Effects of ginkgol C17:1 on cisplatin‑induced autophagy and apoptosis in HepG2 cells

Liu,Jun; Li,Yueying; Yang,Xiaoming; Dong,Yan; Wu,Jing; Chen,Min

doi:10.3892/ol.2017.7398

Effects of ginkgol C17:1 on cisplatin‑induced autophagy and apoptosis in HepG2 cells

Authors:
- Jun Liu
- Yueying Li
- Xiaoming Yang
- Yan Dong
- Jing Wu
- Min Chen
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Affiliations: Department of Biology, School of Medicine, Jiangsu University, Zhenjiang, Jiangsu 212013, P.R. China, Department of Food Science and Engineering, School of Food and Biological Engineering, Jiangsu University, Zhenjiang, Jiangsu 212013, P.R. China, Department of Physiology, School of Medicine, Jiangsu University, Zhenjiang, Jiangsu 212013, P.R. China, Department of Medical Technology, Jingjiang College, Jiangsu University, Zhenjiang, Jiangsu 212013, P.R. China
Published online on: November 14, 2017 https://doi.org/10.3892/ol.2017.7398
Pages: 1021-1029
Copyright: © Liu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Previous studies have demonstrated that ginkgol C17:1 significantly inhibits human liver cancer cells and enhances the anticancer activity of cisplatin in vivo and in vitro. However, the mechanism and biological function of ginkgol C17:1 on cells undergoing chemotherapy remain unclear. The aim of the present study was to determine the antitumor activity and mechanism of ginkgol C17:1 in combination with cisplatin in human hepatoblastoma HepG2 cells. The green fluorescent protein (GFP)‑light chain 3 (LC3) adenovirus was transfected into HepG2 cells and autophagic flux was determined using fluorescence microscopy. Western blot analysis was also conducted to measure the expression of proteins associated with apoptosis, autophagy and their associated signaling pathways. Compared with the control group, autophagic flux and nucleus aberration rates were significantly increased (P<0.05), and the expression of proteins associated with autophagy and apoptosis were increased in the groups treated with cisplatin or ginkgol C17:1, respectively. However, following co‑treatment with ginkgol C17:1 and cisplatin, the autophagic flux and the expression of autophagy proteins decreased; however, the nucleus aberration rate and apoptosis protein expression significantly increased (P<0.05) compared with the group treated with cisplatin alone. Additionally, the signaling pathways of autophagy and apoptosis were also activated following treatment with cisplatin, alone and in combination with ginkgol C17:1. Taken together, these results indicate that ginkgol C17:1 inhibits cisplatin‑induced autophagy via AMP‑activated protein kinase/ULK1signaling and increases cisplatin‑induced apoptosis in HepG2 cells via the phosphoinositide 3‑kinase/Akt/mechanistic target of rapamycin pathway.

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1	Farazi PA and Depinho RA: Hepatocellular carcinoma pathogenesis: From genes to environment. Nat Rev Cancer. 6:674–687. 2006. View Article : Google Scholar : PubMed/NCBI
2	Khaderi S, Guiteau J, Cotton RT, O'Mahony C, Rana A and Goss JA: Role of liver transplantation in the management of hepatoblastoma in the pediatric population. World J Transplant. 4:294–298. 2014. View Article : Google Scholar : PubMed/NCBI
3	Abdul HA, Lai ML and Phaik LC: Tissue microarray immunohistochemical profiles of p53 and pRB in hepatocellular carcinoma and hepatoblastoma. Asian Pac J Cancer Prev. 15:3959–3963. 2014. View Article : Google Scholar : PubMed/NCBI
