Neferine treatment enhances the TRAIL‑induced apoptosis of human prostate cancer cells via autophagic flux and the JNK pathway

Nazim,Uddin  MD; Yin,Honghua; Park,Sang‑Youel

doi:10.3892/ijo.2020.5012

Neferine treatment enhances the TRAIL‑induced apoptosis of human prostate cancer cells via autophagic flux and the JNK pathway

Authors:
- Uddin MD Nazim
- Honghua Yin
- Sang‑Youel Park
View Affiliations

Affiliations: Biosafety Research Institute, College of Veterinary Medicine, Jeonbuk National University, Iksan, Jeonbuk 54596, Republic of Korea
Published online on: March 13, 2020 https://doi.org/10.3892/ijo.2020.5012
Pages: 1152-1161
Copyright: © Nazim et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Prostate cancer (PCa) is a common type of cancer among males, with a relatively high mortality rate. Tumor necrosis factor‑related apoptosis‑inducing ligand (TRAIL), a member of the tumor necrosis factor (TNF) family, initiates the apoptosis of certain cancer cells. Neferine, a primary ingredient of bisbenzylisoquinoline alkaloids, has various antitumor activities. The present study examined the effects of neferine treatment on human PCa cells. Human prostate cancer (DU145) cells were treated with neferine for 18 h, and subsequently treated with TRAIL for 2 h. Combined treatment with neferine and TRAIL significantly decreased cell viability compared to treatment with TRAIL alone. Furthermore, neferine treatment decreased the expression of p62 and increased LC3B‑II expression, as assessed by western blot analysis and immunocytochemistry. It was alsp demonstrated that neferine and TRAIL act synergistically to trigger autophagy in PCa cells, as revealed by autophagosome formation, LC3B‑II accumulation demonstrated by transmission electron microscopy (TEM) analysis and phosphorylated c‑Jun N‑terminal kinase (p‑JNK) upregulation. When the autophagic flux was attenuated by the inhibitor, chloroquine, or by genetically modified ATG5 siRNA, the enhancement of TRAIL‑induced autophagy by neferine‑induced was also attenuated. Furthermore, treatment with the JNK inhibitor, SP600125, distinctly increased the viability of the cells treated with neferine and TRAIL. On the whole, the findings of the present study demonstrate that neferine treatment effectively promotes TRAIL‑mediated cell death and this effect likely occurs via the autophagic flux and the JNK pathway.

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1	Siegel R, Naishadham D and Jemal A: Cancer statistics, 2012. CA Cancer J Clin. 62:10–29. 2012. View Article : Google Scholar : PubMed/NCBI
2	Hao L, Zhao Y, Li ZG, He HG, Liang Q, Zhang ZG, Shi ZD, Zhang PY and Han CH: Tumor necrosis factor-related apop-tosis-inducing ligand inhibits proliferation and induces apoptosis of prostate and bladder cancer cells. Oncol Lett. 13:3638–3640. 2017. View Article : Google Scholar : PubMed/NCBI
3	Nazim UM, Jeong JK and Park SY: Ophiopogonin B sensitizes TRAIL-induced apoptosis through activation of autophagy flux and downregulates cellular FLICE-like inhibitory protein. Oncotarget. 9:4161–4172. 2018. View Article : Google Scholar : PubMed/NCBI
4	Zhang S, Wang Y, Chen Z, Kim S, Iqbal S, Chi A, Ritenour C, Wang YA, Kucuk O and Wu D: Genistein enhances the efficacy of cabazitaxel chemotherapy in metastatic castration-resistant prostate cancer cells. Prostate. 73:1681–1689. 2013.PubMed/NCBI
