated by 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) assay. After being stimulated with berberine under various concentrations, the cell migration and invasion were evaluated by transwell assay. Then, the expression levels of epithelial-mesenchymal transition (EMT) markers were determined by quantitative reverse transcription PCR (qRT-PCR) and western blot analysis. Furthermore, the phosphorylation levels of protein kinase B (Akt) and p38 were detected by western blot analysis. Finally, the effect of phosphatidylinositol-3-kinase (PI3K) and p38 inhibitors on cell migration and invasion was estimated by transwell assay. Untreated cells acted as control for all the experiments.
Results. The concentrations of berberine for further studies were controlled in a range of 0 to 100 μM. The cell migration and invasion were both suppressed by berberine in a dose-dependent manner compared to the control (p < 0.05 or p < 0.001). Berberine remarkably down-regulated expression of E-cadherin and up-regulated expression of vimentin and α-SMA compared to the control (p < 0.01 or p < 0.001). Furthermore, the phosphorylation levels of Akt and p38 were both down-regulated by berberine in comparison to the control. Furthermore, the addition of berberine accompanied by LY294002 or SB203580 significantly suppressed cell migration and invasion compared to the addition of berberine alone (p < 0.05).
Conclusion. Berberine suppressed cell migration and invasion via inactivation of PI3K/Akt and p38.
berberine, invasion, epithelial-mesenchymal transition, phosphatidylinositol 3-kinase/protein kinase B/Akt, p38
- Abramson DH, Schefler AC. Update on retinoblastoma. Retina. 2005;7:174–178.
- Friend SH, Bernards R, Rogelj S, et al. A human DNA segment with properties of the gene that predisposes to retinoblastoma and osteosarcoma. Nature. 1986;323:187–206.
- Parkin DM, Stiller CA, Draper GJ, Bieber CA. The international incidence of childhood cancer. Int J Cancer. 1988;42:511–520.
- Phan IT, Stout T. Retinoblastoma presenting as strabismus and leukocoria. J Pediatrics. 2010;157:858–858.
- Chawla B, Jain A, Azad R. Conservative treatment modalities in retinoblastoma. Ind J Ophthalmol. 2013;61:479–485.
- Pandey AN. Retinoblastoma: An overview. Saudi J Ophthalmol. 2014; 28:310–315.
- EI Kettani A, Aderdour S, Daghouj G, et al. Retinoblastoma: Preliminary results of national treatment protocol at Casablanca university medical center. J Fr Ophtalmol. 2014;37:115–124.
- Chang MW, Barr E, Seltzer J, et al. Cytostatic gene therapy for vascular proliferative disorders with a constitutively active form of the retinoblastoma gene product. Science. 1995;267:518–522.
- Durairajan S, Liu LF, Lu JH, Chen LL. Berberine ameliorates β-amyloid pathology, gliosis, and cognitive impairment in an Alzheimer’s disease transgenic mouse model. Neurobiol Aging. 2012;33:2903–2919.
- Gulbahar O, Ozturk G, Erdem N, Kazandi AC, Kokuludag A. Antioxidant and anti-inflammatory activities of berberine in the treatment of diabetes mellitus: Evidence-based complementary and alternative medicine. eCAM. 2014;2014:71–80.
- Chang W, Chen L, Hatch GM. Berberine as a therapy for type 2 diabetes and its complications: From mechanism of action to clinical studies. Biochem Cell Biol. 2014;1:1–8.
- Cicero AF, Ertek S. Metabolic and cardiovascular effects of berberine: From preclinical evidences to clinical trial results. Clin Lipidol. 2015;4:553–563.
- Xie J, Xu Y, Huang X, et al. Berberine-induced apoptosis in human breast cancer cells is mediated by reactive oxygen species generation and mitochondrial-related apoptotic pathway. Tumour Biol. 2015;36:1279–1288.
- Park JJ, Seo SM, Kim EJ, et al. Berberine inhibits human colon cancer cell migration via AMP-activated protein kinase-mediated downregulation of integrin β1 signaling. Biochem Biophys Res Commun. 2012;426:461–467.
- Chou HC, Lu YC, Cheng CS, et al. Proteomic and redox-proteomic analysis of berberine-induced cytotoxicity in breast cancer cells. J Proteomics. 2012;75:3158–3176.
- Yip NK, Ho WS. Berberine induces apoptosis via the mitochondrial pathway in liver cancer cells. Oncol Rep. 2013;30:1107–1112.
- Diogo CV, Machado NG, Barbosa IA, Serafim TL, Burgeiro A, Oliveira PJ. Berberine as a promising safe anti-cancer agent − is there a role for mitochondria? Curr Drug Targets. 2011;12:850–859.
