Evaluation of antitumor activity of Artemisia capillaris extract against hepatocellular carcinoma through the inhibition of IL-6/STAT3 signaling axis

  • Authors:
    • Eungyeong Jang
    • So-Young Kim
    • Na-Rae Lee
    • Chae-Min Yi
    • Da-Rong Hong
    • Woo Seok Lee
    • Jong-Ho Kim
    • Kyung-Tae Lee
    • Bum-Joon Kim
    • Jang-Hoon Lee
    • Kyung-Soo Inn
  • View Affiliations

  • Published online on: November 29, 2016     https://doi.org/10.3892/or.2016.5283
  • Pages: 526-532
Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Abstract

Interleukin-6 (IL-6)/signal transducer and activator of transcription 3 (STAT3) signaling pathway plays critical roles in the development and progression of hepatocellular carcinoma (HCC). Artemisia capillaris (AC) has been widely used to treat various liver diseases including HCC as a herbal medicine. The effects of AC on IL-6/STAT3 signaling axis in HCC cells and subsequent anticancer activity of AC against HCC were analyzed using HCC cell lines and HBV W4P-LHB-expressing NIH3T3 cell line, which has been shown to gain tumorigenicity by activating IL-6/STAT3 signaling in our previous study. AC extract significantly suppressed the growth and colony formation of HCC cells. In addition, it inhibited the activation of STAT3 by IL-6 and subsequent synthesis of downstream molecules in HCC and W4P-NIH3T3 cells. Consequently, migration of cells was significantly suppressed by the AC extract. Collectively, the findings suggest that AC extract is capable of conferring various antitumor effects against HCC through the modulation of the IL-6/STAT3 pathway. The results provide a basis for the therapeutic use of AC in the treatment of HCC. Identification of the compound responsible for the effect may lead to the development of a novel anticancer agent against HCC.

Introduction

The pathogenesis of hepatocellular carcinoma (HCC) is closely linked with chronic inflammation and the level of interleukin-6 (IL-6). Serum IL-6 level is higher in HCC patients than in healthy adults (1), demonstrating that inflammation is a critical risk factor for the formation and progression of HCC. IL-6 rapidly activates the signal transducer and activator of transcription 3 (STAT3), which is a major mediator regulating signal transduction from IL-6 to the nucleus and inducing the transcription of proliferation associated genes (2). Phosphorylated STAT3 has been implicated in abnormal oncogenic processes, such as initiation, proliferation, angiogenesis, and progression in HCC (3). Owing to these carcinogenetic roles of the IL-6/STAT3 signaling pathway, which promotes the development of HCC, STAT3 is often considered an attractive target for liver cancer therapy. In addition, in our previous study, we found that male-specific W4P mutations in the preS1 region contribute to carcinogenesis and male predominance in HCC via activation of the IL-6/STAT3 signaling axis. Ectopic expression of W4P mutant HBV large antigen (LHB) was sufficient to induce transformation of NIH3T3 fibroblast cells, emphasizing the crucial role of the IL-6/STAT3 signaling pathway in HCC carcinogenesis (4).

Artemisia is one of the most important genera of the Asteraceae or Compositae family, the largest family of flowering plants with over 15,000 species (5). The genus Artemisia consists of more than 400 species, some of which exhibit medicinal activities for a variety of diseases such as malaria, rabies, tonsillitis, asthma, gonorrhea, cough, syphilis, and leprosy (6). Amongst these species, Artemisia capillaris Thunberg (AC) has long been used in Asian countries as a therapeutic drug (7,8). According to previous studies on its medicinal properties, AC exhibits various pharmacological effects, such as antimicrobial (8), anti-obesitic (9,10), and antitumor (11) effects. In particular, there have been many studies on the therapeutic and pharmacological effects of AC against liver diseases, including hepatoprotective effects in obese mice (12) or ethanol-administered mice (13), antiviral effects by suppressing the replication and secretion of the hepatitis B virus (1416), and anti-fibrotic effects in animal models (7,17,18).

