Combined antitumor effects of 1,25‑dihydroxy vitamin D3 and Notch inhibitor in liver cancer

Cai,Lijing; Luo,Lifang; Tang,Zhimou; Meng,Xiaohui

doi:10.3892/or.2018.6549

Combined antitumor effects of 1,25‑dihydroxy vitamin D3 and Notch inhibitor in liver cancer

Authors:
- Lijing Cai
- Lifang Luo
- Zhimou Tang
- Xiaohui Meng
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Affiliations: Department of Pharmacy, Jiangxi Provincial People's Hospital, Nanchang, Jiangxi 330006, P.R. China, Department of Pharmacy, Jiangxi Provincial People's Hospital, Nanchang, Jiangxi 330006, P.R. China, Department of Oncology, Jiangxi Provincial People's Hospital, Nanchang, Jiangxi 330006, P.R. China
Published online on: July 5, 2018 https://doi.org/10.3892/or.2018.6549
Pages: 1515-1524

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Abstract

The combined antitumor effects of 1,25‑dihydroxy vitamin D3 [1,25(OH)2D3] and the Notch inhibitor N‑[N‑(3,5‑difluorophenacetyl)‑L‑alanyl]‑S‑phenylglycine t‑butyl ester (DAPT, a synthetic γ secretase inhibitor) in liver cancer cells remain to be fully elucidated. In the present study, HepG2 cells were divided into six groups and different treatments were applied: Control, 10‑10 M 1,25(OH)2D3, 10‑8 M 1,25(OH)2D3, 10‑6 M 1,25(OH)2D3, 1 µM DAPT, 5 µM DAPT, 10 µM DAPT, and 10‑6 M 1,25(OH)2D3 + 10 µM DAPT. The proliferation, cell cycle, apoptosis, migration and invasion of the cells were then examined. The expression levels of Notch and its ligand Jagged were detected by reverse transcription‑quantitative polymerase chain reaction and western blot analyses. The results revealed that 1,25(OH)2D3 inhibited cell proliferation, migration and invasion; arrested cell cycle at the G1 phase, and promoted apoptosis in a concentration‑dependent manner between 10‑10 and 10‑6 M. DAPT inhibited cell proliferation, migration and invasion, arrested cell cycle at the G1 phase, and promoted apoptosis in a concentration‑dependent manner between 1 and 10 µM. Additionally, 1,25(OH)2D3 and/or DAPT reduced the expression of Notch1, Notch2, Jagged1 and Jagged2. The co‑application of 10 µM DAPT further increased the anticancer effect of 10‑6 M 1,25(OH)2D3. Collectively, these results indicated that the treatment of HepG2 cells with 1,25(OH)2D3 inactivated Notch signaling, prevented proliferation, migration and invasion, and promoted apoptosis. The combined application of 1,25(OH)2D3 with DAPT may be a useful treatment for preventing the onset or progression of liver cancer.

