Liquiritin inhibits proliferation and induces apoptosis in HepG2 hepatocellular carcinoma cells via the ROS-mediated MAPK/AKT/NF-κB signaling pathway

Wang, Jia-Ru; Li, Tian-Zhu; Wang, Cheng; Li, Shu-Mei; Luo, Ying-Hua; Piao, Xian-Ji; Feng, Yu-Chao; Zhang, Yi; Xu, Wan-Ting; Zhang, Yu; Zhang, Tong; Wang, Shi-Nong; Xue, Hui; Wang, Hong-Xing; Cao, Long-Kui; Jin, Cheng-Hao

doi:10.1007/s00210-019-01763-7

Original Article
Published: 20 January 2020

Liquiritin inhibits proliferation and induces apoptosis in HepG2 hepatocellular carcinoma cells via the ROS-mediated MAPK/AKT/NF-κB signaling pathway

Jia-Ru Wang¹,
Tian-Zhu Li²,
Cheng Wang³,
Shu-Mei Li⁴,
Ying-Hua Luo⁵,
Xian-Ji Piao⁶,
Yu-Chao Feng⁷,
Yi Zhang¹,
Wan-Ting Xu¹,
Yu Zhang¹,
Tong Zhang¹,
Shi-Nong Wang¹,
Hui Xue¹,
Hong-Xing Wang¹,
Long-Kui Cao^7,8 &
Cheng-Hao Jin ORCID: orcid.org/0000-0003-4431-2623^1,7,8

Naunyn-Schmiedeberg's Archives of Pharmacology volume 393, pages 1987–1999 (2020)Cite this article

559 Accesses
9 Citations
Metrics details

Abstract

Liquiritin (LIQ), a major constituent of Glycyrrhiza Radix, exhibits various pharmacological activities. In this study, to explore the potential anti-cancer effects and its underlying molecular mechanisms of LIQ in hepatocellular carcinoma (HCC) cells. LIQ significantly decreased viability and induced apoptosis in HepG2 cells by decreasing mitochondrial membrane potential and regulating Bcl-2 family proteins, cytochrome c, cle-caspase-3, and cle-PARP. The cell cycle analysis and western blot analysis revealed that LIQ induced G2/M phase arrest through increased expression of p21 and decreased levels of p27, cyclin B, and CDK1/2. The flow cytometry and western blot analysis also suggested that LIQ promoted the accumulation of ROS in HepG2 cells and up-regulated the phosphorylation expression levels of p38 kinase, c-Jun N-terminal kinase (JNK), and inhibitor of NF-κB (IκB-α); the phosphorylation levels of extracellular signal-regulated kinase (ERK), protein kinase B (AKT), signal transducer activator of transcription 3 (STAT3), and nuclear factor kappa B (NF-κB) were down-regulated. However, these effects were reversed by N-acetyl-l-cysteine (NAC), MAPK, and AKT inhibitors. The findings demonstrated that LIQ induced cell cycle arrest and apoptosis via the ROS-mediated MAPK/AKT/NF-κB signaling pathway in HepG2 cells, and the LIQ may serve as a potential therapeutic agent for the treatment of human HCC.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

USD 39.95

Price includes VAT (USA)
Tax calculation will be finalised during checkout.