HCC, the most common primary liver cancer, demands more effective remedies because of a low 5-year survival rate, about 10% (19), and poor reactions refractory to available treatments (20). Increasing evidence indicates the efficacy of AC in suppressing the proliferation of human hepatoma cells (2123). In a recent study, capillarisin derived from AC was found to have anticancer effects by suppressing STAT3 phosphorylation in human multiple myeloma cells (24). However, there have been no previous studies evaluating the anticancer effects of AC via inhibition of the STAT3 signaling pathway in vitro models of HCC.

Therefore, in this study, we examined the possibility of AC exerting its antitumor activity against HCC by modulating IL-6-induced STAT3 activation.

Materials and methods

Preparation of AC extract (ACE)

AC was purchased from Kyung Hee Herb Pharm (Wonju, Korea), a licensed herb company. AC was pulverized and extracted with 70% EtOH under reflux three times, and the extract was filtered and evaporated on a rotary evaporator under reduced pressure and then lyophilized. ACE was subjected to LC-MS/MS analysis and Scopoletin and liquiritin were found to be present at concentrations of 393.3±45.7 µg/g and 39.33±6.3 µg/g, respectively.

Cells

Huh7 and HepG2 human hepatocellular carcinoma cells were maintained in Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal bovine serum (FBS) and penicillin/streptomycin (100 U/ml). NIH3T3-W4P cell line expressing W4P mutant LHB was established and maintained as previously described (4).

Cell viability assay

Anti-proliferative effects of ACE on HCC cells were analyzed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Huh7 and HepG2 cells were treated with increasing concentrations of ACE for 72 h followed by further incubation with the MTT-containing medium for 4 h. After lysis with dimethyl sulfoxide, the absorbance of lysates was determined at 570 nm by using a microplate reader.

Colony forming assay

Six-hundred Huh7 cells and 200 cells of NIH3T3-W4P were seeded onto 6-well plates and incubated for 24 h. The NIH3T3 and Huh7 cells were then treated with different concentrations of ACE for 3 and 8 days, respectively. Colonies were fixed with 100% methanol for 10 min and then stained with 0.1% crystal violet for an hour. After washing with PBS, the number of colonies was counted.

Immunoblotting

The effect of ACE on the phosphorylation status of STAT3 was analyzed by immunoblotting. HepG2 cells were treated with ACE for 3 h prior to treatment with IL-6 (25 ng/ml). After incubation for 25 min, the cells were lysed and the cell lysates were subjected to immunoblotting using anti-STAT3 (Santa Cruz Biotechnology, sc-482), anti-phosphoSTAT3 (Cell Signaling, #9131), and anti-β-actin antibodies (Santa Cruz Biotechnology, sc-47778). To analyze the effect of ACE on the expression of cyclin D2 and P21, HepG2 cells were treated with ACE for 48 h and subjected to immunoblotting using anti-cyclin D2 (Santa Cruz Biotechnology, sc-593) and anti-P21 (Santa Cruz Biotechnology, I2807) antibodies.

RNA preparation and reverse-transcription quantitative PCR (RT-qPCR)

To determine the effect of ACE on the mRNA levels of cyclin D1, which are dependent on STAT3 transcriptional activity, RT-qPCR assays were performed. HepG2 cells and NIH3T3-W4P cells were treated with the increasing concentrations of ACE for 4 and 8 h, respectively. Total RNA was extracted using an RNeasy RNA extraction kit (Qiagen) according to the manufacturer's instructions. cDNA was synthesized from RNA using Superscript III reverse transcriptase (Invitrogen) with oligo20(dT) primers and analyzed by real-time PCR.

Scratch wound assay

Effect of ACE on Huh7 cell migration was investigated by performing a scratch wound assay. The confluent monolayer of Huh7 cells in a 6-well plate was wounded with a sterile pipette tip and incubated with increasing concentrations of ACE for 48 h. The cells were observed and images were obtained at 24 and 48 h after the treatment.

