Bleomycin inhibits proliferation and induces apoptosis in TPC‑1 cells through reversing M2‑macrophages polarization

Liu,Hongwei; Dong,Huilei; Jiang,Li; Li,Zhendong; Ma,Xiande

doi:10.3892/ol.2018.9103

Bleomycin inhibits proliferation and induces apoptosis in TPC‑1 cells through reversing M2‑macrophages polarization

Authors:
- Hongwei Liu
- Huilei Dong
- Li Jiang
- Zhendong Li
- Xiande Ma
View Affiliations

Affiliations: Department of Head and Neck Surgery, Cancer Hospital of China Medical University, Shenyang, Liaoning 110042, P.R. China, College of Integrated Traditional Chinese and Western Medicine, Liaoning University of Traditional Chinese Medicine, Shenyang, Liaoning 110847, P.R. China
Published online on: July 6, 2018 https://doi.org/10.3892/ol.2018.9103
Pages: 3858-3866
Copyright: © Liu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Papillary thyroid carcinoma (PTC) is one of the most common types of thyroid malignancy. Previous studies have demonstrated that the density of tumor‑associated macrophages (TAMs) within the tumor microenvironment affects the progression of PTC due to the imbalance in M1/M2 macrophage subtypes. M2 macrophages induce anti‑inflammatory effects and promote tumor progression, whereas M1 macrophages destroy tumor cells. Therefore, reversing TAM polarization to M1 may be a novel strategy for the treatment of cancer. Although bleomycin (BLM) is a commonly used anti‑cancer drug, which regulates the secretion of relevant cytokines, high dose and long‑term treatment with BLM may lead to pulmonary fibrosis. In the present study, flow cytometry data revealed that low dose treatment with BLM (5 or 10 mU/ml) facilitated the expression of the M1 phenotype markers cluster of differentiation (CD)80 and C‑C chemokine receptor 7, and concurrently suppressed the M2 marker CD206 on M2‑macrophages. Reverse transcription‑quantitative polymerase chain reaction data revealed that the expression levels of tumor necrosis factor‑α and interleukin‑1β markedly increased, whereas the expression of IL‑10 decreased in M2 macrophages treated with BLM. A fluorescein isothiocyanate‑dextran uptake experiment revealed that BLM increased the phagocytic capacity of M2, however not M1 or M0 macrophages. In addition, to verify the effect of BLM‑treated M2 macrophages on thyroid carcinoma cells, a co‑culture system of macrophages and the human PTC cell line TPC‑1, was established. BLM‑treated M2 macrophages increased the number of cells in early and late apoptosis and inhibited the migration, proliferation and invasion of TPC‑1 cells. These results suggest that a low dose and indirect effect of BLM may induce suppressive effects on PTC by selectively reversing M2 macrophage polarization to M1, which may provide a novel strategy for cancer treatment.

