Tanshinone IIA reverses EGF‑ and TGF‑β1‑mediated epithelial‑mesenchymal transition in HepG2 cells via the PI3K/Akt/ERK signaling pathway

Zhang,Longkai; Lin,Weibin; Chen,Xiaodan; Wei,Gang; Zhu,Hailong; Xing,Shangping

doi:10.3892/ol.2019.11032

Tanshinone IIA reverses EGF‑ and TGF‑β1‑mediated epithelial‑mesenchymal transition in HepG2 cells via the PI3K/Akt/ERK signaling pathway

Authors:
- Longkai Zhang
- Weibin Lin
- Xiaodan Chen
- Gang Wei
- Hailong Zhu
- Shangping Xing
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Affiliations: Traditional Chinese Medicine Quality Evaluation and Testing Center, Hong Zheng Dao (China) Traditional Chinese Medicine Research Company Ltd., Guangzhou, Guangdong 510006, P.R. China, School of Pharmaceutical Science, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 510006, P.R. China
Published online on: November 1, 2019 https://doi.org/10.3892/ol.2019.11032
Pages: 6554-6562
Copyright: © Zhang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Epithelial‑to‑mesenchymal transition (EMT) is an essential phenotypic conversion involved in cancer progression. Epidermal growth factor (EGF) and transforming growth factor (TGF)‑β1 are potent inducers of the EMT. Tanshinone IIA (Tan IIA) is a phenanthrenequinone extracted from the root of Salvia miltiorrhiza Bunge, and its anticancer activity has been demonstrated in numerous studies. However, the mechanisms of action underlying Tan IIA in EGF‑ and TGF‑β1‑induced EMT in HepG2 cells remain unknown. Multiple assays were utilized in the present study, including colony formation, wound healing, Transwell invasion, immunoﬂuorescence staining and western blotting, in order to assess the inﬂuence of Tan IIA on HepG2 cells induced by 20 ng/ml EGF and 10 ng/ml TGF‑β1. The present study reported that Tan IIA treatment decreased EGF‑ and TGF‑β1‑enhanced cell colony numbers, migration and invasion, and inhibited EGF‑ and TGF‑β1‑induced decreases in the expression levels of E‑cadherin, and increases in the expression levels of matrix metalloproteinase‑2, N‑cadherin, vimentin and Snail. In addition, it was observed that Tan IIA decreased the expression levels of phosphorylated (p)‑Akt and p‑ERK1/2 induced by EGF and TGF‑β1. Furthermore, western blot analysis confirmed that blocking the function of PI3K/Akt and ERK with LY294002 and U0126 resulted in upregulation of E‑cadherin expression, and downregulation of vimentin and Snail expression in EGF‑ and TGF‑β1‑treated HepG2 cells. In conclusion, to the best of our knowledge, the results of the present study are the first to indicate that Tan IIA may suppress EGF‑ and TGF‑β1‑induced EMT in HepG2 cells by deactivating the PI3K/Akt/ERK pathway.

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1	Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA and Jemal A: Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 68:394–424. 2018. View Article : Google Scholar : PubMed/NCBI
2	Mittal V: Epithelial mesenchymal transition in tumor metastasis. Annu Rev Pathol. 13:395–412. 2018. View Article : Google Scholar : PubMed/NCBI
3	Mehlen P and Puisieux A: Metastasis: A question of life or death. Nat Rev Cancer. 6:449–458. 2006. View Article : Google Scholar : PubMed/NCBI
4	Pastushenko I, Brisebarre A, Sifrim A, Fioramonti M, Revenco T, Boumahdi S, Van Keymeulen A, Brown D, Moers V, Lemaire S, et al: Identification of the tumour transition states occurring during EMT. Nature. 556:463–468. 2018. View Article : Google Scholar : PubMed/NCBI
