[논문]Ethyl Acetate Fraction of Adenophora triphylla var. japonica Inhibits Migration of Lewis Lung Carcinoma Cells by Suppressing Macrophage Polarization toward an M2 Phenotype

Park, Shin-Hyung

doi:10.3831/kpi.2019.22.034

Ethyl Acetate Fraction of Adenophora triphylla var. japonica Inhibits Migration of Lewis Lung Carcinoma Cells by Suppressing Macrophage Polarization toward an M2 Phenotype 원문보기

Journal of pharmacopuncture, v.22 no.4, 2019년, pp.253 - 259

Park, Shin-Hyung (Departments of Pathology, College of Korean Medicine, Dong-eui University)

Abstract ▼ AI-Helper

Objectives: It is reported that tumor-associated macrophages (TAMs) contribute to cancer progression by promoting tumor growth and metastasis. The purpose of this study is to investigate the effect of different fractions of Adenophora triphylla var. japonica (AT) on the polarization of macrophages into the M2 phenotype, a major phenotype of TAMs. Methods: We isolated hexane, ethyl acetate, and butanol fractions from crude ethanol extract of AT. The cytotoxicity of AT in RAW264.7 cells was examined by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. RAW264.7 cells were polarized into the M2 phenotype by treatment with interleukin (IL)-4 and IL-13. The expression of M2 macrophage marker genes was detected by reverse transcription polymerase chain reaction (RT-PCR). The phosphorylation level of signal transducer and activator of transcription 6 (STAT6) was investigated by western blot analysis. The migration of Lewis lung carcinoma (LLC) cells was examined by transwell migration assay using conditioned media (CM) collected from RAW264.7 cells as a chemoattractant. Results: Among various fractions of AT, the ethyl acetate fraction of AT (EAT) showed the most significant suppressive effect on the mRNA expression of M2 macrophage markers, including arginase-1, interleukin (IL)-10 and mannose receptor C type 1 (MRC-1), up-regulated by treatment of IL-4 and IL-13. In addition, EAT suppressed the phosphorylation of STAT6, a critical regulator of IL-4 and IL-13-induced M2 macrophage polarization. Finally, the increased migration of Lewis lung carcinoma (LLC) cells by CM from M2-polarized RAW264.7 cells was reduced by CM from RAW264.7 cells co-treated with EAT and M2 polarization inducers. Conclusion: We demonstrated that EAT attenuated cancer cell migration through suppression of macrophage polarization toward the M2 phenotype. Additional preclinical or clinical researches are needed to evaluate its regulatory effects on macrophage polarization and anti-cancer activities.

주제어

표/그림 (5)

$Figure 1 Effects of the AT subfractions on the cell viability in RAW264.7 cells$ 그림 Figure 1 Effects of the AT subfractions on the cell viability in RAW264.7 cells
$Figure 2 Effects of the AT subfractions on the expression of M2 marker genes in RAW264.7 cells$ 그림 Figure 2 Effects of the AT subfractions on the expression of M2 marker genes in RAW264.7 cells
그림 Figure 3 Effects of EAT on the expression of M2 marker genes in RAW264.7 cells
그림 Figure 4 Effects of EAT on the phosphorylation level of STAT6 in RAW264.7 cells
그림 Figure 5 Effects of EAT on the migration of Lewis lung carcinoma cells

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가설 설정

Although the previous studies reporting the anti-cancer effects of AT mainly focused on the cancer cell regulation, the influence of AT on the TME regulation has not been explored yet. Given that the extracts and constituents of AT commonly exhibited anti-inflammatory activities by regulating macrophages, we hypothesized that the extracts of AT would influence macrophage polarization. Therefore, the current study investigated the effects of different fractions of AT on macrophage polarization into the M2 phenotype.

제안 방법

Thus, we used 50 µg/mL and 200 µg/mL as the maximum concentration of HAT/BAT and EAT, respectively, for further experiments.

대상 데이터

Primary antibodies against phospho- signal transducer and activator of transcription 6 (STAT6), total-STAT6, and actin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-rabbit secondary antibody and anti-mouse secondary antibody were purchased from Enzo Life Sciences (Farmingdale, USA) and Bethyl Laboratories (Montgomery, TX), respectively.

데이터처리

Significance was determined by the Student's t-test (*** P < 0.001 vs. respective control).
Statistical significance was estimated by Student’s t-test.

이론/모형

7 mouse macrophage cells were treated with the three subfractions of AT for 24 h. The cell viability was evaluated by MTT assay. Data are expressed as the mean ± S.

성능/효과

In conclusion, our study demonstrated that EAT suppressed the macrophage polarization towards the M2 phenotype, leading to the attenuation of cancer cell migration. We provide a valuable information for finding new anticancer materials from traditional herbal medicines based on their TAM-modulatory effects.
The results showed that EAT treatment down-regulated the mRNA expression of arginase-1, IL-10, and MRC-1 in a concentration-dependent manner even though IL-10 level was slightly recovered at 200 µg/mL (Figure 3A and 3B).