View Figures

View References

1	Cancer Genome Atlas Network: Integrated genomic characterization of papillary thyroid carcinoma. Cell. 159:676–690. 2014. View Article : Google Scholar : PubMed/NCBI
2	Walker S: Immunotherapy and the treatment of non-small cell lung cancer. J Adv Pract Oncol. 7:304–306. 2016.PubMed/NCBI
3	Pusztaszeri MP, Faquin WC and Sadow PM: Tumor-associated inflammatory cells in thyroid carcinomas. Surg Pathol Clin. 7:501–514. 2014. View Article : Google Scholar : PubMed/NCBI
4	Qing W, Fang WY, Ye L, Shen LY, Zhang XF, Fei XC, Chen X, Wang WQ, Li XY, Xiao JC and Ning G: Density of tumor-associated macrophages correlate with lymph node metastasis in papillary thyroid carcinoma. Thyroid. 22:905–910. 2012. View Article : Google Scholar : PubMed/NCBI
5	Quatromoni JG and Eruslanov E: Tumor-associated macrophages: Function, phenotype and link to prognosis in human Lung Cancer. Am J Transl Res. 4:376–389. 2012.PubMed/NCBI
6	Mantovani A, Sozzani S, Locati M, Allavena P and Sica A: Macrophage polarization: Tumor-associated macrophages as a paradigm for polarized M2 mononuclear phagocytes. Trends Immunol. 23:549–555. 2002. View Article : Google Scholar : PubMed/NCBI
7	Allavena P, Sica A, Solinas G, Porta C and Mantovani A: The inflammatory micro-environment in tumor progression: The role of tumor-associated macrophages. Crit Rev Oncol Hematol. 66:1–9. 2008. View Article : Google Scholar : PubMed/NCBI
8	Can NY, Ayturk S, Celik M, Sezer YA, Ozyilmaz F, Tastekin E, Sut N, Ustun F, Bulbul BY, Puyan FO and Guldiken S: Histological perspective on the effects of tumor-associated macrophages in the tumor microenvironment surrounding papillary thyroid carcinoma. Pol J Pathol. 67:332–344. 2016. View Article : Google Scholar : PubMed/NCBI
9	Fang W, Ye L, Shen L, Cai J, Huang F, Wei Q, Fei X, Chen X, Guan H, Wang W, et al: Tumor-associated macrophages promote the metastatic potential of thyroid papillary cancer by releasing CXCL8. Carcinogenesis. 35:1780–1787. 2014. View Article : Google Scholar : PubMed/NCBI
10	Chang WC, Chen JY, Lee CH and Yang AH: Expression of decoy receptor 3 in diffuse sclerosing variant of papillary thyroid carcinoma: Correlation with M2 macrophage differentiation and lymphatic invasion. Thyroid. 23:720–726. 2013. View Article : Google Scholar : PubMed/NCBI
11	Lopez-Gonzalez JS, Avila-Moreno F, Prado-Garcia H, Aguilar-Cazares D, Mandoki JJ and Meneses-Flores M: Lung carcinomas decrease the number of monocytes/macrophages (CD14+ cells) that produce TNF-alpha. Clin Immunol. 122:323–329. 2007. 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	Redente EF, Dwyer-Nield LD, Merrick DT, Raina K, Agarwal R, Pao W, Rice PL, Shroyer KR and Malkinson AM: Tumor progression stage and anatomical site regulate tumor-associated macrophage and bone marrow-derived monocyte polarization. Am J Pathol. 176:2972–2985. 2010. View Article : Google Scholar : PubMed/NCBI
14	Zea AH, Rodriguez PC, Atkins MB, Hernandez C, Signoretti S, Zabaleta J, McDermott D, Quiceno D, Youmans A, O'Neill A, et al: Arginase-producing myeloid suppressor cells in renal cell carcinoma patients: A mechanism of tumor evasion. Cancer Res. 65:3044–3048. 2005. View Article : Google Scholar : PubMed/NCBI
15	Chang CI, Liao JC and Kuo L: Macrophage arginase promotes tumor cell growth and suppresses nitric oxide-mediated tumor cytotoxicity. Cancer Res. 61:1100–1106. 2001.PubMed/NCBI