5	Klosek M, Mertas A, Krol W, Jaworska D, Szymszal J and Szliszka E: Tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis in prostate cancer cells after treatment with xanthohumol-a natural compound present in humulus lupulus L. Int J Mol Sci. 17:pii: E837. 2016. View Article : Google Scholar : PubMed/NCBI
6	Wiley SR, Schooley K, Smolak PJ, Din WS, Huang CP, Nicholl JK, Sutherland GR, Smith TD, Rauch C, Smith CA, et al: Identification and characterization of a new member of the TNF family that induces apoptosis. Immunity. 3:673–682. 1995. View Article : Google Scholar : PubMed/NCBI
7	Wang S and El-Deiry WS: TRAIL and apoptosis induction by TNF-family death receptors. Oncogene. 22:8628–8633. 2003. View Article : Google Scholar : PubMed/NCBI
8	Kischkel FC, Lawrence DA, Chuntharapai A, Schow P, Kim KJ and Ashkenazi A: Apo2L/TRAIL-dependent recruitment of endogenous FADD and caspase-8 to death receptors 4 and 5. Immunity. 12:611–620. 2000. View Article : Google Scholar : PubMed/NCBI
9	Wei RJ, Zhang XS and He DL: Andrographolide sensitizes prostate cancer cells to TRAIL-induced apoptosis. Asian J Androl. 20:200–204. 2018. View Article : Google Scholar :
10	Sivalingam KS, Paramasivan P, Weng CF and Viswanadha VP: Neferine potentiates the antitumor effect of cisplatin in human lung adenocarcinoma cells via a mitochondria-mediated apoptosis pathway. J Cell Biochem. 118:2865–2876. 2017. View Article : Google Scholar : PubMed/NCBI
11	Poornima P, Quency RS and Padma VV: Neferine induces reactive oxygen species mediated intrinsic pathway of apoptosis in HepG2 cells. Food Chem. 136:659–667. 2013. View Article : Google Scholar
12	Poornima P, Weng CF and Padma VV: Neferine, an alkaloid from lotus seed embryo, inhibits human lung cancer cell growth by MAPK activation and cell cycle arrest. Biofactors. 40:121–131. 2014. View Article : Google Scholar
13	Zhang X, Liu Z, Xu B, Sun Z, Gong Y and Shao C: Neferine, an alkaloid ingredient in lotus seed embryo, inhibits proliferation of human osteosarcoma cells by promoting p38 MAPK-mediated p21 stabilization. Eur J Pharmacol. 677:47–54. 2012. View Article : Google Scholar : PubMed/NCBI
14	Li T, Su L, Zhong N, Hao X, Zhong D, Singhal S and Liu X: Salinomycin induces cell death with autophagy through activation of endoplasmic reticulum stress in human cancer cells. Autophagy. 9:1057–1068. 2013. View Article : Google Scholar : PubMed/NCBI
15	Ouyang L, Shi Z, Zhao S, Wang FT, Zhou TT, Liu B and Bao JK: Programmed cell death pathways in cancer: A review of apoptosis, autophagy and programmed necrosis. Cell Prolif. 45:487–498. 2012. View Article : Google Scholar : PubMed/NCBI
16	Liu YP, Li L, Xu L, Dai EN and Chen WD: Cantharidin suppresses cell growth and migration, and activates autophagy in human non-small cell lung cancer cells. Oncol Lett. 15:6527–6532. 2018.PubMed/NCBI
17	White E: The role for autophagy in cancer. J Clin Invest. 125:42–46. 2015. View Article : Google Scholar : PubMed/NCBI
18	Parzych KR and Klionsky DJ: An overview of autophagy: morphology, mechanism, and regulation. Antioxid Redox Signal. 20:460–473. 2014. View Article : Google Scholar :
19	Fan S, Qi M, Yu Y, Li L, Yao G, Tashiro S, Onodera S and Ikejima T: P53 activation plays a crucial role in silibinin induced ROS generation via PUMA and JNK. Free Radic Res. 46:310–319. 2012. View Article : Google Scholar : PubMed/NCBI
20	Kuntz S, Wenzel U and Daniel H: Comparative analysis of the effects of flavonoids on proliferation, cytotoxicity, and apoptosis in human colon cancer cell lines. Eur J Nutr. 38:133–142. 1999. View Article : Google Scholar : PubMed/NCBI