5	Puisieux A, Brabletz T and Caramel J: Oncogenic roles of EMT-inducing transcription factors. Nat Cell Biol. 16:488–494. 2014. View Article : Google Scholar : PubMed/NCBI
6	Said NA and Williams ED: Growth factors in induction of epithelial-mesenchymal transition and metastasis. Cells Tissues Organs. 193:85–97. 2011. View Article : Google Scholar : PubMed/NCBI
7	Xing S, Yu W, Zhang X, Luo Y, Lei Z, Huang D, Lin J, Huang Y, Huang S, Nong F, et al: Isoviolanthin extracted from dendrobium officinale reverses TGF-β1-mediated epithelial-mesenchymal transition in hepatocellular carcinoma cells via deactivating the TGF-β/Smad and PI3K/Akt/mTOR signaling pathways. Int J Mol Sci. 19(pii): E15562018. View Article : Google Scholar : PubMed/NCBI
8	Fantozzi A, Gruber DC, Pisarsky L, Heck C, Kunita A, Yilmaz M, Meyer-Schaller N, Cornille K, Hopfer U, Bentires-Alj M and Christofori G: VEGF-mediated angiogenesis links EMT-induced cancer stemness to tumor initiation. Cancer Res. 74:1566–1575. 2014. View Article : Google Scholar : PubMed/NCBI
9	Mori S, Kodaira M, Ito A, Okazaki M, Kawaguchi N, Hamada Y, Takada Y and Matsuura N: Enhanced expression of integrin αvβ3 induced by TGF-β Is required for the enhancing effect of fibroblast growth factor 1 (FGF1) in TGF-β-induced epithelial-mesenchymal transition (EMT) in mammary epithelial cells. PLoS One. 10:e01374862015. View Article : Google Scholar : PubMed/NCBI
10	Grassi ML, Palma CS, Thome CH, Lanfredi GP, Poersch A and Faca VM: Proteomic analysis of ovarian cancer cells during epithelial-mesenchymal transition (EMT) induced by epidermal growth factor (EGF) reveals mechanisms of cell cycle control. J Proteomics. 151:2–11. 2017. View Article : Google Scholar : PubMed/NCBI
11	Stewart TA, Azimi I, Brooks AJ, Thompson EW, Roberts-Thomson SJ and Monteith GR: Janus kinases and Src family kinases in the regulation of EGF-induced vimentin expression in MDA-MB-468 breast cancer cells. Int J Biochem Cell Biol. 76:64–74. 2016. View Article : Google Scholar : PubMed/NCBI
12	Miyazono K: Transforming growth factor-beta signaling in epithelial-mesenchymal transition and progression of cancer. Proc Jpn Acad Ser B Phys Biol Sci. 85:314–323. 2009. View Article : Google Scholar : PubMed/NCBI
13	Ren W, Zhang Y, Zhang L, Lin Q, Zhang J and Xu G: Overexpression of collagen type V α1 chain in human breast invasive ductal carcinoma is mediated by TGF-β1. Int J Oncol. Mar 15–2018.(Epub ahead of print). View Article : Google Scholar
14	Li ZM, Xu SW and Liu PQ: Salvia miltiorrhiza Burge (Danshen): A golden herbal medicine in cardiovascular therapeutics. Acta Pharmacol Sin. 39:802–824. 2018. View Article : Google Scholar : PubMed/NCBI
15	Huang L, Zhu J, Zheng M, Zou R, Zhou Y and Zhu M: Tanshinone IIA protects against subclinical lipopolysaccharide induced cardiac fibrosis in mice through inhibition of NADPH oxidase. Int Immunopharmacol. 60:59–63. 2018. View Article : Google Scholar : PubMed/NCBI
16	Chen W, Tang F, Xie B, Chen S, Huang H and Liu P: Amelioration of atherosclerosis by tanshinone IIA in hyperlipidemic rabbits through attenuation of oxidative stress. Eur J Pharmacol. 674:359–364. 2012. View Article : Google Scholar : PubMed/NCBI
17	Gao S, Liu Z, Li H, Little PJ, Liu P and Xu S: Cardiovascular actions and therapeutic potential of tanshinone IIA. Atherosclerosis. 220:3–10. 2012. View Article : Google Scholar : PubMed/NCBI
