Artesunate attenuates glioma proliferation, migration and invasion by affecting cellular mechanical properties

Lian,Shizhong; Shi,Ruyi; Huang,Xun; Hu,Xiaoling; Song,Bin; Bai,Yinshan; Yang,Bin; Dong,Jinyao; Du,Zhijie; Zhang,Yanyan; Jia,Junmei; Ma,Ning; Guo,Geng; Wang,Mingyu

doi:10.3892/or.2016.4847

Artesunate attenuates glioma proliferation, migration and invasion by affecting cellular mechanical properties

Authors:
- Shizhong Lian
- Ruyi Shi
- Xun Huang
- Xiaoling Hu
- Bin Song
- Yinshan Bai
- Bin Yang
- Jinyao Dong
- Zhijie Du
- Yanyan Zhang
- Junmei Jia
- Ning Ma
- Geng Guo
- Mingyu Wang
View Affiliations

Affiliations: Department of Neurosurgery, The First Hospital of Shanxi Medical University, Taiyuan, Shanxi 030001, P.R. China, Key Laboratory of Cellular Physiology, Chinese Ministry of Education, Shanxi Medical University, Taiyuan, Shanxi 030001, P.R. China, Department of Materials Science and Engineering, Jinan University, Guangzhou, Guangdong 510630, P.R. China, Basic School of Guangzhou Medical University, Guangzhou, Guangdong 511436, P.R. China, Department of General Surgery, The First Hospital of Shanxi Medical University, Taiyuan, Shanxi 030001, P.R. China, The Fourth People's Hospital of Linfen, Linfen, Shanxi 041000, P.R. China, Department of Oncology, The First Hospital of Shanxi Medical University, Taiyuan, Shanxi 030001, P.R. China
Published online on: June 2, 2016 https://doi.org/10.3892/or.2016.4847
Pages: 984-990

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

Glioma is one of the most common malignant brain tumors. Current chemotherapy is far from providing satisfactory clinical outcomes for patients with glioma. More efficient drugs are urgently needed. Artesunate (ART) is clinically used as an anti-malarial agent and exhibits potent antiproliferative activity as a traditional Chinese medicine. In addition, ART has been shown to exert a profound cytotoxic effect on various tumor cell lines, presenting a novel candidate for cancer chemotherapy. However, its anticancer effect on glioma by altering cell biomechanical properties remains unclear. The present study aimed to identify the anticancer effects of ART on human glioma SHG44 cells by assessing cell proliferation, migration/invasion, the expression of claudin-1 and the biomechanical properties of ART-treated SHG44 cells. The proliferation of the SHG44 cells was assessed by MTT assay. The cell apoptosis was detected by flow cytometry. For cell migration and invasion assays, the Transwell was used. The expression of the gene claudin-1 was detected by polymerase chain reaction. The cell membrane and biomechanical properties, as targets of ART action, were investigated by atomic force microscopy (AFM). ART significantly inhibited the proliferation of SHG44 cells in a dose- and time-dependent manner. After treatment with 30 mg/l ART, the level of cell apoptosis was significantly increased (from 6.88±0.062 to 23.7±4.16%). Furthermore, the cell migration and invasion abilities of the SHG44 cells were markedly inhibited after treatment with 30 mg/l ART. Compared with the control group (0 mg/l ART), the SHG44 cells treated with 30 mg/l ART exhibited upregulated expression of claudin-1, increased adhesive force (from 2,400±300 to 3,600±500 pN), increased high connection among SHG44 cells, increased cytomembrane roughness (from 0.118±0.011 to 0.269±0.015 µm) and reduced elasticity (from 23±8 to 3.5±1.1 MPa). The present study demonstrated that ART could alter the biomechanical properties of the glioma cells to inhibit cell proliferation, migration and invasion.

