Suppression of 14-3-3ζ in cholangiocarcinoma cells inhibits proliferation through attenuated Akt activity, enhancing chemosensitivity to gemcitabine

Kittirat,Yingpinyapat; Techasen,Anchalee; Thongchot,Suyanee; Loilome,Watcharin; Thanan,Raynoo; Yongvanit,Puangrat; Sungkhamanon,Sakkarn; Titapun,Attapol; Khuntikeo,Narong; Namwat,Nisana

doi:10.3892/ol.2017.7326

Suppression of 14-3-3ζ in cholangiocarcinoma cells inhibits proliferation through attenuated Akt activity, enhancing chemosensitivity to gemcitabine

Authors:
- Yingpinyapat Kittirat
- Anchalee Techasen
- Suyanee Thongchot
- Watcharin Loilome
- Raynoo Thanan
- Puangrat Yongvanit
- Sakkarn Sungkhamanon
- Attapol Titapun
- Narong Khuntikeo
- Nisana Namwat
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Affiliations: Department of Biochemistry, Faculty of Medicine, Khon Kaen University, Khon Kaen, Muang 40002, Thailand, Cholangiocarcinoma Research Institute, Khon Kaen University, Khon Kaen, Muang 40002, Thailand
Published online on: November 1, 2017 https://doi.org/10.3892/ol.2017.7326
Pages: 347-353
Copyright: © Kittirat et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The protein 14-3-3ζ contributes important regulatory functions in several cellular processes via binding to phosphorylated serine/threonine residues, which promotes cell cycle progression, cell proliferation and anti‑apoptosis in multiple types of cancer. The aim of the present study was to investigate the functions of 14‑3‑3ζ in cholangiocarcinoma (CCA) progression and elucidate the molecular mechanism of 14‑3‑3ζ expression‑mediated protein kinase B (Akt) phosphorylation and chemosensitivity in CCA cells. In the present study, 14‑3‑3ζ expression was investigated in clinical specimens using immunohistochemistry and compared with the clinicopathological features of patients with CCA. The association between 14‑3‑3ζ and phosphorylated Akt (pAkt) was determined among the tissues of the same patients using bivariate correlation analysis. The effects of 14‑3‑3ζ suppression on CCA cell function and gemcitabine sensitivity were investigated using small interfering RNA (siRNA). It was identified that 14‑3‑3ζ expression was positively correlated with pAkt (P=0.013) and that increased expression of 14‑3‑3ζ and pAkt were significantly associated with poor overall survival rate and metastasis (P=0.025 and 0.006, respectively). Downregulation of 14‑3‑3ζ using siRNA in CCA cell lines decreased cell proliferation, resulting in the inhibition of pAkt activity and increasing the protein level of the cell cycle inhibitor p27. The suppression of 14‑3‑3ζ enhanced the inhibitory effect of gemcitabine on CCA cell proliferation by inducing apoptotic cell death. Taken together, the results of the present study indicated that 14‑3‑3ζ is a potential target for CCA and may serve as a novel therapeutic approach to enhance chemosensitivity in the treatment of CCA.