4	López-Terrada D, Cheung SW, Finegold MJ and Knowles BB: Hep G2 is a hepatoblastoma-derived cell line. Hum Pathol. 40:1512–1517. 2009. View Article : Google Scholar
5	Pang RTK, Poon TCW, Wong N, Lai PSB, Wong NLY, Chan CML, Yu JWS, Chan ATC and Sung JJY: Comparison of protein expression patterns between hepatocellular carcinoma cell lines and a hepatoblastoma cell line. Clin Proteomics. 1:313–331. 2004. View Article : Google Scholar
6	Rishi RR, Kimberlee KS, Ruth IH, Michael SB, Max B and Lisa EH: Hepatoblastoma: A need for cell lines and tissue banks to develop targeted drug therapies. Front Pediatr. 4:222016.PubMed/NCBI
7	Pateva IB, Egler RA and Stearns DS: Hepatoblastoma in an 11-year-old: Case report and a review of the literature. Medicine (Baltimore). 96:e58582017. View Article : Google Scholar : PubMed/NCBI
8	Tomlinson GE and Kappler R: Genetics and epigenetics of hepatoblastoma. Pediatr Blood Cancer. 59:785–792. 2012. View Article : Google Scholar : PubMed/NCBI
9	Boulikas T and Vougiouka M: Cisplatin and platinum drugs at the molecular level (Review). Oncol Rep. 10:1663–1682. 2003.PubMed/NCBI
10	Amable L: Cisplatin resistance and opportunities for precision medicine. Pharmacol Res. 106:27–36. 2016. View Article : Google Scholar : PubMed/NCBI
11	Muggia F: Platinum compounds 30 years after the introduction of cisplatin: Implications for the treatment of ovarian cancer. Gynecol Oncol. 112:275–281. 2009. View Article : Google Scholar : PubMed/NCBI
12	Zhang HQ, Fang N, Liu XM, Xiong SP, Liao YQ, Jin WJ, Song RF and Wan YY: Antitumor activity of chloroquine in combination with Cisplatin in human gastric cancer xenografts. Asian Pac J Cancer Prev. 16:3907–3912. 2015. View Article : Google Scholar : PubMed/NCBI
13	García-Cano J, Ambroise G, Pascual-Serra R, Carrión MC, Serrano-Oviedo L, Ortega-Muelas M, Cimas FJ, Sabater S, Ruiz-Hidalgo MJ, Sanchez Perez I, et al: Exploiting the potential of autophagy in cisplatin therapy: A new strategy to overcome resistance. Oncotarget. 6:15551–15565. 2015. View Article : Google Scholar : PubMed/NCBI
14	Lu SZ and Harrison-Findik DD: Autophagy and cancer. World J Biol Chem. 4:64–70. 2013. View Article : Google Scholar : PubMed/NCBI
15	Mizushima N and Komatsu M: Autophagy: Renovation of cells and tissues. Cell. 147:728–741. 2011. View Article : Google Scholar : PubMed/NCBI
16	Mathew R, Karp CM, Beaudoin B, Vuong N, Chen G, Chen HY, Bray K, Reddy A, Bhanot G, Gelinas C, et al: Autophagy suppresses tumorigenesis through elimination of p62. Cell. 137:1062–1075. 2009. View Article : Google Scholar : PubMed/NCBI
17	Liu M, Ma S, Liu M, Hou Y, Liang B, Su X and Liu X: Synergistic killing of lung cancer cells by cisplatin and radiation via autophagy and apoptosis. Oncol Lett. 7:1903–1910. 2014.PubMed/NCBI
18	Sakamoto A and Iwamoto Y: Current status and perspectives regarding the treatment of osteo-sarcoma: Chemotherapy. Rev Recent Clin Trials. 3:228–231. 2008. View Article : Google Scholar : PubMed/NCBI
19	Choi AM, Ryter SW and Levine B: Autophagy in human health and disease. N Engl J Med. 368:651–662. 2013. View Article : Google Scholar : PubMed/NCBI
20	Chen J, Wang Q, Yin FQ, Zhang W, Yan LH and Li L: MTRR silencing inhibits growth and cisplatin resistance of ovarian carcinoma via inducing apoptosis and reducing autophagy. Am J Transl Res. 7:1510–1527. 2015.PubMed/NCBI
21	Vanhaesebroeck B, Stephens L and Hawkins P: PI3K signalling: The path to discovery and understanding. Nat Rev Mol Cell Biol. 13:195–203. 2012. View Article : Google Scholar : PubMed/NCBI