References

Belayachi L, Aceves-Luquero C, Merghoub N, de Mattos SF, Amzazi S, Villalonga P, Bakri Y (2017) Induction of cell cycle arrest and apoptosis by ormenis eriolepis a morrocan endemic plant in various human cancer cell lines. Afr J Tradit Complement Altern Med 14:356–373. https://doi.org/10.21010/ajtcam.v14i2.37
CAS Article PubMed PubMed Central Google Scholar
Bours V, Bentires-Alj M, Hellin AC, Viatour P, Robe P, Delhalle S, Benoit V, Merville MP (2000) Nuclear factor-kappa B, cancer, and apoptosis. Biochem Pharmacol 60:1085–1089
CAS Article PubMed Google Scholar
Chen H, Zhou L, Wu X, Li R, Wen J, Sha J, Wen X (2016) The PI3K/AKT pathway in the pathogenesis of prostate cancer. Front Biosci (Landmark Ed) 21:1084–1091
CAS Article Google Scholar
Chetram MA, Bethea DA, Odero-Marah VA, Don-Salu-Hewage AS, Jones KJ, Hinton CV (2013) ROS-mediated activation of AKT induces apoptosis via pVHL in prostate cancer cells. Mol Cell Biochem 376:63–71. https://doi.org/10.1007/s11010-012-1549-7
CAS Article PubMed PubMed Central Google Scholar
Cui YQ, Liu YJ, Zhang F (2018) The suppressive effects of Britannin (Bri) on human liver cancer through inducing apoptosis and autophagy via AMPK activation regulated by ROS. Biochem Biophys Res Commun 497:916–923. https://doi.org/10.1016/j.bbrc.2017.12.144
CAS Article PubMed Google Scholar
Eruslanov E, Kusmartsev S (2010) Identification of ROS using oxidized DCFDA and flow-cytometry. Methods Mol Biol 594:57–72. https://doi.org/10.1007/978-1-60761-411-1_4
CAS Article PubMed Google Scholar
Gierasch LM (2016) The Journal of Biological Chemistry: 2016 Onward. J Biol Chem 291(27):15406–15407. https://doi.org/10.1074/jbc.E116.000001
Article PubMed PubMed Central Google Scholar
Goldar S, Khaniani MS, Derakhshan SM, Baradaran B (2015) Molecular mechanisms of apoptosis and roles in cancer development and treatment. Asian Pac J Cancer Prev 16:2129–2144
Article PubMed Google Scholar
Gorrini C, Harris IS, Mak TW (2013) Modulation of oxidative stress as an anticancer strategy. Nat Rev Drug Discov 12(12):931–947. https://doi.org/10.1038/nrd4002
CAS Article PubMed Google Scholar
Hao W, Yuan X, Yu L, Gao C, Sun X, Wang D, Zheng Q (2015) Licochalcone A-induced human gastric cancer BGC-823 cells apoptosis by regulating ROS-mediated MAPKs and PI3K/AKT signaling pathways. Sci Rep 5:10336–10343. https://doi.org/10.1038/srep10336
CAS Article PubMed PubMed Central Google Scholar
He SH, Liu HG, Zhou YF, Yue QF (2017) Liquiritin (LT) exhibits suppressive effects against the growth of human cervical cancer cells through activating Caspase-3 in vitro and xenograft mice in vivo. Biomed Pharmacother 92:215–228. https://doi.org/10.1016/j.biopha.2017.05.026
CAS Article PubMed Google Scholar
Ke F, Wang Z, Song X, Ma Q, Hu Y, Jiang L, Zhang Y, Liu Y, Zhang Y, Gong W (2017) Cryptotanshinone induces cell cycle arrest and apoptosis through the JAK2/STAT3 and PI3K/Akt/NFκB pathways in cholangiocarcinoma cells. Drug Des Devel Ther 11:1753–1766. https://doi.org/10.2147/DDDT.S132488
CAS Article PubMed PubMed Central Google Scholar
Ku JM, Kim SR, Hong SH, Choi HS, Seo HS, Shin YC, Ko SG (2015) Cucurbitacin D induces cell cycle arrest and apoptosis by inhibiting STAT3 and NF-κB signaling in doxorubicin-resistant human breast carcinoma (MCF7/ADR) cells. Mol Cell Biochem 409:33–43. https://doi.org/10.1007/s11010-015-2509-9
CAS Article PubMed PubMed Central Google Scholar
Li XQ, Cai LM, Liu J, Ma YL, Kong YH, Li H, Jiang (2018) Liquiritin suppresses UVB-induced skin injury through prevention of inflammation, oxidative stress and apoptosis through the TLR4/MyD88/NF-κB and MAPK/caspase signaling pathways. Int J Mol Med 42:1445-1459. https://doi.org/10.3892/ijmm.2018.3720.