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1	Khan SA, Emadossadaty S, Ladep NG, Thomas HC, Elliott P, Taylor-Robinson SD and Toledano MB: Rising trends in cholangiocarcinoma: Is the ICD classification system misleading us? J Hepatol. 56:848–854. 2012. View Article : Google Scholar : PubMed/NCBI
2	Bertuccio P, Bosetti C, Levi F, Decarli A, Negri E and La Vecchia C: A comparison of trends in mortality from primary liver cancer and intrahepatic cholangiocarcinoma in Europe. Ann Oncol. 24:1667–1674. 2013. View Article : Google Scholar : PubMed/NCBI
3	Ghouri YA, Mian I and Blechacz B: Cancer review: Cholangiocarcinoma. J Carcinog. 14:12015. View Article : Google Scholar : PubMed/NCBI
4	Sripa B and Pairojkul C: Cholangiocarcinoma: Lessons from Thailand. Curr Opin Gastroenterol. 24:349–356. 2008. View Article : Google Scholar : PubMed/NCBI
5	Sriamporn S, Pisani P, Pipitgool V, Suwanrungruang K, Kamsa-ard S and Parkin DM: Prevalence of Opisthorchis viverrini infection and incidence of cholangiocarcinoma in Khon Kaen, Northeast Thailand. Trop Med Int Health. 9:588–594. 2004. View Article : Google Scholar : PubMed/NCBI
6	Bundhamcharoen K, Odton P, Phulkerd S and Tangcharoensathien V: Burden of disease in Thailand: Changes in health gap between 1999 and 2004. BMC Public Health. 11:532011. View Article : Google Scholar : PubMed/NCBI
7	Valle JW, Wasan H, Johnson P, Jones E, Dixon L, Swindell R, Baka S, Maraveyas A, Corrie P, Falk S, et al: Gemcitabine alone or in combination with cisplatin in patients with advanced or metastatic cholangiocarcinomas or other biliary tract tumours: A multicentre randomised phase II study-The UK ABC-01 Study. Br J Cancer. 101:621–627. 2009. View Article : Google Scholar : PubMed/NCBI
8	Freeman AK and Morrison DK: 14-3-3 Proteins: Diverse functions in cell proliferation and cancer progression. Semin Cell Dev Biol. 22:681–687. 2011. View Article : Google Scholar : PubMed/NCBI
9	Neal CL, Yao J, Yang W, Zhou X, Nguyen NT, Lu J, Danes CG, Guo H, Lan KH, Ensor J, et al: 14-3-3zeta overexpression defines high risk for breast cancer recurrence and promotes cancer cell survival. Cancer Res. 69:3425–3432. 2009. View Article : Google Scholar : PubMed/NCBI
10	Matta A, Bahadur S, Duggal R, Gupta SD and Ralhan R: Over-expression of 14-3-3zeta is an early event in oral cancer. BMC Cancer. 7:1692007. View Article : Google Scholar : PubMed/NCBI
11	Nishimura Y, Komatsu S, Ichikawa D, Nagata H, Hirajima S, Takeshita H, Kawaguchi T, Arita T, Konishi H, Kashimoto K, et al: Overexpression of YWHAZ relates to tumor cell proliferation and malignant outcome of gastric carcinoma. Br J Cancer. 108:1324–1331. 2013. View Article : Google Scholar : PubMed/NCBI
12	Bajpai U, Sharma R, Kausar T, Dattagupta S, Chattopadhayay TK and Ralhan R: Clinical significance of 14-3-3 zeta in human esophageal cancer. Int J Biol Markers. 23:231–237. 2008. View Article : Google Scholar : PubMed/NCBI
13	Zhang C, Liu LX, Dong ZR, Shi GM, Cai JB, Zhang PF, Ke AW, Yu JX, Zhou J and Fan J: Up-regulation of 14-3-3zeta expression in intrahepatic cholangiocarcinoma and its clinical implications. Tumour Biol. 36:1781–1789. 2015. View Article : Google Scholar : PubMed/NCBI
14	Frasor J, Chang EC, Komm B, Lin CY, Vega VB, Liu ET, Miller LD, Smeds J, Bergh J and Katzenellenbogen BS: Gene expression preferentially regulated by tamoxifen in breast cancer cells and correlations with clinical outcome. Cancer Res. 66:7334–7340. 2006. View Article : Google Scholar : PubMed/NCBI
15	Fan T, Li R, Todd NW, Qiu Q, Fang HB, Wang H, Shen J, Zhao RY, Caraway NP, Katz RL, et al: Up-regulation of 14-3-3zeta in lung cancer and its implication as prognostic and therapeutic target. Cancer Res. 67:7901–7906. 2007. View Article : Google Scholar : PubMed/NCBI
16	Neal CL, Xu J, Li P, Mori S, Yang J, Neal NN, Zhou X, Wyszomierski SL and Yu D: Overexpression of 14-3-3zeta in cancer cells activates PI3K via binding the p85 regulatory subunit. Oncogene. 31:897–906. 2012. View Article : Google Scholar : PubMed/NCBI
17	Yothaisong S, Dokduang H, Techasen A, Namwat N, Yongvanit P, Bhudhisawasdi V, Puapairoj A, Riggins GJ and Loilome W: Increased activation of PI3K/AKT signaling pathway is associated with cholangiocarcinoma metastasis and PI3K/mTOR inhibition presents a possible therapeutic strategy. Tumour Biol. 34:3637–3648. 2013. View Article : Google Scholar : PubMed/NCBI
18	Matta A, Siu KW and Ralhan R: 14-3-3zeta as novel molecular target for cancer therapy. Expert Opin Ther Targets. 16:515–523. 2012. View Article : Google Scholar : PubMed/NCBI
19	Matta A, DeSouza LV, Shukla NK, Gupta SD, Ralhan R and Siu KW: Prognostic significance of head-and-neck cancer biomarkers previously discovered and identified using iTRAQ-labeling and multidimensional liquid chromatography-tandem mass spectrometry. J Proteome Res. 7:2078–2087. 2008. View Article : Google Scholar : PubMed/NCBI
20	Khan SA, Thomas HC, Davidson BR and Taylor-Robinson SD: Cholangiocarcinoma. Lancet. 366:1303–1314. 2005. View Article : Google Scholar : PubMed/NCBI