22	Tsai JP, Lee CH, Ying TH, Lin CL, Lin CL, Hsueh JT and Hsieh YH: Licochalcone A induces autophagy through PI3K/Akt/mTOR inactivation and autophagy suppression enhances Licochalcone A-induced apoptosis of human cervical cancer cells. Oncotarget. 6:28851–28866. 2015. View Article : Google Scholar : PubMed/NCBI
23	Laplante M and Sabatini DM: mTOR signaling in growth control and disease. Cell. 149:274–293. 2012. View Article : Google Scholar : PubMed/NCBI
24	Nash KM and Shah ZA: Current perspectives on the beneficial role of ginkgo biloba in neurological and cerebrovascular disorders. Integr Med InsIghts. 10:1–9. 2015. View Article : Google Scholar : PubMed/NCBI
25	Chen Q, Yang GW and An LG: Apoptosis of hepatoma cells SMMC-7721 induced by ginkgo biloba seed polysaccharide. World J Gastroenterol. 8:832–836. 2002. View Article : Google Scholar : PubMed/NCBI
26	Yang XM, Wang YF, Li YY and Ma HL: Thermal stability of ginkgolic acids from ginkgo biloba and the effects of ginkgol C17:1 on the apoptosis and migration of SMMMC7721 cells. Fitoterapia. 98:66–76. 2014. View Article : Google Scholar : PubMed/NCBI
27	Ma X, Liu H, Foyi SR, Godar RJ, Weinheimer CJ, Hill JA and Diwan A: Impaired autophagosome clearance contributes to cardiomyocyte death in ischemia/reperfusion injury. Circulation. 125:3170–3181. 2012. View Article : Google Scholar : PubMed/NCBI
28	Reggiori F, Komatsu M, Finley K and Simonsen A: Autophagy: More than a nonselective pathway. Int J Cell Biol. 2012:2196252012. View Article : Google Scholar : PubMed/NCBI
29	Kabeya Y, Mizushima N, Ueno T, Yamamoto A, Kirisako T, Noda T, Kominami E, Ohsumi Y and Yoshimori T: LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. EMBO J. 19:5720–5728. 2000. View Article : Google Scholar : PubMed/NCBI
30	Lindqvist LM, Simon AK and Baehrecke EH: Current questions and possible controversies in autophagy. Cell Death Discov. 1:pii: 150362015. View Article : Google Scholar
31	Zhang Z, Shao Z, Xiong L and Yang S: Inhibition of autophagy enhances cisplatin-induced apoptosis in the MG63 human osteosarcoma cell line. Oncol Lett. 10:2941–2946. 2015.PubMed/NCBI
32	Amaravadi RK and Thompson CB: The roles of therapy-induced autophagy and necrosis in cancer treatment. Clin Cancer Res. 13:7271–7279. 2007. View Article : Google Scholar : PubMed/NCBI
33	Bao L, Jaramillo MC, Zhang Z, Zheng Y, Yao M, Zhang DD and Yi X: Induction of autophagy contributes to cisplatin resistance in human ovarian cancer cells. Mol Med Rep. 11:91–98. 2015. View Article : Google Scholar : PubMed/NCBI
34	Le Grazie M, Biagini MR, Tarocchi M, Polvani S and Galli A: Chemotherapy for hepatocellular carcinoma: The present and the future. World J Hepatol. 9:907–920. 2017. View Article : Google Scholar : PubMed/NCBI
35	Cavallo F, Feldman DR and Barchi M: Revisiting DNA damage repair, p53-mediated apoptosis and cisplatin sensitivity in germ cell tumors. Int J Dev Biol. 57:273–280. 2013. View Article : Google Scholar : PubMed/NCBI
36	Zhang HQ, He B, Fang N, Lu S, Liao YQ and Wan YY: Autophagy inhibition sensitizes cisplatin cytotoxicity in human gastric cancer cell line SGC7901. Asian Pac J Cancer Prev. 14:4685–4688. 2013. View Article : Google Scholar : PubMed/NCBI
37	Kimura T, Takabatake Y, Takahashi A and Isaka Y: Chloroquine in cancer therapy: A double-edged sword of autophagy. Cancer Res. 73:3–7. 2013. View Article : Google Scholar : PubMed/NCBI
38	Li YY, Liu J, Yang XM, Dong Y, Liu YL and Chen M: Ginkgol C17:1 inhibits tumor growth by blunting the EGF-PI3K/Akt signaling pathway. J Biomed Res. 31:232–239. 2017.PubMed/NCBI