Transwell cell migration assay

Cell migration assay was performed as previously described (25). Briefly, after being treated with ACE (0, 25, 50, 100, and 200 µg/ml) for 24 h, Huh7 cells were harvested and seeded on a Boyden chamber (Neuro Probe, Cabin John, MD, USA) at a density of 105 cells per well in a serum-free medium and then incubated for 6 h at 37°C. The migrated cells were fixed with 100% methanol and stained with 5% Giemsa. Cells were counted under a light microscope.

Statistics

Statistical analysis was performed by SigmaPlot 10.0. Data are shown as means ± SD, and group differences were analyzed using paired Student's t-test. p<0.05 was considered as statistically significant.

Results

Anticancer effects of ACE on HCC cell lines

AC has been well known for its therapeutic effect in liver diseases and several previous studies suggested its antitumor activity. Here, we evaluated the anticancer activity of ACE on HCC by examining its effect on HCC cell proliferation by MTT assay. ACE displayed significant anti-proliferative effects against Huh7 and HepG2 human hepatoma cell lines in a dose-dependent manner (Fig. 1A and B). To further confirm the anticancer effect of ACE, its effect on the colony-forming ability of HCC cells was examined. As shown in Fig. 2, the colony formation of Huh7 cells was significantly inhibited in the presence of ACE; 100 µg/ml of ACE treatment resulted in 90% reduction of the colony formation. Compared to its direct cell cytotoxicity and the suppression of cell proliferation, ACE exerted potent inhibitory effect on colony forming.

Inhibition of STAT3 activation in HCC cell lines by ACE

Inhibition of proliferation and colony formation of HCC cells by ACE indicate its anticancer effect against HCC. Given that IL-6-mediated STAT3 activation is implicated in HCC development and progression, the role of ACE in STAT3 activation was examined. IL-6 treatment induced strong phosphorylation of STAT3, which is a hallmark of STAT3 activation in HepG2 cells. Treatment of IL-6-treated HepG2 cells with ACE resulted in marked decrease of both STAT3 and phosphorylated STAT3, indicating that ACE is capable of suppressing the action of STAT3 (Fig. 3A). Since STAT3 upregulates the transcription of cell cycle related genes including cyclin D1 and p21 and enhances cell cycle progression, the effect of ACE on the amounts of cyclin D1 and p21 was examined. Consistent with the suppression of STAT3 phosphorylation by ACE, synthesis of cyclin D1 mRNA as well as protein levels of cyclin D2 and p21 were also decreased by ACE treatment (Fig. 3B and C).

Inhibition of HBV W4P mutant large surface protein-induced STAT3 activation by ACE

In our previous study, we showed that male-specific W4P mutant HBV LHB drives HCC tumorigenesis by activating the IL-6/STAT3 signaling pathway (4). Thus, we evaluated whether ACE suppresses the STAT3 activation induced by W4P LHB. NIH3T3 cells stably expressing W4P LHB (NIH3T3-W4P) showed strong STAT3 phosphorylation compared to control NIH3T3 cells and the phosphorylation was decreased by ACE (Fig. 4A). The activity of ACE was dose-dependent and 100 µg/ml of ACE was sufficient to lower the phosphorylation to significant extent (Fig. 4B). In line with the results, cyclin D1 mRNA level of NIH3T3-W4P was also decreased in a dose-dependent manner by ACE treatment, further confirming that ACE suppresses the transcriptional activity of STAT3 (Fig. 4C). It has been shown that W4P LHB confers in vitro colony forming ability and in vivo tumor forming capability in nude mice to NIH3T3 cells by activating the STAT3 signaling pathway (4). Since ACE suppresses STAT3 activation in NIH3T3-W4P cells, we evaluated whether ACE suppresses the colony-forming capability of NIH3T3-W4P cells. As shown in Fig. 4D, ACE treatment decreased the number of colonies in a dose-dependent manner, and 250 µg/ml of ACE almost completely abolished the colony-forming ability of NIH3T3-W4P cells. These results suggest that ACE is capable of suppressing tumorigenesis induced by HBV mutations and subsequent inflammatory responses including STAT3 activation in addition to HCC cell proliferation.