Lin MT, Lin CL, Lin TY, Cheng CW, Yang SF, Lin CL, Wu CC, Hsieh YH, Tsai JP (2016) Synergistic effect of fisetin combined with sorafenib in human cervical cancer HeLa cells through activation of death receptor-5 mediated caspase-8/caspase-3 and the mitochondria-dependent apoptotic pathway. Tumour Biol 37:6987–6996. https://doi.org/10.1007/s13277-015-4526-4
CAS Article PubMed Google Scholar
Liu CY, Chen KF, Chen PJ (2015) Treatment of liver cancer. Cold Spring Harb Perspect Med 5:a021535. https://doi.org/10.1101/cshperspect.a021535
CAS Article PubMed PubMed Central Google Scholar
Marzo I, Naval J (2008) Bcl-2 family members as molecular targets in cancer therapy. Biochem Pharmacol 76:939–946. https://doi.org/10.1016/j.bcp.2008.06.009
CAS Article PubMed Google Scholar
Ola MS, Nawaz M, Ahsan H (2011) Role of Bcl-2 family proteins and caspases in the regulation of apoptosis. Mol Cell Biochem 351:41–58. https://doi.org/10.1007/s11010-010-0709-x
CAS Article PubMed Google Scholar
Ozer Etik D, Suna N, Boyacioglu AS (2017) Management of hepatocellular carcinoma: prevention, surveillance, diagnosis, and staging. Exp Clin Transplant 15:31–35. https://doi.org/10.6002/ect.TOND16.L9
Article PubMed Google Scholar
Palanivel K, Kanimozhi V, Kadalmani B, Akbarsha MA (2014) Verrucarin A induces apoptosis through ROS-mediated EGFR/MAPK/Akt signaling pathways in MDA-MB-231 breast cancer cells. J Cell Biochem 115:2022–2032. https://doi.org/10.1002/jcb.24874
CAS Article PubMed Google Scholar
Puthdee N, Seubwai W, Vaeteewoottacharn K, Boonmars T, Cha’on U, Phoomak C, Wongkham S (2017) Berberine induces cell cycle arrest in cholangiocarcinoma cell lines via inhibition of NF-κB and STAT3 pathways. Biol Pharm Bull 40:751–757. https://doi.org/10.1248/bpb.b16-00428
CAS Article PubMed Google Scholar
Rawlings JS, Rosler KM, Harrison DA (2004) The JAK/STAT signaling pathway. J Cell Sci 117:1281–1283. https://doi.org/10.1242/jcs.00963
CAS Article PubMed Google Scholar
Shao D, Ma J, Zhou C, Zhao JN, Li LL, Zhao TJ, Ai XL, Jiao P (2017) STAT3 down-regulation induces mitochondria-dependent G2/M cell cycle arrest and apoptosis in oesophageal carcinoma cells. Clin Exp Pharmacol Physiol 44:413–420. https://doi.org/10.1111/1440-1681.12708
CAS Article PubMed Google Scholar
Sun SY, Hail N Jr, Lotan R (2004) Apoptosis as a novel target for cancer chemoprevention. J Natl Cancer Inst 96:662–672
CAS Article PubMed Google Scholar
Sun DK, Wang L, Zhang P (2017) Antitumor effects of chrysanthemin in PC-3 human prostate cancer cells are mediated via apoptosis induction, caspase signalling pathway and loss of mitochondrial membrane potential. Afr J Tradit Complement Altern Med. 14:54–61. https://doi.org/10.21010/ajtcam.v14i4.7
CAS Article PubMed PubMed Central Google Scholar
Tobiume K, Matsuzawa A, Takahashi T, Nishitoh H, Morita K, Takeda K, Minowa O, Miyazono K, Noda T, Ichijo H (2001) ASK1 is required for sustained activations of JNK/p38 MAP kinases and apoptosis. EMBO Rep 2(3):222–228
CAS Article PubMed PubMed Central Google Scholar
Tsai WC, Kung PT, Wang YH, Kuo WY, Li YH (2018) Influence of the time interval from diagnosis to treatment on survival for early-stage liver cancer. PLoS One 13:e0199532. https://doi.org/10.1371/journal.pone.0199532
CAS Article PubMed PubMed Central Google Scholar
Wallace MC, Preen D, Jeffrey GP, Adams LA (2015) The evolving epidemiology of hepatocellular carcinoma: a global perspective. Expert Rev Gastroenterol Hepatol 9:765–779. https://doi.org/10.1586/17474124.2015.1028363
CAS Article PubMed Google Scholar
Wang D, Lu J, Liu Y, Meng Q, Xie J, Wang Z, Teng L (2014) Liquiritigenin induces tumor cell death through mitogen-activated protein kinase- (MPAKs-) mediated pathway in hepatocellular carcinoma cells. Biomed Res Int 2014:965316–965326. https://doi.org/10.1155/2014/965316