View Figures

View References

1	Louis DN: Molecular pathology of malignant gliomas. Annu Rev Pathol. 1:97–117. 2006. View Article : Google Scholar
2	Saika K and Katanoda K: Comparison of time trends in brain and central nervous system cancer mortality (1990–2006) between countries based on the WHO mortality database. Jpn J Clin Oncol. 41:304–305. 2011. View Article : Google Scholar : PubMed/NCBI
3	Sørensen MD, Fosmark S, Hellwege S, Beier D, Kristensen BW and Beier CP: Chemoresistance and chemotherapy targeting stem-like cells in malignant glioma. Adv Exp Med Biol. 853:111–138. 2015. View Article : Google Scholar : PubMed/NCBI
4	Katsetos CD, Reginato MJ, Baas PW, D'Agostino L, Legido A, Tuszyn Ski JA, Dráberová E and Dráber P: Emerging microtubule targets in glioma therapy. Semin Pediatr Neurol. 22:49–72. 2015. View Article : Google Scholar : PubMed/NCBI
5	White NJ: Qinghaosu (artemisinin): The price of success. Science. 320:330–334. 2008. View Article : Google Scholar : PubMed/NCBI
6	Youns M, Efferth T, Reichling J, Fellenberg K, Bauer A and Hoheisel JD: Gene expression profiling identifies novel key players involved in the cytotoxic effect of Artesunate on pancreatic cancer cells. Biochem Pharmacol. 78:273–283. 2009. View Article : Google Scholar : PubMed/NCBI
7	Hou J, Wang D, Zhang R and Wang H: Experimental therapy of hepatoma with artemisinin and its derivatives: In vitro and in vivo activity, chemosensitization, and mechanisms of action. Clin Cancer Res. 14:5519–5530. 2008. View Article : Google Scholar : PubMed/NCBI
8	Li LN, Zhang HD, Yuan SJ, Yang DX, Wang L and Sun ZX: Differential sensitivity of colorectal cancer cell lines to artesunate is associated with expression of beta-catenin and E-cadherin. Eur J Pharmacol. 588:1–8. 2008. View Article : Google Scholar : PubMed/NCBI
9	Nam W, Tak J, Ryu JK, Jung M, Yook JI, Kim HJ and Cha IH: Effects of artemisinin and its derivatives on growth inhibition and apoptosis of oral cancer cells. Head Neck. 29:335–340. 2007. View Article : Google Scholar
10	Liu WM, Gravett AM and Dalgleish AG: The antimalarial agent artesunate possesses anticancer properties that can be enhanced by combination strategies. Int J Cancer. 128:1471–1480. 2011. View Article : Google Scholar
11	Rinner B, Siegl V, Pürstner P, Efferth T, Brem B, Greger H and Pfragner R: Activity of novel plant extracts against medullary thyroid carcinoma cells. Anticancer Res. 24:495–500. 2004.PubMed/NCBI
12	Efferth T, Dunstan H, Sauerbrey A, Miyachi H and Chitambar CR: The anti-malarial artesunate is also active against cancer. Int J Oncol. 18:767–773. 2001.PubMed/NCBI
13	Efferth T, Sauerbrey A, Olbrich A, Gebhart E, Rauch P, Weber HO, Hengstler JG, Halatsch ME, Volm M, Tew KD, et al: Molecular modes of action of artesunate in tumor cell lines. Mol Pharmacol. 64:382–394. 2003. View Article : Google Scholar : PubMed/NCBI
14	Shi R, Cui H, Bi Y, Huang X, Song B, Cheng C, Zhang L, Liu J, He C, Wang F, et al: Artesunate altered cellular mechanical properties leading to deregulation of cell proliferation and migration in esophageal squamous cell carcinoma. Oncol Lett. 9:2249–2255. 2015.PubMed/NCBI
15	Hörber JK and Miles MJ: Scanning probe evolution in biology. Science. 302:1002–1005. 2003. View Article : Google Scholar : PubMed/NCBI
16	Alonso JL and Goldmann WH: Feeling the forces: Atomic force microscopy in cell biology. Life Sci. 72:2553–2560. 2003. View Article : Google Scholar : PubMed/NCBI
17	Lesniewska E, Milhiet PE, Giocondi MC and Le Grimellec C: Atomic force microscope imaging of cells and membranes. Methods Cell Biol. 68:51–65. 2002. View Article : Google Scholar : PubMed/NCBI
18	Soofi SS, Last JA, Liliensiek SJ, Nealey PF and Murphy CJ: The elastic modulus of Matrigel as determined by atomic force microscopy. J Struct Biol. 167:216–219. 2009. View Article : Google Scholar : PubMed/NCBI