21	Bonavida B, Ng C, Jazirehi A, Schiller G and Mizutani Y: Selectivity of TRAIL-mediated apoptosis of cancer cells and synergy with drugs: The trail to non-toxic cancer therapeutics. Int J Oncol. 15:793–1595. 1999.PubMed/NCBI
22	Ji D, Zhang Z, Cheng L, Chang J, Wang S, Zheng B, Zheng R, Sun Z, Wang C, Zhang Z, et al: The combination of RAD001 and MK-2206 exerts synergistic cytotoxic effects against PTEN mutant gastric cancer cells: Involvement of MAPK-dependent autophagic, but not apoptotic cell death pathway. PLoS One. 9:e851162014. View Article : Google Scholar : PubMed/NCBI
23	Nazim UM, Moon JH, Lee JH, Lee YJ, Seol JW, Eo SK, Lee JH and Park SY: Activation of autophagy flux by metformin down-regulates cellular FLICE-like inhibitory protein and enhances TRAIL-induced apoptosis. Oncotarget. 7:23468–23481. 2016. View Article : Google Scholar : PubMed/NCBI
24	Labsch S, Liu L, Bauer N, Zhang Y, Aleksandrowicz E, Gladkich J, Schönsiegel F and Herr I: Sulforaphane and TRAIL induce a synergistic elimination of advanced prostate cancer stem-like cells. Int J Oncol. 44:1470–1480. 2014. View Article : Google Scholar : PubMed/NCBI
25	Lagadec C, Adriaenssens E, Toillon R, Chopin V, Romon R, Van Coppenolle F, Hondermarck H and Le Bourhis X: Tamoxifen and TRAIL synergistically induce apoptosis in breast cancer cells. Oncogene. 27:1472–1477. 2008. View Article : Google Scholar
26	Zhu H, Ding WJ, Wu R, Weng QJ, Lou JS, Jin RJ, Lu W, Yang B and He QJ: Synergistic anti-cancer activity by the combination of TRAIL/APO-2L and celastrol. Cancer Invest. 28:23–32. 2010. View Article : Google Scholar
27	Oh YT, Yue P, Wang D, Tong JS, Chen ZG, Khuri FR and Sun SY: Suppression of death receptor 5 enhances cancer cell invasion and metastasis through activation of caspase-8/TRAF2-mediated signaling. Oncotarget. 6:41324–41338. 2015. View Article : Google Scholar : PubMed/NCBI
28	Han B, Yao W, Oh YT, Tong JS, Li S, Deng J, Yue P, Khuri FR and Sun SY: The novel proteasome inhibitor carfilzomib activates and enhances extrinsic apoptosis involving stabilization of death receptor 5. Oncotarget. 6:17532–17542. 2015. View Article : Google Scholar : PubMed/NCBI
29	von Karstedt S, Montinaro A and Walczak H: Exploring the TRAILs less travelled: TRAIL in cancer biology and therapy. Nature reviews. Cancer. 17:352–366. 2017.
30	Selvarajoo K: A systems biology approach to overcome TRAIL resistance in cancer treatment. Prog Biophys Mol Biol. 128:142–154. 2017. View Article : Google Scholar : PubMed/NCBI
31	Poornima P, Weng CF and Padma VV: Neferine from Nelumbo nucifera induces autophagy through the inhibition of PI3K/Akt/mTOR pathway and ROS hyper generation in A549 cells. Food Chem. 141:3598–3605. 2013. View Article : Google Scholar : PubMed/NCBI
32	Klionsky DJ and Emr SD: Autophagy as a regulated pathway of cellular degradation. Science. 290:1717–1721. 2000. View Article : Google Scholar : PubMed/NCBI
33	Petersen A, Larsen KE, Behr GG, Romero N, Przedborski S, Brundin P and Sulzer D: Expanded CAG repeats in exon 1 of the Huntington's disease gene stimulate dopamine-mediated striatal neuron autophagy and degeneration. Hum Mol Genet. 10:1243–1254. 2001. View Article : Google Scholar : PubMed/NCBI
34	Kim SW, Lee JH, Moon JH, Nazim UM, Lee YJ, Seol JW, Hur J, Eo SK, Lee JH and Park SY: Niacin alleviates TRAIL-mediated colon cancer cell death via autophagy flux activation. Oncotarget. 7:4356–4368. 2016.