Suppressive effects of ACE on HCC cell migration

STAT3 has been implicated in cancer cell migration and invasion through the upregulation of pro-migratory genes including MMP-2 (26). In the absence of ACE, Huh7 cells migrated along the edge and repaired approximately 30% of the wound after 48 h. Significant suppression of cell migration and wound recovery were observed in the presence of ACE and the inhibitory activity was concentration-dependent (Fig. 5A and B). Cell migration assay using Boyden chamber showed similar result. Treatment with 200 µg/ml of ACE suppressed over 95% of cell migration further confirming the suppressive activity of ACE on tumor cell migration (Fig. 6).

Discussion

AC has been widely used as a traditional remedy for various liver diseases in Asian countries. In the present study, we showed that ACE moderately suppressed the proliferation of HCC cells and it exerted stronger effect on colony-forming activity of HCC. In addition, we have shown that ACE treatment effectively suppressed STAT3 activation resulting in decreased cell proliferation and migration of HCC cells. Since its discovery, various tumor-promoting effects of STAT3, including cell cycle progression, anti-apoptosis, angiogenesis, and invasion/migration, have been studied and considered as key factors in oncogenesis and cancer progression (27). In particular, long-term IL-6-mediated inflammatory responses and subsequent STAT3 activation have been implicated in various liver pathogenic conditions including hepatitis, liver fibrosis, cirrhosis, and HCC carcinogenesis (28), suggesting that inhibition of STAT3 activation can be an effective strategy to treat various liver diseases. Given that IL-6/STAT3 signaling plays a crucial role in HCC development and progression, it is conceivable that the anticancer effect of ACE on HCC might be attributable to its inhibitory activity on the IL-6/STAT3 signaling pathway and subsequent cell cycle-related gene expression.

Chronic infection of HBV is a leading cause of HCC development and approximately 15–40% of chronic hepatitis B patients may develop progressed liver diseases including cirrhosis and HCC. In our previous study, we showed that a male-specific mutation of HBV is closely related to HCC development through the activation of STAT3. Ectopic expression of W4P LHB strongly induces STAT3 activation in NIH3T3 cells and transforming of the cells (4). In the current study, ACE suppressed not only STAT3 activation in IL-6-treated HCC cells but also STAT3 activation by W4P HBV LHB expression in fibroblast cells, suggesting that ACE may suppress tumorigenesis induced by HBV-mediated STAT3 activation. Thus, this study provides theoretical basis for the use of ACE in the treatment of HCC and prevention of HCC in chronic hepatitis patients.

The exact mechanisms of STAT3 inhibition by ACE and active ingredients inhibiting STAT3 activity remain elusive. Several compounds in ACE including capillarisin, scoparone, scopoletin and cholorogenic acid have been reported to suppress the activity of STAT3 (24,2931). Thus, it can be postulated that several compounds in ACE are responsible for STAT3 inhibition. ACE treatment on HepG2 cells resulted in decrease of STAT3 protein levels, indicating that the suppression of STAT3 activity is partly due to the lowered level of STAT3 (Fig. 3A). However, STAT3 phosphorylation of NIH3T3 cells induced by W4P LHB expression was markedly reduced despite the similar protein level of STAT3, indicating that ACE is also capable of suppressing the activation of STAT3 (Fig. 4A and B).

Regulation of IL-6/STAT3 signaling by ACE suggests its potential as a therapeutic agent against various liver diseases related to IL-6/STAT3 inflammatory responses. Identification of the constituents responsible for the STAT3 inhibition and unveiling of underlying molecular mechanism remain to be elucidated.

Acknowledgements

This study was supported by a grant of the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health and Welfare, Republic of Korea (grant number: HI14C0955).