참고문헌 (25)

1 Hanahan D Coussens LM Accessories to the crime: functions of cells recruited to the tumor microenvironment Cancer Cell 2012 21 3 309 22 10.1016/j.ccr.2012.02.022 22439926

상세보기
2 Pollard JW Tumour-educated macrophages promote tumour progression and metastasis Nat Rev Cancer 2004 4 1 71 8 10.1038/nrc1256 14708027

상세보기
3 Mei J Xiao Z Guo C Pu Q Ma L Liu C Prognostic impact of tumor-associated macrophage infiltration in non-small cell lung cancer: A systemic review and meta-analysis Oncotarget 2016 7 23 34217 28 10.18632/oncotarget.9079 27144518

상세보기
4 Brown JM Recht L Strober S The Promise of Targeting Macrophages in Cancer Therapy Clin Cancer Res 2017 23 13 3241 50 10.1158/1078-0432.CCR-16-3122 28341752

상세보기
5 Jang IM Treatise on Asian herbal medicines Seoul Haksul-pyunsu-kwan in Research Institute of Natural Products of Seoul National University 2003 Korean
6 Geng G Sun FY Efficacy of Yiqi Yangyin in the treatment of advanced non-small cell lung cancer Nei Mongol Journal of Traditional Chinese Medicine 1991 4 4 5
7 Ma CY Tang YJ Application of Yiqi Yangyin in lung cancer Journal of Shandong college of traditional chinese medicine 1996 20 3 178 9
8 Zhang T Wang B Clinical study on integrated Chinese and western medicine combined with radiotherapy in treatment of patients with advanced lung cancer Journal of Clinical and Experimental Medicine 2013 12 2 108 10
9 Liang F Zhang Y Li CH Cao HT Effect of Yiqi Yangyin Jiedu Formula on T-Cell Subsets and Shifting of Th1/Th2 Balance in NSCLC Patients Journal of Practical Traditional Chinese Internal Medicine 2011 25 5 8 10
10 Lee DR Lee YS Choi BK Lee HJ Park SB Kim TM Roots extracts of Adenophora triphylla var. japonica improve obesity in 3T3-L1 adipocytes and high-fat diet-induced obese mice Asian Pac J Trop Med 2015 8 11 898 906 10.1016/j.apjtm.2015.10.011 26614988

상세보기
11 Hu JR Jung CJ Ku SM Jung DH Ku SK Choi JS Anti-tussive, expectorant, and anti-inflammatory effects of Adenophorae Radix powder in ICR mice J Ethnopharmacol 2019 239 111915 10.1016/j.jep.2019.111915 31039428

상세보기
12 Jin SE Son YK Min BS Jung HA Choi JS Anti-inflammatory and antioxidant activities of constituents isolated from Pueraria lobata roots Arch Pharm Res 2012 35 5 823 37 10.1007/s12272-012-0508-x 22644850

원문보기 상세보기
13 Kang SC Lim SY Song YJ Lupeol is one of active components in the extract of Chrysanthemum indicum Linne that inhibits LMP1-induced NF-kappaB activation PLoS One 2013 8 11 e82688 10.1371/journal.pone.0082688 24303085

상세보기
14 Yao X Li G Bai Q Xu H Lu C Taraxerol inhibits LPS-induced inflammatory responses through suppression of TAK1 and Akt activation Int Immunopharmacol 2013 15 2 316 24 10.1016/j.intimp.2012.12.032 23333629

상세보기
15 Srivastava P Jyotshna Gupta N Maurya AK Shanker K New anti-inflammatory triterpene from the root of Ricinus communis Nat Prod Res 2014 28 5 306 11 10.1080/14786419.2013.861834 24279342

상세보기
16 Yoon YP Lee HJ Lee DU Lee SK Hong JH Lee CJ Effects of Lupenone, Lupeol, and Taraxerol Derived from Adenophora triphylla on the Gene Expression and Production of Airway MUC5AC Mucin Tuberc Respir Dis (Seoul) 2015 78 3 210 7 10.4046/trd.2015.78.3.210 26175774

원문보기 상세보기
17 Ovesna Z Vachalkova A Horvathova K Taraxasterol and beta-sitosterol: new naturally compounds with chemoprotective/chemopreventive effects Neoplasma 2004 51 6 407 14 15640948

상세보기
18 Chun J Kang M Kim YS A triterpenoid saponin from Adenophora triphylla var. japonica suppresses the growth of human gastric cancer cells via regulation of apoptosis and autophagy Tumour Biol 2014 35 12 12021 30 10.1007/s13277-014-2501-0 25146681

상세보기
19 Kang M Ha IJ Chun J Kang SS Kim YS Separation of two cytotoxic saponins from the roots of Adenophora triphylla var. japonica by high-speed counter-current chromatography Phytochem Anal 2013 24 2 148 54 10.1002/pca.2394 22930642

상세보기
20 Bi Y Chen J Hu F Liu J Li M Zhao L M2 Macrophages as a Potential Target for Antiatherosclerosis Treatment Neural Plast 2019 2019 6724903 10.1155/2019/6724903 30923552

상세보기
21 Solinas G Germano G Mantovani A Allavena P Tumor-associated macrophages (TAM) as major players of the cancer-related inflammation J Leukoc Biol 2009 86 5 1065 73 10.1189/jlb.0609385 19741157

상세보기
22 Singh N Baby D Rajguru JP Patil PB Thakkannavar SS Pujari VB Inflammation and cancer Ann Afr Med 2019 18 3 121 6 10.4103/aam.aam_56_18 31417011

상세보기
23 Finn OJ Immuno-oncology: understanding the function and dysfunction of the immune system in cancer Ann Oncol 2012 23 Suppl 8 viii6 9 10.1093/annonc/mds256 22918931

상세보기
24 Choi HJ Choi HJ Chung TW Ha KT Luteolin inhibits recruitment of monocytes and migration of Lewis lung carcinoma cells by suppressing chemokine (C-C motif) ligand 2 expression in tumor-associated macrophage Biochem Biophys Res Commun 2016 470 1 101 6 10.1016/j.bbrc.2016.01.002 26766793

상세보기
25 Oncotarget 2015 11 3 6 34 36441 55 10.18632/oncotarget.5541 Metformin prevents cancer metastasis by inhibiting M2-like polarization of tumor associated macrophages Ding L 1 Liang G 1 Yao Z 1 Zhang J 1 Liu R 1 Chen H 1 Zhou Y 1 Wu H 1 Yang B 1 He Q 1 26497364

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