18	Lu BL, Li J, Zhou J, Li WW and Wu HF: Tanshinone IIA decreases the levels of inflammation induced by Aβ1–42 in brain tissues of Alzheimer's disease model rats. Neuroreport. 27:883–893. 2016. View Article : Google Scholar : PubMed/NCBI
19	Cao YF, Wang SF, Li X, Zhang YL and Qiao YJ: The anticancer mechanism investigation of Tanshinone II_A by pharmacological clustering in protein network. BMC Syst Biol. 12:902018. View Article : Google Scholar : PubMed/NCBI
20	Zhu YQ, Wang BY, Wu F, An YK and Zhou XQ: Influence of tanshinone IIA on the apoptosis of human esophageal Ec-109 cells. Nat Prod Commun. 11:17–19. 2016.PubMed/NCBI
21	He L and Gu K: Tanshinone IIA regulates colorectal cancer apoptosis via attenuation of Parkin-mediated mitophagy by suppressing AMPK/Skp2 pathways. Mol Med Rep. 18:1692–1703. 2018.PubMed/NCBI
22	Su CC, Chien SY, Kuo SJ, Chen YL, Cheng CY and Chen DR: Tanshinone IIA inhibits human breast cancer MDA-MB-231 cells by decreasing LC3-II, Erb-B2 and NF-κBp65. Mol Med Rep. 5:1019–1022. 2012. View Article : Google Scholar : PubMed/NCBI
23	Kim EO, Kang SE, Im CR, Lee JH, Ahn KS, Yang WM, Um JY, Lee SG and Yun M: Tanshinone IIA induces TRAIL sensitization of human lung cancer cells through selective ER stress induction. Int J Oncol. 48:2205–2212. 2016. View Article : Google Scholar : PubMed/NCBI
24	Lin CY, Chang TW, Hsieh WH, Hung MC, Lin IH, Lai SC and Tzeng YJ: Simultaneous induction of apoptosis and necroptosis by Tanshinone IIA in human hepatocellular carcinoma HepG2 cells. Cell Death Discov. 2:160652016. View Article : Google Scholar : PubMed/NCBI
25	Qin J, Shi H, Xu Y, Zhao F and Wang Q: Tanshinone IIA inhibits cervix carcinoma stem cells migration and invasion via inhibiting YAP transcriptional activity. Biomed Pharmacother. 105:758–765. 2018. View Article : Google Scholar : PubMed/NCBI
26	Huang SY, Chang SF, Liao KF and Chiu SC: Tanshinone IIA inhibits epithelial-mesenchymal transition in bladder cancer cells via modulation of STAT3-CCL2 signaling. Int J Mol Sci. 18(pii): E16162017. View Article : Google Scholar : PubMed/NCBI
27	Zhou M, Zhou G, Hu S and Zhang L: Tanshinone IIA suppress the proliferation of HNE-1 nasopharyngeal carcinoma an in vitro study. Saudi J Biol Sci. 25:267–272. 2018. View Article : Google Scholar : PubMed/NCBI
28	Fu H, He Y, Qi L, Chen L, Luo Y, Chen L, Li Y, Zhang N and Guo H: cPLA2α activates PI3K/AKT and inhibits Smad2/3 during epithelial-mesenchymal transition of hepatocellular carcinoma cells. Cancer Lett. 403:260–270. 2017. View Article : Google Scholar : PubMed/NCBI
29	Xing S, Zhang X, Ke H, Lin J, Huang Y and Wei G: Physicochemical properties of polysaccharides from Dendrobium officinale by fractional precipitation and their preliminary antioxidant and anti-HepG2 cells activities in vitro. Chem Cent J. 12:1002018. View Article : Google Scholar : PubMed/NCBI
30	Cheng Z, Wei W, Wu Z, Wang J, Ding X, Sheng Y, Han Y and Wu Q: ARPC2 promotes breast cancer proliferation and metastasis. Oncol Rep. 41:3189–3200. 2019.PubMed/NCBI
31	Deng G, Zeng S, Ma J, Zhang Y, Qu Y, Han Y, Yin L, Cai C, Guo C and Shen H: The anti-tumor activities of Neferine on cell invasion and oxaliplatin sensitivity regulated by EMT via Snail signaling in hepatocellular carcinoma. Sci Rep. 7:416162017. View Article : Google Scholar : PubMed/NCBI
32	Gore J, Deitz SL, Wilson JL and Korc M: Abstract 969: TGF-beta cross-talks with the EGF receptor family to promote proliferation of pancreatic cancer cells with dysfunctional RB. Cancer Res. 74:9692014.
