Herbal Formula YGJDSJ Inhibits Anchorage-independent Growth and Induces Anoikis in Hepatocellular Carcinoma Bel-7402 Cells

Overview

Journal BMC Complement Altern Med

Publisher Biomed Central

Specialty Rehabilitation Medicine

Date 2018 Jan 18

PMID 29338725

Citations 5

Authors

Bing Hu

Tong Zhang

Hong-Mei An

Jia-Lu Zheng

Xia Yan

Xiao-Wei Huang

Affiliations

Soon will be listed here.

Abstract

Background: Based on clinical medications and related studies, we established a Yang-Gan Jie-Du Sang-Jie (YGJDSJ) herbal formula for hepatocarcinoma treatment. In present study, we evaluated the anti-cancer potential of YGJDSJ on suspension-grown human hepatocellular carcinoma Bel-7402 cells.

Methods: Bel-7402 cells were cultured in poly(2-hydroxyethyl methacrylate) (poly-HEMA) coated plates and treated with YGJDSJ. Anchorage-independent cell growth was detected by cell Counting Kit-8 (CCK-8) assay and soft agar colony formation assay. Anoikis was detected by ethdium homodimer-1 (EthD-1) staining and flow cytometry analysis. Caspases activities were detected by the cleavage of chromogenic substrate. Reactive oxygen species (ROS) was detected by 2',7'-dichlorofluorescin diacetate (DCFH-DA) staining. Protein expression and phosphorylation was identified by western blot. Protein expression was knocked-down by siRNA.

Results: YGJDSJ inhibited the proliferation of Bel-7402 cells in poly-HEMA coated plates and anchorage-independent growth of Bel-7402 cells in soft agar. YGJDSJ also induced anoikis in Bel-7402 cells as indicated by EthD-1 staining and flow cytometry analysis. YGJDSJ activated caspase-3, - 8, and - 9 in suspension-grown Bel-7402 cells. The pan-caspase inhibitor Z-VAD-FMK significantly abrogated the effects of YGJDSJ on anoikis in suspension-grown Bel-7402 cells. In addition, YGJDSJ increased ROS in suspension-grown Bel-7402 cells. The ROS scavenger N-acetyl-L-cysteine (NAC) partially attenuated YGJDSJ-induced activation of caspase-3, - 8 and - 9 and anoikis in suspension-grown Bel-7402 cells. Furthermore, YGJDSJ inhibited expression and phosphorylation of protein tyrosine kinase 2 (PTK2) in suspension-grown Bel-7402 cells. Over-expression of PTK2 significantly abrogated YGJDSJ induced anoikis.

Conclusions: YGJDSJ inhibits anchorage-independent growth and induce caspase-mediated anoikis in Bel-7402 cells, and may relate to ROS generation and PTK2 downregulation.

Citing Articles

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Adeshakin F, Adeshakin A, Afolabi L, Yan D, Zhang G, Wan X Front Oncol. 2021; 11:626577.

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Chen F, Zhong Z, Tan H, Guo W, Zhang C, Tan C Front Pharmacol. 2020; 11:293.

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Traditional Chinese medicine formulation Yanggan Jiedu Sanjie inhibits TGF-β1-induced epithelial-mesenchymal transition and metastatic potential in human hepatocarcinoma Bel-7402 cells.

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References

Cheng J, Han W, Wang Z, Shao Y, Wang Y, Zhang Y . Hepatocellular Carcinoma Growth Is Inhibited by Euphorbia helioscopia L. Extract in Nude Mice Xenografts. Biomed Res Int. 2015; 2015:601015. PMC: 4450285. DOI: 10.1155/2015/601015. View

Thiyagarajan V, Lin S, Lee C, Weng C . A focal adhesion kinase inhibitor 16-hydroxy-cleroda-3,13-dien-16,15-olide incorporated into enteric-coated nanoparticles for controlled anti-glioma drug delivery. Colloids Surf B Biointerfaces. 2016; 141:120-131. DOI: 10.1016/j.colsurfb.2016.01.038. View

Lau A, Wang Y, Chiu J . Reactive oxygen species: current knowledge and applications in cancer research and therapeutic. J Cell Biochem. 2008; 104(2):657-67. DOI: 10.1002/jcb.21655. View

Sakamoto S, Kyprianou N . Targeting anoikis resistance in prostate cancer metastasis. Mol Aspects Med. 2010; 31(2):205-14. PMC: 2988681. DOI: 10.1016/j.mam.2010.02.001. View

