Hexane fraction of Pluchea indica root extract inhibits proliferation and induces autophagy in human glioblastoma cells

Cho,Chung-Lung; Lee,Ya-Zhe; Tseng,Chao-Neng; Cho,Joshua; Cheng,Yuan-Bin; Wang,Kuo‑Wei; Chen,Han-Jung; Chiou,Shean-Jaw; Chou,Chia-Hua; Hong,Yi-Ren

doi:10.3892/br.2017.979

Hexane fraction of Pluchea indica root extract inhibits proliferation and induces autophagy in human glioblastoma cells

Authors:
- Chung-Lung Cho
- Ya-Zhe Lee
- Chao-Neng Tseng
- Joshua Cho
- Yuan-Bin Cheng
- Kuo‑Wei Wang
- Han-Jung Chen
- Shean-Jaw Chiou
- Chia-Hua Chou
- Yi-Ren Hong
View Affiliations

Affiliations: Department of Biological Sciences, National Sun Yat-sen University, Kaohsiung 80424, Taiwan, R.O.C., University of Pennsylvania School of Dental Medicine, Philadelphia, PA 19104, USA, Graduate Institute of Natural Products, School of Pharmacy, Kaohsiung Medical University, Kaohsiung 80708, Taiwan, R.O.C., Department of Neurosurgery, E-Da Hospital, I-Shou University, Kaohsiung 84001, Taiwan, R.O.C., Department of Biochemistry, College of Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan, R.O.C., Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan, R.O.C.
Published online on: September 11, 2017 https://doi.org/10.3892/br.2017.979
Pages: 416-422

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

Pluchea indica (L.) Less. is a perennial plant known for its versatile uses in traditional medicine. Previous findings have shown that the extracts of Pluchea indica possess significant anti-inflammatory, anti-ulcer and anti-tuberculosis activity. The aim of this study was to demonstrate the anticancer activity of the hexane fraction of P. indica root extract (H-PIRE) in human glioblastoma cells using flow cytometric and western blot analysis. The results shoewd that, H-PIRE suppressed the growth of glioblastoma cells in a dose-dependent manner. H-PIRE treatment markedly decreased the population of cells in S and G2/M phases. The significant upregulation of acidic vesicular organelle (AVO) was detected during H-PIRE treatment. The expression levels of microtubule-associated light chain 3-II (LC3-II) protein, phosphorylated JNK and phosphorylated p38 were significantly increased, confirming the occurrence of autophagy during the process. Finally, the combination treatment of H-PIRE and LY294002, a pan PI3K inhibitor, further decreased cell viability, suggesting an additive anticancer effect. Taken together, our results suggest that H-PIRE suppresses the proliferation of glioblastoma cells by inducing cell cycle arrest and autophagy.