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1	Cevik O, Li D, Baljinnyam E, Manvar D, Pimenta EM, Waris G, Barnes BJ and Kaushik-Basu N: Interferon regulatory factor 5 (IRF5) suppresses hepatitis C virus (HCV) replication and HCV-associated hepatocellular carcinoma. J Biol Chem. 292:21676–21689. 2017. View Article : Google Scholar : PubMed/NCBI
2	Thein HH, Qiao Y, Zaheen A, Jembere N, Sapisochin G, Chan KK, Yoshida EM and Earle CC: Cost-effectiveness analysis of treatment with non-curative or palliative intent for hepatocellular carcinoma in the real-world setting. PLoS One. 12:e01851982017. View Article : Google Scholar : PubMed/NCBI
3	Ballotari P, Vicentini M, Manicardi V, Gallo M, Chiatamone Ranieri S, Greci M and Giorgi Rossi P: Diabetes and risk of cancer incidence: Results from a population-based cohort study in northern Italy. BMC Cancer. 17:7032017. View Article : Google Scholar : PubMed/NCBI
4	Kudo M, Kitano M, Sakurai T and Nishida N: General rules for the clinical and pathological study of primary liver cancer, nationwide follow-up survey and clinical practice guidelines: The outstanding achievements of the liver cancer study group of Japan. Dig Dis. 33:765–770. 2015. View Article : Google Scholar : PubMed/NCBI
5	Jia X, Gu Y, Groome LJ, Al-Kofahi M, Alexander JS, Li W and Wang Y: 1,25(OH)2D3 induces placental vascular smooth muscle cell relaxation by phosphorylation of myosin phosphatase target subunit 1^Ser507Potential beneficial effects of Vitamin D on placental vasculature in humans. Biol Reprod. 94:1162016. View Article : Google Scholar : PubMed/NCBI
6	Zhang CJ, Zhao D, Yin X, Zhang H, Ma L, Chen JP, Liu C and Yang XP: Effects of 1,25(OH)2D3 on proliferation and apoptosis of human glomerular mesangial cells. Am J Transl Res. 8:2659–2666. 2016.PubMed/NCBI
7	He XJ, Ding Y, Xiang W and Dang XQ: Roles of 1,25(OH)2D3 and Vitamin D receptor in the pathogenesis of rheumatoid arthritis and systemic lupus erythematosus by regulating the activation of CD4⁺ T cells and the PKCδ/ERK signaling pathway. Cell Physiol Biochem. 40:743–756. 2016. View Article : Google Scholar : PubMed/NCBI
8	Fazio C and Ricciardiello L: Inflammation and Notch signaling: A crosstalk with opposite effects on tumorigenesis. Cell Death Dis. 7:e25152016. View Article : Google Scholar : PubMed/NCBI
9	Luo J, Wang P, Wang R, Wang J, Liu M, Xiong S, Li Y and Cheng B: The Notch pathway promotes the cancer stem cell characteristics of CD90⁺ cells in hepatocellular carcinoma. Oncotarget. 7:9525–9537. 2016.PubMed/NCBI
10	Kim SJ, Lee HW, Baek JH, Cho YH, Kang HG, Jeong JS, Song J, Park HS and Chun KH: Activation of nuclear PTEN by inhibition of Notch signaling induces G2/M cell cycle arrest in gastric cancer. Oncogene. 35:251–260. 2016. View Article : Google Scholar : PubMed/NCBI
11	Tsai Y: Social security income and the utilization of home care: Evidence from the social security notch. J Health Econ. 43:45–55. 2015. View Article : Google Scholar : PubMed/NCBI
12	Li Z, Wang J, Zhao C, Ren K, Xia Z, Yu H and Jiang K: Acute blockage of Notch signaling by DAPT induces neuroprotection and neurogenesis in the neonatal rat brain after stroke. Transl Stroke Res. 7:132–140. 2016. View Article : Google Scholar : PubMed/NCBI
13	Wang J, Ye Z, Zheng S, Chen L, Wan Y, Deng Y and Yang R: Lingo-1 shRNA and Notch signaling inhibitor DAPT promote differentiation of neural stem/progenitor cells into neurons. Brain Res. 1634:34–44. 2016. View Article : Google Scholar : PubMed/NCBI
14	Jia M, Jiang L, Wang YD, Huang JZ, Yu M and Xue HZ: lincRNA-p21 inhibits invasion and metastasis of hepatocellular carcinoma through Notch signaling-induced epithelial-mesenchymal transition. Hepatol Res. 46:1137–1144. 2016. View Article : Google Scholar : PubMed/NCBI
15	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2^−ΔΔCT method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
16	Zeng N, Salker MS, Zhang S, Singh Y, Shi B, Stournaras C and Lang F: 1α,25(OH)2D3 Induces actin depolymerization in endometrial carcinoma cells by targeting RAC1 and PAK1. Cell Physiol Biochem. 40:1455–1464. 2016. View Article : Google Scholar : PubMed/NCBI
17	Osafi J, Hejazi A, Stutz DD, Keiserman MA, Bergman CJ and Kingsley K: Differential effects of 1,25-dihydroxyvitamin D3 on oral squamous cell carcinomas in vitro. J Diet Suppl. 11:145–154. 2014. View Article : Google Scholar : PubMed/NCBI
18	Lu HQ and Zheng J: Synergistic inhibitory effect of all-trans retinoic acid and 1,25-dihydroxy vitamin D3 on growth of human hepatoma cell line HepG2. Ai Zheng. 25:1470–1476. 2006.(In Chinese). PubMed/NCBI
19	Huang J, Yang G, Huang Y and Zhang S: Inhibitory effects of 1,25(OH)2D3 on the proliferation of hepatocellular carcinoma cells through the downregulation of HDAC2. Oncol Rep. 38:1845–1850. 2017. View Article : Google Scholar : PubMed/NCBI
