Puerarin inhibits M2 polarization and metastasis of tumor-associated macrophages from NSCLC xenograft model via inactivating MEK/ERK 1/2 pathway

Kang,Honggang; Zhang,Jing; Wang,Baozhong; Liu,Meirong; Zhao,Jun; Yang,Mengxiang; Li,Yingxue

doi:10.3892/ijo.2017.3841

Puerarin inhibits M2 polarization and metastasis of tumor-associated macrophages from NSCLC xenograft model via inactivating MEK/ERK 1/2 pathway

Authors:
- Honggang Kang
- Jing Zhang
- Baozhong Wang
- Meirong Liu
- Jun Zhao
- Mengxiang Yang
- Yingxue Li
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Affiliations: Department of Oncology, Liaocheng People's Hospital, Liaocheng, Shandong 252004, P.R. China, Department of Pathology, Liaocheng People's Hospital, Liaocheng, Shandong 252004, P.R. China
Published online on: January 5, 2017 https://doi.org/10.3892/ijo.2017.3841
Pages: 545-554

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Abstract

Non-small cell lung carcinoma (NSCLC) metastasis is responsible for most of cancer-related mortality. The tumor associated macrophages (TAMs) are known to be crucial cells in lung cancer and are usually divided into two antagonistic types, M1 and M2. Puerarin has a wide spectrum of pharmacological properties. The present study explores puerarin on macrophage polarization and metastasis of NSCLC. The results demonstrated that puerarin inhibited tumor growth and tumor volumes in NSCLC xenograft model, increased M1 markers [CD197+, inducible nitric oxide synthase (iNOS)+, CD40+)] and reduced M2 markers (CD206+, Arg-1+ and CD163+). Besides, puerarin elevated the level of pro-inflammatory cytokine interferon (IFN)-γ, tumor necrosis factor (TNF)-α and interleukin (IL)-12, decreased the expression of pro-tumor cytokines IL-10, IL-4 and transforming growth factor (TGF)-β. To explore whether puerarin directly acts on macrophages, we purified macrophages from NSCLC model, the results showed that puerarin inhibited macrophages polarized to M2 phenotype and did not require the auxiliary of other cells. In addition, puerarin suppressed the invasion and migration of NSCLC macrophages, restrained the expression of angiogenesis factors. Puerarin also inhibited the activation of mitogen-activated extracellular signal-regulated kinase (MEK)/extracellular signal-regulated kinase (ERK) 1/2 pathway through inhibition of ERK nucleus translocation. Finally, IL-4 induced M2 macrophage polarization and metastasis were partially offset by puerarin through inactivating the MEK/ERK 1/2 pathway. Taken together, this study validated that puerarin is able to skew macrophage populations back to M1 subsets to stimulate antitumor effects and suggests puerarin is a negative metastatic regulator of NSCLC.