CAS Article PubMed PubMed Central Google Scholar
Wang ST, Ho HJ, Lin JT, Shieh JJ, Wu CY (2017) Simvastatin-induced cell cycle arrest through inhibition of STAT3/SKP2 axis and activation of AMPK to promote p27 and p21 accumulation in hepatocellular carcinoma cells. Cell Death Dis 8:e2626. https://doi.org/10.1038/cddis.2016.472
CAS Article PubMed PubMed Central Google Scholar
Wang JR, Luo YH, Piao XJ, Zhang Y, Feng YC, Li JQ, Xu WT, Zhang Y, Zhang T, Wang SN, Xue H, Wang WZ, Cao LK, Jin CH (2019) Mechanisms underlying isoliquiritigenin-induced apoptosis and cell cycle arrest via ROS-mediated MAPK/STAT3/NF-κB pathways in human hepatocellular carcinoma cells. Drug Dev Res 80(4):461–470. https://doi.org/10.1002/ddr.21518
CAS Article PubMed Google Scholar
Wei F, Jiang X, Gao HY, Gao SH (2017) Liquiritin induces apoptosis and autophagy in cisplatin (DDP)-resistant gastric cancer cells in vitro and xenograft nude mice in vivo. Int J Oncol 51:1383–1394. https://doi.org/10.3892/ijo.2017.4134
CAS Article PubMed PubMed Central Google Scholar
Wong RS (2011) Apoptosis in cancer: from pathogenesis to treatment. J Exp Clin Cancer Res 30:87–100. https://doi.org/10.1186/1756-9966-30-87
CAS Article PubMed PubMed Central Google Scholar
Xiang Y, Ye W, Huang C, Lou B, Zhang J, Yu D, Huang X, Chen B, Zhou M (2017) Brusatol inhibits growth and induces apoptosis in pancreatic cancer cells via JNK/p38 MAPK/NF-κb/Stat3/Bcl-2 signaling pathway. Biochem Biophys Res Commun 487:820–826. https://doi.org/10.1016/j.bbrc.2017.04.133
CAS Article PubMed Google Scholar
Xu W, Mi Y, He P, Sheng H, Ling N (2017) γ-Tocotrienol inhibits proliferation and induces apoptosis via the mitochondrial pathway in human cervical cancer HeLa cells. Molecules 22:1299–1303. https://doi.org/10.3390/molecules22081299
CAS Article PubMed Central Google Scholar
Ye C, Zhao W, Li M, Zhuang J, Yan X, Lu Q, Chang C, Huang X, Zhou J, Xie B, Zhang Z, Yao X, Yan J, Guo H (2015) Lipid raft localization of epidermal growth factor receptor alters matrix metalloproteinase-1 expression in SiHa cells via the MAPK/ERK signaling pathway. Oncol Lett 12:4991–4998. https://doi.org/10.3892/ol.2016.5307
CAS Article Google Scholar
Yu CY, Jerry-Teng CL, Hung PS, Cheng CC, Hsu SL, Hwang GY, Tzeng YM (2018) Ovatodiolide isolated from Anisomeles indica induces cell cycle G2/M arrest and apoptosis via a ROS-dependent ATM/ATR signaling pathways. Eur J Pharmacol 819:16–29. https://doi.org/10.1016/j.ejphar.2017.09.050
CAS Article PubMed Google Scholar
Zhong WF, Wang XH, Pan B, Li F, Kuang L, Su ZX (2016) Eupatilin induces human renal cancer cell apoptosis via ROS-mediated MAPK and PI3K/AKT signaling pathways. Oncol Lett 12:2894–2899. https://doi.org/10.3892/ol.2016.4989
CAS Article PubMed PubMed Central Google Scholar
Zhou Y, Ho WS (2014) Combination of liquiritin, isoliquiritin and isoliquirigenin induce apoptotic cell death through upregulating p53 and p21 in the A549 non-small cell lung cancer cells. Oncol Rep 31:298–304. https://doi.org/10.3892/or.2013.2849
CAS Article PubMed Google Scholar
Zhou D, Zhou Y, Li C, Yang L (2018) Silencing of B7-H4 suppresses the tumorigenicity of the MGC-803 human gastric cancer cell line and promotes cell apoptosis via the mitochondrial signaling pathway. Int J Oncol 52:1267–1276. https://doi.org/10.3892/ijo.2018.4274
CAS Article PubMed Google Scholar
Zhu Y, Jiang Y, Shi L, Du L, Xu X, Wang E, Sun Y, Guo X, Zou B, Wang H, Wang C, Sun L, Zhen Y (2017) 7-O-Geranylquercetin induces apoptosis in gastric cancer cells via ROS-MAPK mediated mitochondrial signaling pathway activation. Biomed Pharmacother 87:527–538. https://doi.org/10.1016/j.biopha.2016.12.095
CAS Article PubMed Google Scholar