19	Kondra S, Laishram J, Ban J, Migliorini E, Di Foggia V, Lazzarino M, Torre V and Ruaro ME: Integration of confocal and atomic force microscopy images. J Neurosci Methods. 177:94–107. 2009. View Article : Google Scholar
20	Wang J, Wan Z, Liu W, Li L, Ren L, Wang X, Sun P, Ren L, Zhao H, Tu Q, et al: Atomic force microscope study of tumor cell membranes following treatment with anti-cancer drugs. Biosens Bioelectron. 25:721–727. 2009. View Article : Google Scholar : PubMed/NCBI
21	D'Souza T, Agarwal R and Morin PJ: Phosphorylation of claudin-3 at threonine 192 by cAMP-dependent protein kinase regulates tight junction barrier function in ovarian cancer cells. J Biol Chem. 280:26233–26240. 2005. View Article : Google Scholar : PubMed/NCBI
22	Karnati HK, Panigrahi M, Shaik NA, Greig NH, Bagadi SA, Kamal MA and Kapalavayi N: Down regulated expression of claudin-1 and claudin-5 and up regulation of β-catenin: Association with human glioma progression. CNS Neurol Disord Drug Targets. 13:1413–1426. 2014. View Article : Google Scholar
23	Snedecor GW and Cochran WG: Statistical Methods. 7th edition. Iowa State University Press; Ames, IA, USA: pp. 32–43. 1980
24	Puech PH, Poole K, Knebel D and Muller DJ: A new technical approach to quantify cell-cell adhesion forces by AFM. Ultramicroscopy. 106:637–644. 2006. View Article : Google Scholar : PubMed/NCBI
25	Geiger B, Bershadsky A, Pankov R and Yamada KM: Transmembrane crosstalk between the extracellular matrix - cytoskeleton crosstalk. Nat Rev Mol Cell Biol. 2:793–805. 2001. View Article : Google Scholar : PubMed/NCBI
26	Sato K, Adachi T, Ueda D, Hojo M and Tomita Y: Measurement of local strain on cell membrane at initiation point of calcium signaling response to applied mechanical stimulus in osteoblastic cells. J Biomech. 40:1246–1255. 2007. View Article : Google Scholar
27	Voïtchovsky K, Antoranz Contera S, Kamihira M, Watts A and Ryan JF: Differential stiffness and lipid mobility in the leaflets of purple membranes. Biophys J. 90:2075–2085. 2006. View Article : Google Scholar : PubMed/NCBI
28	Fuchs E and Cleveland DW: A structural scaffolding of intermediate filaments in health and disease. Science. 279:514–519. 1998. View Article : Google Scholar : PubMed/NCBI
29	Saunders WS, Shuster M, Huang X, Gharaibeh B, Enyenihi AH, Petersen I and Gollin SM: Chromosomal instability and cytoskeletal defects in oral cancer cells. Proc Natl Acad Sci USA. 97:303–308. 2000. View Article : Google Scholar : PubMed/NCBI
30	Mahoney JA and Rosen A: Apoptosis and autoimmunity. Curr Opin Immunol. 17:583–588. 2005. View Article : Google Scholar : PubMed/NCBI
31	Gourlay CW and Ayscough KR: A role for actin in aging and apoptosis. Biochem Soc Trans. 33:1260–1264. 2005. View Article : Google Scholar : PubMed/NCBI
32	Hadj-Rabia S, Baala L, Vabres P, Hamel-Teillac D, Jacquemin E, Fabre M, Lyonnet S, De Prost Y, Munnich A, Hadchouel M, et al: Claudin-1 gene mutations in neonatal sclerosing cholangitis associated with ichthyosis: A tight junction disease. Gastroenterology. 127:1386–1390. 2004. View Article : Google Scholar : PubMed/NCBI
33	Lee JW, Hsiao WT, Chen HY, Hsu LP, Chen PR, Lin MD, Chiu SJ, Shih WL and Hsu YC: Upregulated claudin-1 expression confers resistance to cell death of nasopharyngeal carcinoma cells. Int J Cancer. 126:1353–1366. 2010.
34	Blanchard AA, Ma X, Dueck KJ, Penner C, Cooper SC, Mulhall D, Murphy LC, Leygue E and Myal Y: Claudin 1 expression in basal-like breast cancer is related to patient age. BMC Cancer. 13:2682013. View Article : Google Scholar : PubMed/NCBI
35	Kumar S and Weaver VM: Mechanics, malignancy, and metastasis: The force journey of a tumor cell. Cancer Metastasis Rev. 28:113–127. 2009. View Article : Google Scholar : PubMed/NCBI
36	Fletcher DA and Mullins RD: Cell mechanics and the cytoskeleton. Nature. 463:485–492. 2010. View Article : Google Scholar : PubMed/NCBI