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1	Kim R, Emi M, Tanabe K and Arihiro K: Tumor-driven evolution of immunosuppressive networks during malignant progression. Cancer Res. 66:5527–5536. 2006. View Article : Google Scholar : PubMed/NCBI
2	Harpole DH Jr: Prognostic modeling in early stage lung cancer: An evolving process from histopathology to genomics. Thorac Surg Clin. 17:167–173. viii2007. View Article : Google Scholar : PubMed/NCBI
3	Wood SL, Pernemalm M, Crosbie PA and Whetton AD: The role of the tumor-microenvironment in lung cancer-metastasis and its relationship to potential therapeutic targets. Cancer Treat Rev. 40:558–566. 2014. View Article : Google Scholar
4	Rhee I: Diverse macrophages polarization in tumor microenvironment. Arch Pharm Res. 39:1588–1596. 2016. View Article : Google Scholar : PubMed/NCBI
5	Shigeoka M, Urakawa N, Nishio M, Takase N, Utsunomiya S, Akiyama H, Kakeji Y, Komori T, Koma Y and Yokozaki H: Cyr61 promotes CD204 expression and the migration of macrophages via MEK/ERK pathway in esophageal squamous cell carcinoma. Cancer Med. 4:437–446. 2015. View Article : Google Scholar : PubMed/NCBI
6	Liu B, Wang X, Chen TZ, Li GL, Tan CC, Chen Y and Duan SQ: Polarization of M1 tumor associated macrophage promoted by the activation of TLR3 signal pathway. Asian Pac J Trop Med. 9:484–488. 2016. View Article : Google Scholar : PubMed/NCBI
7	Almatroodi SA, McDonald CF, Darby IA and Pouniotis DS: Characterization of M1/M2 tumour-associated macrophages (TAMs) and Th1/Th2 cytokine profiles in patients with NSCLC. Cancer Microenviron. 9:1–11. 2016. View Article : Google Scholar :
8	Ferrante CJ, Pinhal-Enfield G, Elson G, Cronstein BN, Hasko G, Outram S and Leibovich SJ: The adenosine-dependent angiogenic switch of macrophages to an M2-like phenotype is independent of interleukin-4 receptor alpha (IL-4Rα) signaling. Inflammation. 36:921–931. 2013. View Article : Google Scholar : PubMed/NCBI
9	Moore SM, Holt VV, Malpass LR, Hines IN and Wheeler MD: Fatty acid-binding protein 5 limits the anti-inflammatory response in murine macrophages. Mol Immunol. 67:265–275. 2015. View Article : Google Scholar : PubMed/NCBI
10	Choi JW, Kwon MJ, Kim IH, Kim YM, Lee MK and Nam TJ: Pyropia yezoensis glycoprotein promotes the M1 to M2 macrophage phenotypic switch via the STAT3 and STAT6 transcription factors. Int J Mol Med. 38:666–674. 2016.PubMed/NCBI
11	Tran TH, Rastogi R, Shelke J and Amiji MM: Modulation of macrophage functional polarity towards anti-inflammatory phenotype with plasmid DNA delivery in CD44 targeting hyaluronic acid nanoparticles. Sci Rep. 5:166322015. View Article : Google Scholar : PubMed/NCBI
12	Ohri CM, Shikotra A, Green RH, Waller DA and Bradding P: Macrophages within NSCLC tumour islets are predominantly of a cytotoxic M1 phenotype associated with extended survival. Eur Respir J. 33:118–126. 2009. View Article : Google Scholar : PubMed/NCBI
13	da Silva MD, Bobinski F, Sato KL, Kolker SJ, Sluka KA and Santos AR: IL-10 cytokine released from M2 macrophages is crucial for analgesic and anti-inflammatory effects of acupuncture in a model of inflammatory muscle pain. Mol Neurobiol. 51:19–31. 2015. View Article : Google Scholar
14	Zhang B, Yao G, Zhang Y and Gao J, Yang B, Rao Z and Gao J: M2-polarized tumor-associated macrophages are associated with poor prognoses resulting from accelerated lymphangiogenesis in lung adenocarcinoma. Clinics (Sao Paulo). 66:1879–1886. 2011. View Article : Google Scholar