20	Taniguchi K, Roberts LR, Aderca IN, Dong X, Qian C, Murphy LM, Nagorney DM, Burgart LJ, Roche PC, Smith DI, et al: Mutational spectrum of beta-catenin, AXIN1, and AXIN2 in hepatocellular carcinomas and hepatoblastomas. Oncogene. 21:48632002. View Article : Google Scholar : PubMed/NCBI
21	López-Terrada D, Cheung SW, Finegold MJ and Knowles BB: Hep G2 is a hepatoblastoma-derived cell line. Hum Pathol. 40:1512–1515. 2009. View Article : Google Scholar
22	Takiishi T, Ding L, Baeke F, Spagnuolo I, Sebastiani G, Laureys J, Verstuyf A, Carmeliet G, Dotta F, Van Belle TL, et al: Dietary supplementation with high doses of regular vitamin D3 safely reduces diabetes incidence in NOD mice when given early and long term. Diabetes. 63:2026–2036. 2014. View Article : Google Scholar : PubMed/NCBI
23	Tangpricha V, Spina C, Yao M, Chen TC, Wolfe MM and Holick MF: Vitamin D deficiency enhances the growth of MC-26 colon cancer xenografts in Balb/c mice. J Nutr. 135:2350–2354. 2005. View Article : Google Scholar : PubMed/NCBI
24	Huang J, Yang G, Huang Y and Zhang S: 1,25(OH)2D3 induced apoptosis of human hepatocellular carcinoma cells in vitro and inhibited their growth in a nude mouse xenograft model by regulating histone deacetylase 2. Biochimie. 146:28–34. 2018. View Article : Google Scholar : PubMed/NCBI
25	Li M, Li L, Zhang L, Hu W, Shen J, Xiao Z, Wu X, Chan FL and Cho CH: 1,25-Dihydroxyvitamin D3 suppresses gastric cancer cell growth through VDR- and mutant p53-mediated induction of p21. Life Sci. 179:88–97. 2017. View Article : Google Scholar : PubMed/NCBI
26	Lin YC, Lee BH, Alagie J and Su CH: Combination treatment of ergosterol followed by amphotericin B induces necrotic cell death in human hepatocellular carcinoma cells. Oncotarget. 8:72727–72738. 2017.PubMed/NCBI
27	Liu X, Sheng HB, Ma R, Yang JM, Luo WW, Yang XY, Ren DD and Chi FL: Notch signaling is active in normal mouse middle ear epithelial cells. Exp Ther Med. 11:1661–1667. 2016. View Article : Google Scholar : PubMed/NCBI
28	Wang Q, Chen X and Hay N: Akt as a target for cancer therapy: More is not always better (lessons from studies in mice). Br J Cancer. 117:159–163. 2017. View Article : Google Scholar : PubMed/NCBI
29	de Melo AC, Paulino E and Garces ÁH: A review of mTOR pathway inhibitors in gynecologic cancer. Oxid Med Cell Longev. 2017:48097512017. View Article : Google Scholar : PubMed/NCBI
30	Murta D, Batista M, Silva E, Trindade A, Henrique D, Duarte A and Lopes-da-Costa L: Notch signaling in the epididymal epithelium regulates sperm motility and is transferred at a distance within epididymosomes. Andrology. 4:314–327. 2016. View Article : Google Scholar : PubMed/NCBI
31	Abad M, Hashimoto H, Zhou H, Morales MG, Chen B, Bassel-Duby R and Olson EN: Notch inhibition enhances cardiac reprogramming by increasing MEF2C transcriptional activity. Stem Cell Reports. 8:548–560. 2017. View Article : Google Scholar : PubMed/NCBI
32	Zhao D, Zhang CJ, Yang R, Chen JP, Ma L, Liu G and Yang XP: Effect of 1,25(OH2D3 on the proliferation of human mesangial cells and their expression of Ki67. Genet Mol Res. 16:2017.doi: 10.4238/gmr16029191. View Article : Google Scholar
33	Song JH, Park E, Kim MS, Cho KM, Park SH, Lee A, Song J, Kim HJ, Koh JT and Kim TS: l-Asparaginase-mediated downregulation of c-Myc promotes 1,25(OH)2 D3-induced myeloid differentiation in acute myeloid leukemia cells. Int J Cancer. 140:2364–2374. 2017. View Article : Google Scholar : PubMed/NCBI
34	Abu El Maaty MA, Alborzinia H, Khan SJ, Buttner M and Wölfl S: 1,25(OH)2D3 disrupts glucose metabolism in prostate cancer cells leading to a truncation of the TCA cycle and inhibition of TXNIP expression. Biochim Biophys Acta. 1864:1618–1630. 2017. View Article : Google Scholar : PubMed/NCBI
35	Fesik SW and Shi Y: Structural biology. Controlling the caspases. Science. 294:1477–1478. 2001. View Article : Google Scholar : PubMed/NCBI
36	Stacey DW: Cyclin D1 serves as a cell cycle regulatory switch in actively proliferating cells. Curr Opin Cell Biol. 15:158–163. 2003. View Article : Google Scholar : PubMed/NCBI
37	Geisler F and Strazzabosco M: Emerging roles of Notch signaling in liver disease. Hepatology. 61:382–392. 2015. View Article : Google Scholar : PubMed/NCBI
38	Xu J, Chi F, Guo T, Punj V, Lee WN, French SW and Tsukamoto H: NOTCH reprograms mitochondrial metabolism for proinflammatory macrophage activation. J Clin Invest. 125:1579–1590. 2015. View Article : Google Scholar : PubMed/NCBI
39	D'Angelo RC, Ouzounova M, Davis A, Choi D, Tchuenkam SM, Kim G, Luther T, Quraishi AA, Senbabaoglu Y, Conley SJ, et al: Notch reporter activity in breast cancer cell lines identifies a subset of cells with stem cell activity. Mol Cancer Ther. 14:779–787. 2015. View Article : Google Scholar : PubMed/NCBI
40	Song BQ, Chi Y, Li X, Du WJ, Han ZB, Tian JJ, Li JJ, Chen F, Wu HH, Han LX, et al: Inhibition of Notch signaling promotes the adipogenic differentiation of mesenchymal stem cells through autophagy activation and PTEN-PI3K/AKT/mTOR pathway. Cell Physiol Biochem. 36:1991–2002. 2015. View Article : Google Scholar : PubMed/NCBI