References

1 

Abiru S, Migita K, Maeda Y, Daikoku M, Ito M, Ohata K, Nagaoka S, Matsumoto T, Takii Y, Kusumoto K, et al: Serum cytokine and soluble cytokine receptor levels in patients with non-alcoholic steatohepatitis. Liver Int. 26:39–45. 2006. View Article : Google Scholar : PubMed/NCBI

2 

Liu Y, Li P-K, Li C and Lin J: Inhibition of STAT3 signaling blocks the anti-apoptotic activity of IL-6 in human liver cancer cells. J Biol Chem. 285:27429–27439. 2010. View Article : Google Scholar : PubMed/NCBI

3 

Svinka J, Mikulits W and Eferl R: STAT3 in hepatocellular carcinoma: New perspectives. Hepat Oncol. 1:107–120. 2014. View Article : Google Scholar

4 

Lee SA, Kim H, Won YS, Seok SH, Na Y, Shin HB, Inn KS and Kim BJ: Male-specific hepatitis B virus large surface protein variant W4P potentiates tumorigenicity and induces gender disparity. Mol Cancer. 14:232015. View Article : Google Scholar : PubMed/NCBI

5 

Cronquist A: Vascular Flora of the Southeastern United States: Asteraceae. UNC Press Books. 2001.

6 

Nibret E and Wink M: Volatile components of four Ethiopian Artemisia species extracts and their in vitro antitrypanosomal and cytotoxic activities. Phytomedicine. 17:369–374. 2010. View Article : Google Scholar : PubMed/NCBI

7 

Han JM, Kim HG, Choi MK, Lee JS, Lee JS, Wang JH, Park HJ, Son SW, Hwang SY and Son CG: Artemisia capillaris extract protects against bile duct ligation-induced liver fibrosis in rats. Exp Toxicol Pathol. 65:837–844. 2013. View Article : Google Scholar : PubMed/NCBI

8 

Cha J-D, Jeong M-R, Jeong S-I, Moon SE, Kim JY, Kil BS and Song YH: Chemical composition and antimicrobial activity of the essential oils of Artemisia scoparia and A. capillaris. Planta Med. 71:186–190. 2005. View Article : Google Scholar

9 

Hong J-H, Hwang E-Y, Kim H-J, Jeong Y-J and Lee IS: Artemisia capillaris inhibits lipid accumulation in 3T3-L1 adipocytes and obesity in C57BL/6J mice fed a high fat diet. J Med Food. 12:736–745. 2009. View Article : Google Scholar : PubMed/NCBI

10 

Lee J, Chae K, Ha J, Park BY, Lee HS, Jeong S, Kim MY and Yoon M: Regulation of obesity and lipid disorders by herbal extracts from Morus alba, Melissa officinalis, and Artemisia capillaris in high-fat diet-induced obese mice. J Ethnopharmacol. 115:263–270. 2008. View Article : Google Scholar : PubMed/NCBI

11 

Mori H, Xu Q, Sakamoto O, Uesugi Y, Koda A and Nishioka I: Mechanisms of antitumor activity of aqueous extracts from Chinese herbs: Their immunopharmacological properties. Jpn J Pharmacol. 49:423–431. 1989. View Article : Google Scholar : PubMed/NCBI

12 

Hong JH and Lee IS: Effects of Artemisia capillaris ethyl acetate fraction on oxidative stress and antioxidant enzyme in high-fat diet induced obese mice. Chem Biol Interact. 179:88–93. 2009. View Article : Google Scholar : PubMed/NCBI

13 

Lee HI, Seo KO, Yun KW, Kim MJ and Lee MK: Comparative study of the hepatoprotective efficacy of Artemisia iwayomogi and Artemisia capillaris on ethanol-administered mice. J Food Sci. 76:T207–T211. 2011. View Article : Google Scholar : PubMed/NCBI