Wang X, Jiang Y, Hu D . Optimization and in vitro antiproliferation of Curcuma wenyujin's active extracts by ultrasonication and response surface methodology. Chem Cent J. 2016; 10:32. PMC: 4868111. DOI: 10.1186/s13065-016-0177-9. View

Paoli P, Giannoni E, Chiarugi P . Anoikis molecular pathways and its role in cancer progression. Biochim Biophys Acta. 2013; 1833(12):3481-3498. DOI: 10.1016/j.bbamcr.2013.06.026. View

Coates J, Galante J, Bold R . Cancer therapy beyond apoptosis: autophagy and anoikis as mechanisms of cell death. J Surg Res. 2009; 164(2):301-8. DOI: 10.1016/j.jss.2009.07.011. View

Yang S, Wang L, Kong Q . Depression of focal adhesion kinase induces apoptosis in rat osteosarcoma OSR-6 cells in a caspase-dependent pathway. Cell Biochem Biophys. 2014; 70(2):765-70. DOI: 10.1007/s12013-014-9979-3. View

Terasaki M, Maeda H, Miyashita K, Mutoh M . Induction of Anoikis in Human Colorectal Cancer Cells by Fucoxanthinol. Nutr Cancer. 2017; 69(7):1043-1052. DOI: 10.1080/01635581.2017.1339814. View

10.

Hu B, Shen K, An H, Wu Y, Du Q . Aqueous extract of Curcuma aromatica induces apoptosis and G2/M arrest in human colon carcinoma LS-174-T cells independent of p53. Cancer Biother Radiopharm. 2011; 26(1):97-104. DOI: 10.1089/cbr.2010.0853. View

11.

Duxbury M, Ito H, Zinner M, Ashley S, Whang E . Focal adhesion kinase gene silencing promotes anoikis and suppresses metastasis of human pancreatic adenocarcinoma cells. Surgery. 2004; 135(5):555-62. DOI: 10.1016/j.surg.2003.10.017. View

12.

Duseja A . Staging of hepatocellular carcinoma. J Clin Exp Hepatol. 2015; 4(Suppl 3):S74-9. PMC: 4284240. DOI: 10.1016/j.jceh.2014.03.045. View

13.

Hu B, Du Q, Deng S, An H, Pan C, Shen K . Ligustrum lucidum Ait. fruit extract induces apoptosis and cell senescence in human hepatocellular carcinoma cells through upregulation of p21. Oncol Rep. 2014; 32(3):1037-42. DOI: 10.3892/or.2014.3312. View

14.

Lieman J, Worley L, Harbour J . Loss of Rb-E2F repression results in caspase-8-mediated apoptosis through inactivation of focal adhesion kinase. J Biol Chem. 2005; 280(11):10484-90. DOI: 10.1074/jbc.M409371200. View

15.

Circu M, Aw T . Reactive oxygen species, cellular redox systems, and apoptosis. Free Radic Biol Med. 2010; 48(6):749-62. PMC: 2823977. DOI: 10.1016/j.freeradbiomed.2009.12.022. View

16.

Zillhardt M, Park S, Romero I, Sawada K, Montag A, Krausz T . Foretinib (GSK1363089), an orally available multikinase inhibitor of c-Met and VEGFR-2, blocks proliferation, induces anoikis, and impairs ovarian cancer metastasis. Clin Cancer Res. 2011; 17(12):4042-51. PMC: 3169439. DOI: 10.1158/1078-0432.CCR-10-3387. View

17.

Wang Z, Liu H, Zhang Y, Guo L, Li Z, Shi X . Anticancer potential of Euphorbia helioscopia L extracts against human cancer cells. Anat Rec (Hoboken). 2011; 295(2):223-33. DOI: 10.1002/ar.21517. View

18.

Liu G, Meng X, Jin Y, Bai J, Zhao Y, Cui X . Inhibitory role of focal adhesion kinase on anoikis in the lung cancer cell A549. Cell Biol Int. 2008; 32(6):663-70. DOI: 10.1016/j.cellbi.2008.01.292. View

19.

Li A, Ito H, Rovira I, Kim K, Takeda K, Yu Z . A role for reactive oxygen species in endothelial cell anoikis. Circ Res. 1999; 85(4):304-10. DOI: 10.1161/01.res.85.4.304. View

20.

Cusimano A, Puleio R, DAlessandro N, Loria G, McCubrey J, Montalto G . Cytotoxic activity of the novel small molecule AKT inhibitor SC66 in hepatocellular carcinoma cells. Oncotarget. 2015; 6(3):1707-22. PMC: 4359326. DOI: 10.18632/oncotarget.2738. View