33	Xu ZH, Jiang YY, Steed H, Davidge S and Fu YX: TGFβ and EGF synergistically induce a more invasive phenotype of epithelial ovarian cancer cells. Biochem Biophys Res Commun. 401:376–381. 2010. View Article : Google Scholar : PubMed/NCBI
34	Wendt MK, Allington TM and Schiemann WP: Mechanisms of the epithelial-mesenchymal transition by TGF-beta. Future Oncol. 5:1145–1168. 2009. View Article : Google Scholar : PubMed/NCBI
35	Lu ZM, Ghosh S, Wang ZY and Hunter T: Downregulation of caveolin-1 function by EGF leads to the loss of E-cadherin, increased transcriptional activity of beta-catenin, and enhanced tumor cell invasion. Cancer Cell. 4:499–515. 2003. View Article : Google Scholar : PubMed/NCBI
36	Xu S and Liu P: Tanshinone II-A: New perspectives for old remedies. Expert Opin Ther Pat. 23:149–153. 2013. View Article : Google Scholar : PubMed/NCBI
37	Jeon YJ, Kim JS, Hwang GH, Wu Z, Han HJ, Park SH, Chang W, Kim LK, Lee YM, Liu KH and Lee MY: Inhibition of cytochrome P450 2J2 by tanshinone IIA induces apoptotic cell death in hepatocellular carcinoma HepG2 cells. Eur J Pharmacol. 764:480–488. 2015. View Article : Google Scholar : PubMed/NCBI
38	Ren X, Wang C, Xie B, Hu L, Chai H, Ding L, Tang L, Xia Y and Dou X: Tanshinone IIA induced cell death via miR30b-p53-PTPN11/SHP2 signaling pathway in human hepatocellular carcinoma cells. Eur J Pharmacol. 796:233–241. 2017. View Article : Google Scholar : PubMed/NCBI
39	Dong W, Zhang Y, Chen X and Jia Y: High-dose tanshinone iia suppresses migration and proliferation while promoting apoptosis of astrocytoma cells via Notch-1 pathway. Neurochem Res. 43:1855–1861. 2018. View Article : Google Scholar : PubMed/NCBI
40	Duan H, Ma L, Liu H, Zhang Y, Zhang Z, Yan X and Li X: Tanshinone IIA attenuates epithelial-mesenchymal transition to inhibit the tracheal narrowing. J Surg Res. 206:252–262. 2016. View Article : Google Scholar : PubMed/NCBI
41	Pan M, Schinke H, Luxenburger E, Kranz G, Shakhtour J, Libl D, Huang Y, Gaber A, Pavšič M, Lenarčič B, et al: EpCAM ectodomain EpEX is a ligand of EGFR that counteracts EGF-mediated epithelial-mesenchymal transition through modulation of phospho-ERK1/2 in head and neck cancers. PLoS Biol. 16:e20066242018. View Article : Google Scholar : PubMed/NCBI
42	Sheng W, Chen C, Dong M, Wang G, Zhou J, Song H, Li Y, Zhang J and Ding S: Calreticulin promotes EGF-induced EMT in pancreatic cancer cells via Integrin/EGFR-ERK/MAPK signaling pathway. Cell Death Dis. 8:e31472017. View Article : Google Scholar : PubMed/NCBI
43	Bhat FA, Sharmila G, Balakrishnan S, Arunkumar R, Elumalai P, Suganya S, Raja Singh P, Srinivasan N and Arunakaran J: Quercetin reverses EGF-induced epithelial to mesenchymal transition and invasiveness in prostate cancer (PC-3) cell line via EGFR/PI3K/Akt pathway. J Nutr Biochem. 25:1132–1139. 2014. View Article : Google Scholar : PubMed/NCBI
44	Tsai PC, Fu YS, Chang LS and Lin SR: Taiwan cobra cardiotoxin III suppresses EGF/EGFR-mediated epithelial-to-mesenchymal transition and invasion of human breast cancer MDA-MB-231 cells. Toxicon. 111:108–120. 2016. View Article : Google Scholar : PubMed/NCBI
45	Liu Z, Kuang W, Zhou Q and Zhang Y: TGF-β1 secreted by M2 phenotype macrophages enhances the stemness and migration of glioma cells via the SMAD2/3 signalling pathway. Int J Mol Med. 42:3395–3403. 2018.PubMed/NCBI
46	Ouanouki A, Lamy S and Annabi B: Anthocyanidins inhibit epithelial-mesenchymal transition through a TGFβ/Smad2 signaling pathway in glioblastoma cells. Mol Carcinog. 56:1088–1099. 2017. View Article : Google Scholar : PubMed/NCBI
47	Pei Z, Fu W and Wang G: A natural product toosendanin inhibits epithelial-mesenchymal transition and tumor growth in pancreatic cancer via deactivating Akt/mTOR signaling. Biochem Biophys Res Commun. 493:455–460. 2017. View Article : Google Scholar : PubMed/NCBI
48	Han S, Bui NT, Ho MT, Kim YM, Cho M and Shin DB: Dexamethasone inhibits TGF-β1-induced cell migration by regulating the ERK and AKT pathways in human colon cancer cells Via CYR61. Cancer Res Treat. 48:1141–1153. 2016. View Article : Google Scholar : PubMed/NCBI
49	Fuxe J, Vincent T and Garcia de Herreros A: Transcriptional crosstalk between TGF-β and stem cell pathways in tumor cell invasion: Role of EMT promoting Smad complexes. Cell Cycle. 9:2363–2374. 2010. View Article : Google Scholar : PubMed/NCBI