View Figures

View References

1	Bleeker FE, Molenaar RJ and Leenstra S: Recent advances in the molecular understanding of glioblastoma. J Neurooncol. 108:11–27. 2012. View Article : Google Scholar : PubMed/NCBI
2	Bleeker FE, Lamba S, Zanon C, Molenaar RJ, Hulsebos TJM, Troost D, van Tilborg AA, Vandertop WP, Leenstra S, van Noorden CJF, et al: Mutational profiling of kinases in glioblastoma. BMC Cancer. 14:7182014. View Article : Google Scholar : PubMed/NCBI
3	Crawford JR, Santi MR, Thorarinsdottir HK, Cornelison R, Rushing EJ, Zhang H, Yao K, Jacobson S and Macdonald TJ: Detection of human herpesvirus-6 variants in pediatric brain tumors: Association of viral antigen in low grade gliomas. J Clin Virol. 46:37–42. 2009. View Article : Google Scholar : PubMed/NCBI
4	Baglietto L, Giles GG, English DR, Karahalios A, Hopper JL and Severi G: Alcohol consumption and risk of glioblastoma; evidence from the Melbourne Collaborative Cohort Study. Int J Cancer. 128:1929–1934. 2011. View Article : Google Scholar : PubMed/NCBI
5	Braganza MZ, Kitahara CM, de González Berrington A, Inskip PD, Johnson KJ and Rajaraman P: Ionizing radiation and the risk of brain and central nervous system tumors: A systematic review. Neuro-oncol. 14:1316–1324. 2012. View Article : Google Scholar : PubMed/NCBI
6	Ellis HP, Greenslade M, Powell B, Spiteri I, Sottoriva A and Kurian KM: Current challenges in glioblastoma: Intratumour heterogeneity, residual disease and models to predict disease recurrence. Front Oncol. 5:2512015. View Article : Google Scholar : PubMed/NCBI
7	Johnson DR and O'Neill BP: Glioblastoma survival in the United States before and during the temozolomide era. J Neurooncol. 107:359–364. 2012. View Article : Google Scholar : PubMed/NCBI
8	Wrensch M, Minn Y, Chew T, Bondy M and Berger MS: Epidemiology of primary brain tumors: Current concepts and review of the literature. Neuro Oncol. 4:278–299. 2002. View Article : Google Scholar : PubMed/NCBI
9	Emsen B, Aslan A, Togar B and Turkez H: In vitro antitumor activities of the lichen compounds olivetoric, physodic and psoromic acid in rat neuron and glioblastoma cells. Pharm Biol. 54:1748–1762. 2016. View Article : Google Scholar : PubMed/NCBI
10	Mulpur BH, Nabors LB, Thompson RC, Olson JJ, LaRocca RV, Thompson Z and Egan KM: Complementary therapy and survival in glioblastoma. Neurooncol Pract. 2:122–126. 2015.PubMed/NCBI
11	Sen T, Dhara AK, Bhattacharjee S, Pal S and Chaudhuri Nag AK: Antioxidant activity of the methanol fraction of Pluchea indica root extract. Phytother Res. 16:331–335. 2002. View Article : Google Scholar : PubMed/NCBI
12	Rahman Ab MR, Razak Abdul F and Bakri Mohd M: Evaluation of wound closure activity of Nigella sativa, Melastoma malabathricum, Pluchea indica and Piper sarmentosum extracts on scratched monolayer of human gingival fibroblasts. Evid Based Complement Alternat Med. 2014:1903422014. View Article : Google Scholar : PubMed/NCBI
13	Arsiningtyas IS, Gunawan-Puteri MD, Kato E and Kawabata J: Identification of α-glucosidase inhibitors from the leaves of Pluchea indica (L.) Less., a traditional Indonesian herb: Promotion of natural product use. Nat Prod Res. 28:1350–1353. 2014. View Article : Google Scholar : PubMed/NCBI
14	Gomes A, Saha A, Chatterjee I and Chakravarty AK: Viper and cobra venom neutralization by β-sitosterol and stigmasterol isolated from the root extract of Pluchea indica Less. (Asteraceae). Phytomedicine. 14:637–643. 2007. View Article : Google Scholar : PubMed/NCBI
15	Biswas R, Dutta PK, Achari B, Bandyopadhyay D, Mishra M, Pramanik KC and Chatterjee TK: Isolation of pure compound R/J/3 from Pluchea indica (L.) Less. and its anti-amoebic activities against Entamoeba histolytica. Phytomedicine. 14:534–537. 2007. View Article : Google Scholar : PubMed/NCBI
16	Sen T, Ghosh TK and Chaudhuri AK: Studies on the mechanism of anti-inflammatory and anti-ulcer activity of Pluchea indica-probable involvement of 5-lipooxygenase pathway. Life Sci. 52:737–743. 1993. View Article : Google Scholar : PubMed/NCBI
17	Mohamad S, Zin NM, Wahab HA, Ibrahim P, Sulaiman SF, Zahariluddin ASM and Noor SSM: Antituberculosis potential of some ethnobotanically selected Malaysian plants. J Ethnopharmacol. 133:1021–1026. 2011. View Article : Google Scholar : PubMed/NCBI
18	Locher CP, Witvrouw M, De Béthune MP, Burch MT, Mower HF, Davis H, Lasure A, Pauwels R, De Clercq E and Vlietinck AJ: Antiviral activity of Hawaiian medicinal plants against human immunodeficiency Virus Type-1 (HIV-1). Phytomedicine. 2:259–264. 1996. View Article : Google Scholar : PubMed/NCBI
19	Ohtsuki T, Yokosawa E, Koyano T, Preeprame S, Kowithayakorn T, Sakai S, Toida T and Ishibashi M: Quinic acid esters from Pluchea indica with collagenase, MMP-2 and −9 inhibitory activities. Phytother Res. 22:264–266. 2008. View Article : Google Scholar : PubMed/NCBI