14 

Zhao Y, Geng CA, Ma YB, Huang XY, Chen H, Cao TW, He K, Wang H, Zhang XM and Chen JJ: UFLC/MS-IT-TOF guided isolation of anti-HBV active chlorogenic acid analogues from Artemisia capillaris as a traditional Chinese herb for the treatment of hepatitis. J Ethnopharmacol. 156:147–154. 2014. View Article : Google Scholar : PubMed/NCBI

15 

Zhao Y, Geng CA, Chen H, Ma YB, Huang XY, Cao TW, He K, Wang H, Zhang XM and Chen JJ: Isolation, synthesis and anti-hepatitis B virus evaluation of p-hydroxyacetophenone derivatives from Artemisia capillaris. Bioorg Med Chem Lett. 25:1509–1514. 2015. View Article : Google Scholar : PubMed/NCBI

16 

Zhao Y, Geng CA, Sun CL, Ma YB, Huang XY, Cao TW, He K, Wang H, Zhang XM and Chen JJ: Polyacetylenes and anti-hepatitis B virus active constituents from Artemisia capillaris. Fitoterapia. 95:187–193. 2014. View Article : Google Scholar : PubMed/NCBI

17 

Kim KS, Yang HJ, Lee JY, Na YC, Kwon SY, Kim YC, Lee JH and Jang HJ: Effects of β-sitosterol derived from Artemisia capillaris on the activated human hepatic stellate cells and dimethylnitrosamine-induced mouse liver fibrosis. BMC Complement Altern Med. 14:3632014. View Article : Google Scholar : PubMed/NCBI

18 

Wang J-H, Choi M-K, Shin J-W, Hwang S-Y and Son C-G: Antifibrotic effects of Artemisia capillaris and Artemisia iwayomogi in a carbon tetrachloride-induced chronic hepatic fibrosis animal model. J Ethnopharmacol. 140:179–185. 2012. View Article : Google Scholar : PubMed/NCBI

19 

Altekruse SF, McGlynn KA and Reichman ME: Hepatocellular carcinoma incidence, mortality, and survival trends in the United States from 1975 to 2005. J Clin Oncol. 27:1485–1491. 2009. View Article : Google Scholar : PubMed/NCBI

20 

Thomas MB and Zhu AX: Hepatocellular carcinoma: The need for progress. J Clin Oncol. 23:2892–2899. 2005. View Article : Google Scholar : PubMed/NCBI

21 

Yang C-C, Lee M-R, Hsu S-L and Chang C-MJ: Supercritical fluids extraction of capillarisin from Artemisia capillaris and its inhibition of in vitro growth of hepatoma cells. J Supercrit Fluids. 42:96–103. 2007. View Article : Google Scholar

22 

Hu YQ, Tan RX, Chu MY and Zhou J: Apoptosis in human hepatoma cell line SMMC-7721 induced by water-soluble macromolecular components of Artemisia capillaris Thunberg. Jpn J Cancer Res. 91:113–117. 2000. View Article : Google Scholar : PubMed/NCBI

23 

Hong SH, Seo SH, Lee JH and Choi BT: The aqueous extract from Artemisia capillaris Thunb. inhibits lipopolysaccharide-induced inflammatory response through preventing NF-kappaB activation in human hepatoma cell line and rat liver. Int J Mol Med. 13:717–720. 2004.

24 

Lee JH, Chiang SY, Nam D, Chung WS, Lee J, Na YS, Sethi G and Ahn KS: Capillarisin inhibits constitutive and inducible STAT3 activation through induction of SHP-1 and SHP-2 tyrosine phosphatases. Cancer Lett. 345:140–148. 2014. View Article : Google Scholar : PubMed/NCBI

25 

Yang SF, Chen MK, Hsieh YS, Yang JS, Zavras AI, Hsieh YH, Su SC, Kao TY, Chen PN and Chu SC: Antimetastatic effects of Terminalia catappa L. on oral cancer via a down-regulation of metastasis-associated proteases. Food Chem Toxicol. 48:1052–1058. 2010.