35	Zhang Y, Wu Y, Cheng Y, Zhao Z, Tashiro S, Onodera S and Ikejima T: Fas-mediated autophagy requires JNK activation in HeLa cells. Biochem Biophys Res Commun. 377:1205–1210. 2008. View Article : Google Scholar : PubMed/NCBI
36	Stuckey DW and Shah K: TRAIL on trial: Preclinical advances in cancer therapy. Trends Mol Med. 19:685–694. 2013. View Article : Google Scholar : PubMed/NCBI
37	Xu L, Zhang X, Li Y, Lu S, Lu S, Li J, Wang Y, Tian X, Wei JJ, Shao C and Liu Z: Neferine induces autophagy of human ovarian cancer cells via p38 MAPK/JNK activation. Tumour Biol. 37:8721–8729. 2016. View Article : Google Scholar : PubMed/NCBI
38	Musumeci G, Castrogiovanni P, Trovato FM, Weinberg AM, Al-Wasiyah MK, Alqahtani MH and Mobasheri A: Biomarkers of chondrocyte apoptosis and autophagy in osteoarthritis. Int J Mol Sci. 16:20560–20575. 2015. View Article : Google Scholar : PubMed/NCBI
39	Fiandalo MV and Kyprianou N: Caspase control: Protagonists of cancer cell apoptosis. Exp Oncol. 34:165–175. 2012.PubMed/NCBI
40	Huang K: Mechanism of Bax/Bak Activation in Apoptotic Signaling (unpublished PhD thesis). University of Nebraska Medical Center; 2019
41	Ly JD, Grubb DR and Lawen A: The mitochondrial membrane potential (deltapsi(m)) in apoptosis; an update. Apoptosis. 8:115–128. 2003. View Article : Google Scholar : PubMed/NCBI
42	Tounekti O, Belehradek J Jr and Mir L: Relationships between DNA fragmentation, chromatin condensation, and changes in flow cytometry profiles detected during apoptosis. Exp Cell Res. 217:506–516. 1995. View Article : Google Scholar : PubMed/NCBI
43	Yoshii SR and Mizushima N: Monitoring and measuring autophagy. Int J Mol Sci. 18:18652017. View Article : Google Scholar :
44	Tanida I, Ueno T and Kominami E: LC3 and autophagy. Methods Mol Biol. 445:77–88. 2008. View Article : Google Scholar : PubMed/NCBI
45	Ganesher A, Chaturvedi P, Sahai R, Meena S, Mitra K, Datta D and Panda G: New spisulosine derivative promotes robust autophagic response to cancer cells. Eur J Med Chem. 188:1120112020. View Article : Google Scholar : PubMed/NCBI
46	Han X, Guo L, Jiang X, Wang Y, Wang Z and Li D: Curcumin inhibits cell viability by inducing apoptosis and autophagy in human colon cancer cells. Proceed Anticancer Res. 3:21–25. 2019.
47	Chiou JT, Huang CH, Lee YC, Wang LJ, Shi YJ, Chen YJ and Chang LS: Compound C induces autophagy and apoptosis in parental and hydroquinone-selected malignant leukemia cells through the ROS/p38 MAPK/AMPK/TET2/FOXP3 axis. Cell Biol Toxicol. Jan 3–2020.Epub ahead of prin. View Article : Google Scholar : PubMed/NCBI
48	Opipari AW Jr, Tan L, Boitano AE, Sorenson DR, Aurora A and Liu JR: Resveratrol-induced autophagocytosis in ovarian cancer cells. Cancer Res. 64:696–703. 2004. View Article : Google Scholar : PubMed/NCBI