16	Biswas SK, Sica A and Lewis CE: Plasticity of macrophage function during tumor progression: Regulation by distinct molecular mechanisms. J Immunol. 180:2011–2057. 2008. View Article : Google Scholar : PubMed/NCBI
17	Qian BZ and Pollard JW: Macrophage diversity enhances tumor progression and metastasis. Cell. 141:39–51. 2010. View Article : Google Scholar : PubMed/NCBI
18	Mehrpour M, Esclatine A, Beau I and Codogno P: Overview of macroautophagy regulation in mammalian cells. Cell Res. 20:748–762. 2010. View Article : Google Scholar : PubMed/NCBI
19	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
20	Batschinski K, Dervisis N, Kitchell B, Newman R and Erfourth T: Combination of bleomycin and cytosine arabinoside chemotherapy for relapsed canine lymphoma. J Am Anim Hosp Assoc. 54:150–155. 2018. View Article : Google Scholar : PubMed/NCBI
21	Ng K, Duncan S, Shamash J and Alifrangis C: Dose intense chemotherapy in the management of poor prognosis and relapsed testicular cancer: Experiences and controversies. Expert Rev Anticancer Ther. 18:431–436. 2018. View Article : Google Scholar : PubMed/NCBI
22	Porwal PK, Dubey KP, Morey A, Singh H, Pooja S and Bose A: Bleomycin sclerotherapy in lymphangiomas of head and neck: prospective study of 8 cases. Indian J Otolaryngol Head Neck Surg. 70:145–148. 2018. View Article : Google Scholar : PubMed/NCBI
23	Racnik J, Svara T, Zadravec M, Gombac M, Cemazar M, Sersa G and Tozon N: Electrochemotherapy with bleomycin of different types of cutaneous tumours in a ferret (Mustela Putorius Furo). Radiol Oncol. 52:98–104. 2017. View Article : Google Scholar : PubMed/NCBI
24	Cai Y, Sun R, Wang R, Ren JG, Zhang W, Zhao YF and Zhao JH: The activation of Akt/mTOR pathway by bleomycin in Epithelial-to-mesenchymal transition of human submandibular gland cells: A treatment mechanism of bleomycin for mucoceles of the salivary glands. Biomed Pharmacother. 90:109–115. 2017. View Article : Google Scholar : PubMed/NCBI
25	Mouratis MA and Aidinis V: Modeling pulmonary fibrosis with bleomycin. Curr Opin Pulm Med. 17:355–361. 2011. View Article : Google Scholar : PubMed/NCBI
26	Weng D, Chen JX, Li HH, Liu F, Zhou LD, Liu HP, Zheng RJ, Jiang Y, Liu ZH and Ge B: 2-aminopurine suppresses the TGF-β1-induced epithelial-mesenchymal transition and attenuates bleomycin-induced pulmonary fibrosis. Cell Death Discov. 4:172018. View Article : Google Scholar : PubMed/NCBI
27	Jung WJ, Lee SY, Choi SI, Kim BK, Lee EJ, In KH and Lee MG: Toll-like receptor expression in pulmonary sensory neurons in the bleomycin-induced fibrosis model. PLoS One. 13:e01931172018. View Article : Google Scholar : PubMed/NCBI
28	Bolzan AD and Bianchi MS: DNA and chromosome damage induced by bleomycin in mammalian cells: An update. Mutat Res. 775:51–62. 2018. View Article : Google Scholar : PubMed/NCBI
29	Massonneau J, Ouellet C, Lucien F, Dubois CM, Tyler J and Boissonneault G: Suboptimal extracellular pH values alter DNA damage response to induced double-strand breaks. FEBS Open Bio. 8:416–425. 2018. View Article : Google Scholar : PubMed/NCBI
30	Huang W, Wang G, Phelps DS, Al-Mondhiry H and Floros J: Combined SP-A-bleomycin effect on cytokines by THP-1 cells: Impact of surfactant lipids on this effect. Am J Physiol Lung Cell Mol Physiol. 283:L94–L102. 2002. View Article : Google Scholar : PubMed/NCBI
31	Razonable RR, Henault M and Paya CV: Stimulation of toll-like receptor 2 with bleomycin results in cellular activation and secretion of pro-inflammatory cytokines and chemokines. Toxicol Appl Pharmacol. 210:181–189. 2006. View Article : Google Scholar : PubMed/NCBI