15	Chen PC, Cheng HC, Wang J, Wang SW, Tai HC, Lin CW and Tang CH: Prostate cancer-derived CCN3 induces M2 macrophage infiltration and contributes to angiogenesis in prostate cancer microenvironment. Oncotarget. 5:1595–1608. 2014. View Article : Google Scholar : PubMed/NCBI
16	Zhang S, Chen S, Shen Y, Yang D, Liu X, Sun-Chi AC and Xu H: Puerarin induces angiogenesis in myocardium of rat with myocardial infarction. Biol Pharm Bull. 29:945–950. 2006. View Article : Google Scholar : PubMed/NCBI
17	Zhou YX, Zhang H and Peng C: Puerarin: A review of pharmacological effects. Phytother Res. 28:961–975. 2014. View Article : Google Scholar
18	Chen T, Chen H, Wang Y and Zhang J: In vitro and in vivo antitumour activities of puerarin 6″-O-xyloside on human lung carcinoma A549 cell line via the induction of the mitochondria-mediated apoptosis pathway. Pharm Biol. 5:1–7. 2016.
19	Liu C, Li Y, Yu J, Feng L, Hou S, Liu Y, Guo M, Xie Y, Meng J, Zhang H, et al: Targeting the shift from M1 to M2 macrophages in experimental autoimmune encephalomyelitis mice treated with fasudil. PLoS One. 8:e548412013. View Article : Google Scholar : PubMed/NCBI
20	Becker M, Müller CB, De Bastiani MA and Klamt F: The prognostic impact of tumor-associated macrophages and intra-tumoral apoptosis in non-small cell lung cancer. Histol Histopathol. 29:21–31. 2014.
21	Deepak P, Kumar S and Acharya A: Interleukin-13 neutralization modulates interleukin-13 induced suppression of reactive oxygen species production in peritoneal macrophages in a murine T-cell lymphoma. Cell Immunol. 251:72–77. 2008. View Article : Google Scholar : PubMed/NCBI
22	Zhang J, Cao J, Ma S, Dong R, Meng W, Ying M, Weng Q, Chen Z, Ma J, Fang Q, et al: Tumor hypoxia enhances non-small cell lung cancer metastasis by selectively promoting macrophage M2 polarization through the activation of ERK signaling. Oncotarget. 5:9664–9677. 2014. View Article : Google Scholar : PubMed/NCBI
23	Park SJ, Lee KP, Kang S, Lee J, Sato K, Chung HY, Okajima F and Im DS: Sphingosine 1-phosphate induced anti-atherogenic and atheroprotective M2 macrophage polarization through IL-4. Cell Signal. 26:2249–2258. 2014. View Article : Google Scholar : PubMed/NCBI
24	Ferreira R, Lively S and Schlichter LC: IL-4 type 1 receptor signaling up-regulates KCNN4 expression, and increases the KCa3.1 current and its contribution to migration of alternative-activated microglia. Front Cell Neurosci. 8:1832014. View Article : Google Scholar : PubMed/NCBI
25	Ji P, Diederichs S, Wang W, Böing S, Metzger R, Schneider PM, Tidow N, Brandt B, Buerger H, Bulk E, et al: MALAT-1, a novel noncoding RNA, and thymosin beta4 predict metastasis and survival in early-stage non-small cell lung cancer. Oncogene. 22:8031–8041. 2003. View Article : Google Scholar : PubMed/NCBI
26	Shi L, Zhang B, Sun X, Lu S, Liu Z, Liu Y, Li H, Wang L, Wang X and Zhao C: MiR-204 inhibits human NSCLC metastasis through suppression of NUAK1. Br J Cancer. 111:2316–2327. 2014. View Article : Google Scholar : PubMed/NCBI
27	Wang J, Yang ZR, Guo XF, Song J, Zhang JX, Wang J and Dong WG: Synergistic effects of puerarin combined with 5-fluorouracil on esophageal cancer. Mol Med Rep. 10:2535–2541. 2014.PubMed/NCBI
28	Guo XF, Yang ZR, Wang J, Lei XF, Lv XG and Dong WG: Synergistic antitumor effect of puerarin combined with 5-fluorouracil on gastric carcinoma. Mol Med Rep. 11:2562–2568. 2015.
29	Zeng YP, Yang ZR, Guo XF, Jun W and Dong WG: Synergistic effect of puerarin and 5-fluorouracil on hepatocellular carcinoma. Oncol Lett. 8:2436–2442. 2014.PubMed/NCBI