Download references

Funding

This work was funded by the Postdoctoral Scientific Research Foundation of Heilongjiang Province of China (No. LBH-Q13132), Heilongjiang Touyan Innovation Team Program (No.2019HTY078), “Regulation and activity retention technology and application of coarse cereals active components in processing” (No. 2017YFD0401203), and the Heilongjiang Farms & Land Reclamation Administration Support Project for Key Scientific Research (No. HKKYZD190705), The Heilongjiang Bayi Agricultural University Support program for ‘San Zong’(No.TDJH201905).

Author information

Jia-Ru Wang, Tian-Zhu Li, and Cheng Wang contributed equally to this work.

Authors and Affiliations

Department of Biochemistry and Molecular Biology, College of Life Science & Technology, Heilongjiang Bayi Agricultural University, Daqing, 163319, China
Jia-Ru Wang, Yi Zhang, Wan-Ting Xu, Yu Zhang, Tong Zhang, Shi-Nong Wang, Hui Xue, Hong-Xing Wang & Cheng-Hao Jin
Molecular Medicine Research Center, School of Basic Medical Science, Chifeng University, Chifeng, 024000, China
Tian-Zhu Li
Pharmacy Department, Daqing Oilfield General Hospital, Daqing, 163001, China
Cheng Wang
Hemodialysis Center, Daqing Oilfield General Hospital, Daqing, 163001, China
Shu-Mei Li
Department of Grass Science, College of Animal Science & Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing, 163319, China
Ying-Hua Luo
Department of Gynaecology and Obstetrics, the Fifth Affiliated Hospital of Harbin Medical University, Daqing, 163316, China
Xian-Ji Piao
Department of Food Science and Engineering, College of Food Science & Technology, Heilongjiang Bayi Agricultural University, Daqing, 163319, China
Yu-Chao Feng, Long-Kui Cao & Cheng-Hao Jin
National Coarse Cereals Engineering Research Center, Daqing, 163319, Heilongjiang, China
Long-Kui Cao & Cheng-Hao Jin

Authors

Jia-Ru Wang
View author publications
You can also search for this author in PubMed Google Scholar
Tian-Zhu Li
View author publications
You can also search for this author in PubMed Google Scholar
Cheng Wang
View author publications
You can also search for this author in PubMed Google Scholar
Shu-Mei Li
View author publications
You can also search for this author in PubMed Google Scholar
Ying-Hua Luo
View author publications
You can also search for this author in PubMed Google Scholar
Xian-Ji Piao
View author publications
You can also search for this author in PubMed Google Scholar
Yu-Chao Feng
View author publications
You can also search for this author in PubMed Google Scholar
Yi Zhang
View author publications
You can also search for this author in PubMed Google Scholar
Wan-Ting Xu
View author publications
You can also search for this author in PubMed Google Scholar
Yu Zhang
View author publications
You can also search for this author in PubMed Google Scholar
Tong Zhang
View author publications
You can also search for this author in PubMed Google Scholar
Shi-Nong Wang
View author publications
You can also search for this author in PubMed Google Scholar
Hui Xue
View author publications
You can also search for this author in PubMed Google Scholar
Hong-Xing Wang
View author publications
You can also search for this author in PubMed Google Scholar
Long-Kui Cao
View author publications
You can also search for this author in PubMed Google Scholar
Cheng-Hao Jin
View author publications
You can also search for this author in PubMed Google Scholar

Contributions

CHJ and LKC conceived and designed the study. JR W and TZL wrote the paper and performed some of the experiments. CW, YHL, and XJP assessed the cytotoxic effects. YCF, YZ, and WTX performed the apoptotic analyses. YZ and TZ performed the cell cycle analyses. SML, SNW, and HX performed the western blot analyses. HXW performed the signaling analyses. All of the authors have read and approved the final version of the manuscript.

Corresponding authors

Correspondence to Long-Kui Cao or Cheng-Hao Jin.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Supplemental Fig. 1

(DOCX 569 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Wang, JR., Li, TZ., Wang, C. et al. Liquiritin inhibits proliferation and induces apoptosis in HepG2 hepatocellular carcinoma cells via the ROS-mediated MAPK/AKT/NF-κB signaling pathway. Naunyn-Schmiedeberg's Arch Pharmacol 393, 1987–1999 (2020). https://doi.org/10.1007/s00210-019-01763-7

Download citation

Received: 21 July 2019
Accepted: 01 November 2019
Published: 20 January 2020
Issue Date: October 2020
DOI: https://doi.org/10.1007/s00210-019-01763-7

Keywords

Liquiritin
Human hepatocellular carcinoma cells
Apoptosis
Cell cycle arrest
Reactive oxygen species