32	Hoshino T, Okamoto M, Sakazaki Y, Kato S, Young HA and Aizawa H: Role of proinflammatory cytokines IL-18 and IL-1beta in bleomycin-induced lung injury in humans and mice. Am J Respir Cell Mol Biol. 41:661–670. 2009. View Article : Google Scholar : PubMed/NCBI
33	Genin M, Clement F, Fattaccioli A, Raes M and Michiels C: M1 and M2 macrophages derived from THP-1 cells differentially modulate the response of cancer cells to etoposide. BMC Cancer. 15:5772015. View Article : Google Scholar : PubMed/NCBI
34	Steinbach F and Thiele B: Phenotypic investigation of mononuclear phagocytes by flow cytometry. J Immunol Methods. 174:109–122. 1994. View Article : Google Scholar : PubMed/NCBI
35	Murdoch C, Muthana M, Coffelt SB and Lewis CE: The role of myeloid cells in the promotion of tumour angiogenesis. Nat Rev Cancer. 8:618–631. 2008. View Article : Google Scholar : PubMed/NCBI
36	Yuan A, Hsiao YJ, Chen HY, Chen HW, Ho CC, Chen YY, Liu YC, Hong TH, Yu SL, Chen JJ and Yang PC: Opposite effects of M1 and M2 macrophage subtypes on lung cancer progression. Sci Rep. 5:142732015. View Article : Google Scholar : PubMed/NCBI
37	Reeves AR, Spiller KL, Freytes DO, Vunjak-Novakovic G and Kaplan DL: Controlled release of cytokines using silk-biomaterials for macrophage polarization. Biomaterials. 73:272–283. 2015. View Article : Google Scholar : PubMed/NCBI
38	Pang L, Han S, Jiao Y, Jiang S, He X and Li P: Bu Fei Decoction attenuates the tumor associated macrophage stimulated proliferation, migration, invasion and immunosuppression of non-small cell Lung Cancer, partially via IL-10 and PD-L1 regulation. Int J Oncol. 51:25–38. 2017. View Article : Google Scholar : PubMed/NCBI
39	Kang H, Zhang J, Wang B, Liu M, Zhao J, Yang M and Li Y: Puerarin inhibits M2 polarization and metastasis of tumor-associated macrophages from NSCLC xenograft model via inactivating MEK/ERK 1/2 pathway. Int J Oncol. 50:545–554. 2017. View Article : Google Scholar : PubMed/NCBI
40	Cho SW, Kim YA, Sun HJ, Kim YA, Oh BC, Yi KH, Park DJ and Park YJ: CXCL16 signaling mediated macrophage effects on tumor invasion of papillary thyroid carcinoma. Endocr Relat Cancer. 23:113–124. 2016. View Article : Google Scholar : PubMed/NCBI
41	Komohara Y, Fujiwara Y, Ohnishi K and Takeya M: Tumor-associated macrophages: Potential therapeutic targets for anti-cancer therapy. Adv Drug Deliv Rev. 99:180–185. 2016. View Article : Google Scholar : PubMed/NCBI
42	Gocheva V, Wang HW, Gadea BB, Shree T, Hunter KE, Garfall AL, Berman T and Joyce JA: IL-4 induces cathepsin protease activity in tumor-associated macrophages to promote cancer growth and invasion. Genes Dev. 24:241–255. 2010. View Article : Google Scholar : PubMed/NCBI
43	Wang R, Zhang J, Chen S, Lu M, Luo X, Yao S, Liu S, Qin Y and Chen H: Tumor-associated macrophages provide a suitable microenvironment for non-small Lung Cancer invasion and progression. Lung Cancer. 74:188–196. 2011. View Article : Google Scholar : PubMed/NCBI
44	Komohara Y, Jinushi M and Takeya M: Clinical significance of macrophage heterogeneity in human malignant tumors. Cancer Sci. 105:1–8. 2014. View Article : Google Scholar : PubMed/NCBI
45	Rolny C, Mazzone M, Tugues S, Laoui D, Johansson I, Coulon C, Squadrito ML, Segura I, Li X, Knevels E, et al: HRG inhibits tumor growth and metastasis by inducing macrophage polarization and vessel normalization through downregulation of PlGF. Cancer Cell. 19:31–44. 2011. View Article : Google Scholar : PubMed/NCBI
46	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 : PubMed/NCBI