- Chambers AF, Groom AC, Macdonald IC. Metastasis: Dissemination and growth of cancer cells in metastatic sites. Nat Rev Cancer. 2002;2: 563–572.
- Chen PN, Hsieh YS, Chiou HL, Chu SC. Silibinin inhibits cell invasion through inactivation of both PI3K-Akt and MAPK signaling pathways. Chem Biol Interact. 2005;156:141–150.
- Chen KS, Shi MD, Chien CS, Shih YW. Pinocembrin suppresses TGF-β1-induced epithelial-mesenchymal transition and metastasis of human Y-79 retinoblastoma cells through inactivating αvβ3 integrin/FAK/p38α signaling pathway. Cell Biosci. 2014;4:1–13.
- Schefe JH, Lehmann KE, Buschmann IR, et al. Quantitative real-time RT-PCR data analysis: Current concepts and the novel ‘gene expression’s CT difference’ formula. J Mol Med. 2006;84:901–910.
- Ho YT, Yang JS, Li TC, et al. Berberine suppresses in vitro migration and invasion of human SCC-4 tongue squamous cancer cells through the inhibitions of FAK, IKK, NF-κB, u-PA and MMP-2 and -9. Cancer Lett. 2009;279:155–162.
- Yan L, Yan K, Kun W, et al. Berberine inhibits the migration and invasion of T24 bladder cancer cells via reducing the expression of heparanase. Tumour Biol. 2013;34:215–221.
- Liu X, Ji Q, Ye N, et al. Berberine inhibits invasion and metastasis of colorectal cancer cells via COX-2/PGE2 mediated JAK2/STAT3 signaling pathway. PLOS One. 2015;10:e0123478.
- Qi H, Xin L, Xu X, Ji X, Fan L. Epithelial-to-mesenchymal transition markers to predict response of berberine in suppressing lung cancer invasion and metastasis. J Transl Med. 2014;12:116–122.
- Medici D, Muñoz-Cánoves P, Yang PC, Brunelli S. Mesenchymal transitions in development and disease. Stem Cells Int. 2016;2016:1–2.
- Zong H, Yin B, Zhou H, Cai D, Ma B, Xiang Y. Inhibition of mTOR pathway attenuates migration and invasion of gallbladder cancer via EMT inhibition. Mol Biol Rep. 2014;41:4507–4512.
- Li S, Qin X, Li Y, et al. MiR-133a suppresses the migration and invasion of esophageal cancer cells by targeting the EMT regulator SOX4. Am J Transl Res. 2015;7:1390–1403.
- Villegas-Comonfort S, Castillo-Sanchez R, Serna-Marquez N, Cortes-Reynosa P, Salazar EP. Arachidonic acid promotes migration and invasion through a PI3K/Akt-dependent pathway in MDA-MB-231 breast cancer cells. Prostaglandins Leukot Essent Fatty Acids. 2014;90:169–177.
- Coulthard LR, White DE, Jones DL, Mcdermott MF, Burchill SA. p38 MAPK: Stress responses from molecular mechanisms to therapeutics. Trends Mol Med. 2009;15:369–379.
- Cheng GZ, Chan J, Wang Q, Zhang W, Sun CD, Wang LH. Twist transcriptionally up-regulates AKT2 in breast cancer cells leading to increased migration, invasion, and resistance to paclitaxel. Cancer Res. 2007;67:1979–1987.
- Ma XM, Shen ZH, Liu ZY, et al. Heparanase promotes human gastric cancer cells migration and invasion by increasing Src and p38 phosphorylation expression. Int J Clin Exp Pathol. 2014;7:5609–5621.
- Liu Y, Zheng J, Zhang Y, et al. Fucoxanthin activates apoptosis via inhibition of pi3k/akt/mtor pathway and suppresses invasion and migration by restriction of p38-MMP-2/9 pathway in human glioblastoma cells. Neurochem Res. 2016;41:2728–2751.
- Li F, Yin X, Luo X, et al. Livin promotes progression of breast cancer through induction of epithelial–mesenchymal transition and activation of AKT signaling. Cell Signal. 2013;25:1413–1422.
- Liu L, Dai Y, Chen J, et al. Maelstrom promotes hepatocellular carcinoma metastasis by inducing epithelial‐mesenchymal transition by way of Akt/GSK‐3β/snail signaling. Hepatology. 2014;59:531–543.
- Kolosova I, Nethery D, Kern JA. Role of Smad2/3 and p38 MAP kinase in TGF-β1-induced epithelial–mesenchymal transition of pulmonary epithelial cells. J Cell Physiol. 2011;226:1248–1254.