30	Wang Y, Ma Y, Zheng Y, Song J, Yang X, Bi C, Zhang D and Zhang Q: In vitro and in vivo anticancer activity of a novel puerarin nanosuspension against colon cancer, with high efficacy and low toxicity. Int J Pharm. 441:728–735. 2013. View Article : Google Scholar
31	Ruffell B, Affara NI and Coussens LM: Differential macrophage programming in the tumor microenvironment. Trends Immunol. 33:119–126. 2012. View Article : Google Scholar : PubMed/NCBI
32	Li D, Duan M, Feng Y, Geng L, Li X and Zhang W: MiR-146a modulates macrophage polarization in systemic juvenile idiopathic arthritis by targeting INHBA. Mol Immunol. 77:205–212. 2016. View Article : Google Scholar : PubMed/NCBI
33	Yang L, Wang F, Wang L, Huang L, Wang J, Zhang B and Zhang Y: CD163⁺ tumor-associated macrophage is a prognostic biomarker and is associated with therapeutic effect on malignant pleural effusion of lung cancer patients. Oncotarget. 6:10592–10603. 2015. View Article : Google Scholar : PubMed/NCBI
34	Deng X, Zhang P, Liang T, Deng S, Chen X and Zhu L: Ovarian cancer stem cells induce the M2 polarization of macrophages through the PPARγ and NF-κB pathways. Int J Mol Med. 36:449–454. 2015.PubMed/NCBI
35	Dinapoli MR, Calderon CL and Lopez DM: The altered tumoricidal capacity of macrophages isolated from tumor-bearing mice is related to reduce expression of the inducible nitric oxide synthase gene. J Exp Med. 183:1323–1329. 1996. View Article : Google Scholar : PubMed/NCBI
36	de Gaetano M, Crean D, Barry M and Belton O: M1- and M2-type macrophage responses are predictive of adverse outcomes in human atherosclerosis. Front Immunol. 7:2752016. View Article : Google Scholar : PubMed/NCBI
37	Belardelli F and Ferrantini M: Cytokines as a link between innate and adaptive antitumor immunity. Trends Immunol. 23:201–208. 2002. View Article : Google Scholar : PubMed/NCBI
38	Seo N and Tokura Y: Downregulation of innate and acquired antitumor immunity by bystander gammadelta and alphabeta T lymphocytes with Th2 or Tr1 cytokine profiles. J Interferon Cytokine Res. 19:555–561. 1999. View Article : Google Scholar : PubMed/NCBI
39	Iwamoto S, Iwai S, Tsujiyama K, Kurahashi C, Takeshita K, Naoe M, Masunaga A, Ogawa Y, Oguchi K and Miyazaki A: TNF-alpha drives human CD14⁺ monocytes to differentiate into CD70⁺ dendritic cells evoking Th1 and Th17 responses. J Immunol. 179:1449–1457. 2007. View Article : Google Scholar : PubMed/NCBI
40	Zhang F, Wang H, Wang X, Jiang G, Liu H, Zhang G, Wang H, Fang R, Bu X, Cai S, et al: TGF-β induces M2-like macrophage polarization via SNAIL-mediated suppression of a pro-inflammatory phenotype. Oncotarget. 13:105612016.
41	Deng B, Wehling-Henricks M, Villalta SA, Wang Y and Tidball JG: IL-10 triggers changes in macrophage phenotype that promote muscle growth and regeneration. J Immunol. 189:3669–3680. 2012. View Article : Google Scholar : PubMed/NCBI
42	Han J, Yu CQ and Shen W: Inhibitory effects of puerarin on invasion and metastasis of oophoroma cells HO-8910. Zhongguo Zhong Xi Yi Jie He Za Zhi. 29:632–635. 2009.In Chinese. PubMed/NCBI
43	Wang D, Liu Y, Han J, Zai D, Ji M, Cheng W, Xu L, Yang L, He M, Ni J, et al: Puerarin suppresses invasion and vascularization of endometriosis tissue stimulated by 17β-estradiol. PLoS One. 6:e250112011. View Article : Google Scholar
44	Lee SH, Jaganath IB, Manikam R and Sekaran SD: Inhibition of Raf-MEK-ERK and hypoxia pathways by Phyllanthus prevents metastasis in human lung (A549) cancer cell line. BMC Complement Altern Med. 13:2712013. View Article : Google Scholar : PubMed/NCBI