47	Muraille E, Leo O and Moser M: TH1/TH2 paradigm extended: Macrophage polarization as an unappreciated pathogen-driven escape mechanism. Front Immunol. 5:6032014. View Article : Google Scholar : PubMed/NCBI
48	Kim S, Cho SW, Min HS, Kim KM, Yeom GJ, Kim EY, Lee KE, Yun YG, Park DJ and Park YJ: The expression of tumor-associated macrophages in papillary thyroid carcinoma. Endocrinol Metab (Seoul). 28:192–198. 2013. View Article : Google Scholar : PubMed/NCBI
49	Mellor AL and Munn DH: Creating immune privilege: Active local suppression that benefits friends, but protects foes. Nat Rev Immunol. 8:74–80. 2008. View Article : Google Scholar : PubMed/NCBI
50	Sica A, Schioppa T, Mantovani A and Allavena P: Tumour-associated macrophages are a distinct M2 polarised population promoting tumour progression: Potential targets of anti-cancer therapy. Eur J Cancer. 42:717–727. 2006. View Article : Google Scholar : PubMed/NCBI
51	Liu CP, Zhang X, Tan QL, Xu WX, Zhou CY, Luo M, Li X, Huang RY and Zeng X: NF-κB pathways are involved in M1 polarization of RAW 264.7 macrophage by polyporus polysaccharide in the tumor microenvironment. PLoS One. 12:e01883172017. View Article : Google Scholar : PubMed/NCBI
52	Gong H, Cao Y, Han G, Zhang Y, You Q, Wang Y and Pan Y: p53/microRNA-374b/AKT1 regulates colorectal cancer cell apoptosis in response to DNA damage. Int J Oncol. 50:1785–1791. 2017. View Article : Google Scholar : PubMed/NCBI
53	Yang L, Zhao W, Wei P, Zuo W and Zhu S: Tumor suppressor p53 induces miR-15a processing to inhibit neuronal apoptosis inhibitory protein (NAIP) in the apoptotic response DNA damage in breast cancer cell. Am J Transl Res. 9:683–691. 2017.PubMed/NCBI
54	Liu JF, Nie XC, Shao YC, Su WH and Ma HY: Bleomycin suppresses the proliferation and the mobility of human gastric cancer cells through the smad signaling pathway. Cell Physiol Biochem. 40:1401–1409. 2016. View Article : Google Scholar : PubMed/NCBI
55	Gao N, Shang B, Zhang X, Shen C, Xu R, Chen R and He Q: Potent antitumor actions of the new antibiotic boningmycin through induction of apoptosis and cellular senescence. Anticancer Drugs. 22:166–175. 2011. View Article : Google Scholar : PubMed/NCBI
56	Uchida R, Yokota S and Tomoda H: Structure elucidation of meroterpenoid habiterpenol, a novel abrogator of bleomycin-induced G2 arrest in Jurkat cells, produced by Phytohabitans suffuscus 3787_5. J Antibiot (Tokyo). 67:783–786. 2014. View Article : Google Scholar : PubMed/NCBI
57	Wang Q, Cui K, Espin-Garcia O, Cheng D, Qiu X, Chen Z, Moore M, Bristow RG, Xu W, Der S and Liu G: Resistance to bleomycin in cancer cell lines is characterized by prolonged doubling time, reduced DNA damage and evasion of G2/M arrest and apoptosis. PLoS One. 8:e823632013. View Article : Google Scholar : PubMed/NCBI
58	Lofdahl A, Rydell-Tormanen K, Larsson-Callerfelt AK, AUID- and Wenglen Oho C: Pulmonary fibrosis in vivo displays increased p21 expression reduced by 5-HT2B receptor antagonists in vitro-a potential pathway affecting proliferation. Sci Rep. 8:19272018. View Article : Google Scholar : PubMed/NCBI
59	Park HR, Lee EJ, Moon SC, Chung TW and Kim KJ: Inhibition of lung cancer growth by HangAmDan-B is mediated by macrophage activation to M1 subtype. Oncol Lett. 13:2330–2336. 2017. View Article : Google Scholar : PubMed/NCBI
60	Li Y, Poppoe F, Chen J, Yu L and Deng F: Macrophages polarized by expression of toxoGRA15II inhibit growth of hepatic carcinoma. Front Immunol. 8:1372017.PubMed/NCBI