26 

Aggarwal BB, Kunnumakkara AB, Harikumar KB, Gupta SR, Tharakan ST, Koca C, Dey S and Sung B: Signal transducer and activator of transcription-3, inflammation, and cancer: How intimate is the relationship? Ann NY Acad Sci. 1171:59–76. 2009. View Article : Google Scholar : PubMed/NCBI

27 

Furtek SL, Backos DS, Matheson CJ and Reigan P: Strategies and approaches of targeting STAT3 for cancer treatment. ACS Chem Biol. 11:308–318. 2016. View Article : Google Scholar : PubMed/NCBI

28 

Subramaniam A, Shanmugam MK, Perumal E, Li F, Nachiyappan A, Dai X, Swamy SN, Ahn KS, Kumar AP, Tan BK, et al: Potential role of signal transducer and activator of transcription (STAT)3 signaling pathway in inflammation, survival, proliferation and invasion of hepatocellular carcinoma. Biochim Biophys Acta. 1835:46–60. 2013.PubMed/NCBI

29 

Kim JK, Kim JY, Kim HJ, Park KG, Harris RA, Cho WJ, Lee JT and Lee IK: Scoparone exerts anti-tumor activity against DU145 prostate cancer cells via inhibition of STAT3 activity. PLoS One. 8:e803912013. View Article : Google Scholar : PubMed/NCBI

30 

Bhattacharyya SS, Paul S, Dutta S, Boujedaini N and Khuda-Bukhsh AR: Anti-oncogenic potentials of a plant coumarin (7-hydroxy-6-methoxy coumarin) against 7,12-dimethylbenz [a] anthracene-induced skin papilloma in mice: The possible role of several key signal proteins. Zhong Xi Yi Jie He Xue Bao. 8:645–654. 2010. View Article : Google Scholar : PubMed/NCBI

31 

Tan Z, Luo M, Yang J, Cheng Y, Huang J, Lu C, Song D, Ye M, Dai M, Gonzalez FJ, et al: Chlorogenic acid inhibits cholestatic liver injury induced by α-naphthylisothiocyanate: Involvement of STAT3 and NFκB signalling regulation. J Pharm Pharmacol. 68:1203–1213. 2016. View Article : Google Scholar : PubMed/NCBI

Related Articles

Journal Cover

January-2017
Volume 37 Issue 1

Print ISSN: 1021-335X
Online ISSN:1791-2431

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
x
Spandidos Publications style
Jang E, Kim S, Lee N, Yi C, Hong D, Lee WS, Kim J, Lee K, Kim B, Lee J, Lee J, et al: Evaluation of antitumor activity of Artemisia capillaris extract against hepatocellular carcinoma through the inhibition of IL-6/STAT3 signaling axis. Oncol Rep 37: 526-532, 2017
APA
Jang, E., Kim, S., Lee, N., Yi, C., Hong, D., Lee, W.S. ... Inn, K. (2017). Evaluation of antitumor activity of Artemisia capillaris extract against hepatocellular carcinoma through the inhibition of IL-6/STAT3 signaling axis. Oncology Reports, 37, 526-532. https://doi.org/10.3892/or.2016.5283
MLA
Jang, E., Kim, S., Lee, N., Yi, C., Hong, D., Lee, W. S., Kim, J., Lee, K., Kim, B., Lee, J., Inn, K."Evaluation of antitumor activity of Artemisia capillaris extract against hepatocellular carcinoma through the inhibition of IL-6/STAT3 signaling axis". Oncology Reports 37.1 (2017): 526-532.
Chicago
Jang, E., Kim, S., Lee, N., Yi, C., Hong, D., Lee, W. S., Kim, J., Lee, K., Kim, B., Lee, J., Inn, K."Evaluation of antitumor activity of Artemisia capillaris extract against hepatocellular carcinoma through the inhibition of IL-6/STAT3 signaling axis". Oncology Reports 37, no. 1 (2017): 526-532. https://doi.org/10.3892/or.2016.5283