20	Cho JJ, Cho CL, Kao CL, Chen CM, Tseng CN, Lee YZ, Liao LJ and Hong YR: Crude aqueous extracts of Pluchea indica (L.) Less. inhibit proliferation and migration of cancer cells through induction of p53-dependent cell death. BMC Complement Altern Med. 12:2652012. View Article : Google Scholar : PubMed/NCBI
21	Kao CL, Cho J, Lee YZ, Cheng YB, Chien CY, Hwang CF, Hong YR, Tseng CN and Cho CL: Ethanolic extracts of Pluchea indica induce apoptosis and antiproliferation effects in human nasopharyngeal carcinoma cells. Molecules. 20:11508–11523. 2015. View Article : Google Scholar : PubMed/NCBI
22	Harborne JB: Phytochemical methods: a guide to modern techniques of plant analysis. 3rd. Chapman and Hall; London, UK: 1998, View Article : Google Scholar
23	Butler LG, Price ML and Brotherton JE: Vanillin assay for proanthocyanidins (condensed tannins): Modification of the solvent for estimation of the degree of polymerization. J Agric Food Chem. 30:1087–1089. 1982. View Article : Google Scholar
24	Pavon LF, Marti LC, Sibov TT, Malheiros SM, Brandt RA, Cavalheiro S and Gamarra LF: In vitro analysis of neurospheres derived from gioblastoma primary culture: A novel methodology paradigm. Front Neurol. 4:2142014. View Article : Google Scholar : PubMed/NCBI
25	Wink M, Ashour ML and El-Readi MZ: Secondary metabolites from plants inhibiting ABC transporters and reversing resistance of cancer cells and microbes to cytotoxic and antimicrobial agents. Front Microbiol. 3:1302012. View Article : Google Scholar : PubMed/NCBI
26	Tabrez S, Priyadarshini M, Urooj M, Shakil S, Ashraf GM, Khan MS, Kamal MA, Alam Q, Jabir NR, Abuzenadah AM, et al: Cancer chemoprevention by polyphenols and their potential application as nanomedicine. J Environ Sci Health C Environ Carcinog Ecotoxicol Rev. 31:67–98. 2013. View Article : Google Scholar : PubMed/NCBI
27	Hansen TE and Johansen T: Following autophagy step by step. BMC Biol. 9:392011. View Article : Google Scholar : PubMed/NCBI
28	Takeuchi H, Kondo Y, Fujiwara K, Kanzawa T, Aoki H, Mills GB and Kondo S: Synergistic augmentation of rapamycin-induced autophagy in malignant glioma cells by phosphatidylinositol 3-kinase/protein kinase B inhibitors. Cancer Res. 65:3336–3346. 2005. View Article : Google Scholar : PubMed/NCBI
29	Berger Z, Ravikumar B, Menzies FM, Oroz LG, Underwood BR, Pangalos MN, Schmitt I, Wullner U, Evert BO, O'Kane CJ, et al: Rapamycin alleviates toxicity of different aggregate-prone proteins. Hum Mol Genet. 15:433–442. 2006. View Article : Google Scholar : PubMed/NCBI
30	Cuadrado A and Nebreda AR: Mechanisms and functions of p38 MAPK signalling. Biochem J. 429:403–417. 2010. View Article : Google Scholar : PubMed/NCBI
31	Sun T, Li D, Wang L, Xia L, Ma J, Guan Z, Feng G and Zhu X: c-Jun NH2-terminal kinase activation is essential for up-regulation of LC3 during ceramide-induced autophagy in human nasopharyngeal carcinoma cells. J Transl Med. 9:1612011. View Article : Google Scholar : PubMed/NCBI
32	Zhou C, Zhou J, Sheng F, Zhu H, Deng X, Xia B and Lin J: The heme oxygenase-1 inhibitor ZnPPIX induces non-canonical, Beclin 1-independent, autophagy through p38 MAPK pathway. Acta Biochim Biophys Sin (Shanghai). 44:815–822. 2012. View Article : Google Scholar : PubMed/NCBI
33	Sui X, Kong N, Ye L, Han W, Zhou J, Zhang Q, He C and Pan H: p38 and JNK MAPK pathways control the balance of apoptosis and autophagy in response to chemotherapeutic agents. Cancer Lett. 344:174–179. 2014. View Article : Google Scholar : PubMed/NCBI
34	Tanida I: Autophagosome formation and molecular mechanism of autophagy. Antioxid Redox Signal. 14:2201–2214. 2011. View Article : Google Scholar : PubMed/NCBI
35	Shimizu S, Yoshida T, Tsujioka M and Arakawa S: Autophagic cell death and cancer. Int J Mol Sci. 15:3145–3153. 2014. View Article : Google Scholar : PubMed/NCBI
36	Koukourakis MI, Mitrakas AG and Giatromanolaki A: Therapeutic interactions of autophagy with radiation and temozolomide in glioblastoma: Evidence and issues to resolve. Br J Cancer. 114:485–496. 2016. View Article : Google Scholar : PubMed/NCBI
37	Yan Y, Xu Z, Dai S, Qian L, Sun L and Gong Z: Targeting autophagy to sensitive glioma to temozolomide treatment. J Exp Clin Cancer Res. 35:232016. View Article : Google Scholar : PubMed/NCBI
38	Nicholson KM and Anderson NG: The protein kinase B/Akt signalling pathway in human malignancy. Cell Signal. 14:381–395. 2002. View Article : Google Scholar : PubMed/NCBI
39	Morgensztern D and McLeod HL: PI3K/Akt/mTOR pathway as a target for cancer therapy. Anticancer Drugs. 16:797–803. 2005. View Article : Google Scholar : PubMed/NCBI