39	Tang JY, Dai T, Zhang H, Xiong WJ, Xu MZ, Wang XJ, Tang QH, Chen B and Xu M: GDC-0980-induced apoptosis is enhanced by autophagy inhibition in human pancreatic cancer cells. Biochem Biophys Res Commun. 453:533–538. 2014. View Article : Google Scholar : PubMed/NCBI
40	Wu HM, Jiang ZF, Ding PS, Shao LJ and Liu RY: Hypoxia-induced autophagy mediates cisplatin resistance in lung cancer cells. Sci Rep. 5:122912015. View Article : Google Scholar : PubMed/NCBI
41	Yan MM, Ni JD, Song D, Ding M and Huang J: Interplay between unfolded protein response and autophagy promotes tumor drug resistance. Oncol Lett. 10:1959–1969. 2015.PubMed/NCBI
42	Luo J, Manning BD and Cantley LC: Targeting the PI3K-Akt pathway in human cancer: Rationale and promise. Cancer Cell. 4:257–262. 2003. View Article : Google Scholar : PubMed/NCBI
43	Li F, Zeng J, Gao Y, Guan Z, Ma Z, Shi Q, Du C, Jia J, Xu S, Wang X, et al: G9a inhibition induces autophagic cell death via AMPK/mTOR pathway in bladder transitional cell carcinoma. PLoS One. 10:e01383902015. View Article : Google Scholar : PubMed/NCBI
44	Nikoletopoulou V, Markaki M, Palikaras K and Tavernarakis N: Crosstalk between apoptosis, necrosis and autophagy. Biochim Biophys Acta. 1833:3448–3459. 2013. View Article : Google Scholar : PubMed/NCBI
45	Okoshi R, Ozaki T, Yamamoto H, Ando K, Koida N, Ono S, Koda T, Kamijo T, Nakagawara A and Kizaki H: Activation of AMP-activated protein kinase induces p53-dependent apoptotic cell death in response to energetic stress. J Biol Chem. 283:3979–3987. 2008. View Article : Google Scholar : PubMed/NCBI
46	Kefas BA, Cai Y, Ling Z, Heimberg H, Hue L, Pipeleers D and Van de Casteele M: AMP-activated protein kinase can induce apoptosis of insulin-producing MIN6 cells through stimulation of c-Jun-N-terminal kinase. J Mol Endocrinol. 30:151–161. 2003. View Article : Google Scholar : PubMed/NCBI
47	Nieminen AI, Eskelinen VM, Haikala HM, Tervonen TA, Yan Y, Partanen JI and Klefström J: Myc-induced AMPK-phospho p53 pathway activates Bak to sensitize mitochondrial apoptosis. Proc Natl AcadSci USA. 110:pp. E1839–E1848. 2013; View Article : Google Scholar
48	Kim J, Kundu M, Viollet B and Guan KL: AMPK and mTOR regulate autophagy through direct phosphorylation of Ulk1. Nat Cell Biol. 13:132–141. 2011. View Article : Google Scholar : PubMed/NCBI
49	Jeon SM: Regulation and function of AMPK in physiology and diseases. Exp Mol Med. 48:e2452016. View Article : Google Scholar : PubMed/NCBI
50	Löffler AS, Alers S, Dieterle AM, Keppeler H, Franz-Wachtel M, Kundu M, Campbell DG, Wesselborg S, Alessi DR and Stork B: Ulk1-mediated phosphorylation of AMPK constitutes a negative regulatory feedback loop. Autophagy. 7:696–706. 2011. View Article : Google Scholar : PubMed/NCBI
51	Sanli T, Steinberg GR, Singh G and Tsakiridis T: AMP-activated protein kinase (AMPK) beyond metabolism: A novel genomic stress sensor participating in the DNA damage response pathway. Cancer Biol Ther. 15:156–169. 2014. View Article : Google Scholar : PubMed/NCBI
52	Kondo Y, Kanzawa T, Sawaya R and Kondo S: The role of autophagy in cancer development and response to therapy. Nat Rev Cancer. 5:726–734. 2005. View Article : Google Scholar : PubMed/NCBI
53	Xie BS, Zhao HC, Yao SK, Zhuo DX, Jin B, Lv DC, Wu CL, Ma DL, Gao C, Shu XM and Ai ZL: Autophagy inhibition enhances etoposide-induced cell death in human hepatoma G2 cells. Int J Mol Med. 27:599–606. 2011.PubMed/NCBI
54	Wu T, Wang MC, Jing L, Liu ZY, Guo H, Liu Y, Bai YY, Cheng YZ, Nan KJ and Liang X: Autophagy facilitates lung adenocarcinoma resistance to cisplatin treatment by activation of AMPK/mTOR signaling pathway. Drug Des Devel Ther. 9:6421–6431. 2015. View Article : Google Scholar : PubMed/NCBI