TPX2 silencing exerts anti‑tumor effects on hepatocellular carcinoma by regulating the PI3K/AKT signaling pathway

Huang,Dan‑Hong; Jian,Jie; Li,Shuang; Zhang,Yan; Liu,Li‑Zhen

doi:10.3892/ijmm.2019.4371

TPX2 silencing exerts anti‑tumor effects on hepatocellular carcinoma by regulating the PI3K/AKT signaling pathway

Authors:
- Dan‑Hong Huang
- Jie Jian
- Shuang Li
- Yan Zhang
- Li‑Zhen Liu
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Affiliations: Department of Clinical Laboratory, Jiading District Central Hospital of Shanghai University of Medicine and Health Sciences, Shanghai 201800, P.R. China, Department of Gastroenterology, First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China, Department of Gastroenterology, Third Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330008, P.R. China, Department of Oncology, Jiading District Central Hospital of Shanghai University of Medicine and Health Sciences, Shanghai 201800, P.R. China
Published online on: October 14, 2019 https://doi.org/10.3892/ijmm.2019.4371
Pages: 2113-2122
Copyright: © Huang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Hepatocellular carcinoma (HCC) is one of the primary causes of cancer‑associated deaths worldwide. Current treatment methods include surgical resection, chemotherapy and radiotherapy; however the curative rate remains low, thus novel treatments are required. The aim of the present study was to investigate the role of targeting protein for Xenopus kinesin‑like protein 2 (TPX2) in the growth of HCC and its underlying molecular mechanism. Immunohistochemistry staining, reverse transcription‑quantitative (RT‑q)PCR and western blotting were used to detect the expression of TPX2 mRNA and protein in liver cancer tissue samples, adjacent normal liver tissue samples, and the HCC cell lines Huh7, Hep3B, PLC/PRF/5 and MHCC97‑H. The recombinant plasmid pMagic4.1‑shRNA‑TPX2 was constructed and transfected into Huh7 and Hep3B HCC cells to silence TPX2 expression. The proliferation, apoptosis, migration and invasion of Huh7 cells and Hep3B cells were evaluated before and after TPX2 silencing. The mRNA and protein expression levels of multiple signaling pathway‑associated genes were detected by RT‑qPCR and western blotting. The expression levels of TPX2 mRNA and protein were significantly higher in HCC tissue samples compared with adjacent normal liver tissue sample. TPX2 mRNA and protein expression levels were detected in the different HCC cell lines. The recombinant plasmid pMagic4.1‑shRNA‑TPX2 was successfully transfected into Huh7 and Hep3B cells, resulting in TPX2 silencing. TPX2 knockdown significantly reduced cell proliferation, cell migration and cell invasion of Huh7 and Hep3B cells, whilst also increasing the rate of apoptosis in these cells. Following TPX2 silencing, the expression levels of PI3K, phospho‑AKT, Bcl‑2, c‑Myc and Cyclin D1 were significantly decreased, whereas the expression levels of P21 and P27 were significantly increased. In conclusion, TPX2 may suppress the growth of HCC by regulating the PI3K/AKT signaling pathway and thus, TPX2 may be a potential target for the treatment of liver cancer.

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1	Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA and Jemal A: Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 68:394–424. 2018. View Article : Google Scholar : PubMed/NCBI
2	Dandri M and Petersen J: Mechanism of hepatitis B virus persistence in hepatocytes and its carcinogenic potential. Clin Infect Dis. 62(Suppl 4): S281–S288. 2016. View Article : Google Scholar : PubMed/NCBI
3	Wang M, Wang Y, Feng X, Wang R, Wang Y, Zeng H, Qi J, Zhao H, Li N, Cai J and Qu C: Contribution of hepatitis B virus and hepatitis C virus to liver cancer in China north areas: Experience of the Chinese National Cancer Center. Int J Infect Dis. 65:15–21. 2017. View Article : Google Scholar : PubMed/NCBI
4	de Martel C, Maucort-Boulch D, Plummer M and Franceschi S: World-wide relative contribution of hepatitis B and C viruses in hepatocellular carcinoma. Hepatology. 62:1190–1200. 2015. View Article : Google Scholar : PubMed/NCBI
5	Younossi ZM, Otgonsuren M, Henry L, Venkatesan C, Mishra A, Erario M and Hunt S: Association of nonalcoholic fatty liver disease (NAFLD) with hepatocellular carcinoma (HCC) in the United States from 2004 to 2009. Hepatology. 62:1723–1730. 2015. View Article : Google Scholar : PubMed/NCBI
6	Liu Z, Jiang Y, Yuan H, Fang Q, Cai N, Suo C, Jin L, Zhang T and Chen X: The trends in incidence of primary liver cancer caused by specific etiologies: Results from the Global Burden of Disease Study 2016 and implications for liver cancer prevention. J Hepatol. 70:674–683. 2019. View Article : Google Scholar
7	Gruss OJ and Vernos I: The mechanism of spindle assembly: Functions of Ran and its target TPX2. J Cell Biol. 166:949–955. 2004. View Article : Google Scholar : PubMed/NCBI
8	Rennie YK, McIntyre PJ, Akindele T, Bayliss R and Jamieson AG: A TPX2 proteomimetic has enhanced affinity for Aurora-A due to hydrocarbon stapling of a Helix. ACS Chem Biol. 11:3383–3390. 2016. View Article : Google Scholar : PubMed/NCBI
9	Pascreau G, Eckerdt F, Lewellyn AL, Prigent C and Maller JL: Phosphorylation of p53 is regulated by TPX2-Aurora A in xenopus oocytes. J Biol Chem. 284:5497–5505. 2009. View Article : Google Scholar : PubMed/NCBI
10	Liu HC, Zhang Y, Wang XL, Qin WS, Liu YH, Zhang L and Zhu CL: Upregulation of the TPX2 gene is associated with enhanced tumor malignance of esophageal squamous cell carcinoma. Biomed Pharmacother. 67:751–755. 2013. View Article : Google Scholar : PubMed/NCBI
11	Takahashi Y, Sheridan P, Niida A, Sawada G, Uchi R, Mizuno H, Kurashige J, Sugimachi K, Sasaki S, Shimada Y, et al: The AURKA/TPX2 axis drives colon tumorigenesis cooperatively with MYC. Ann Oncol. 26:935–942. 2015. View Article : Google Scholar : PubMed/NCBI
12	Wei P, Zhang N, Xu Y, Li X, Shi D, Wang Y, Li D and Cai S: TPX2 is a novel prognostic marker for the growth and metastasis of colon cancer. J Transl Med. 11:3132013. View Article : Google Scholar : PubMed/NCBI
13	Chen M, Zhang H, Zhang G, Zhong A, Ma Q, Kai J, Tong Y, Xie S, Wang Y, Zheng H, et al: Targeting TPX2 suppresses proliferation and promotes apoptosis via repression of the PI3k/AKT/P21 signaling pathway and activation of p53 pathway in breast cancer. Biochem Biophys Res Commun. 507:74–82. 2018. View Article : Google Scholar : PubMed/NCBI
14	Jiang P, Shen K, Wang X, Song H, Yue Y and Liu T: TPX2 regulates tumor growth in human cervical carcinoma cells. Mol Med Rep. 9:2347–2351. 2014. View Article : Google Scholar : PubMed/NCBI
15	Chang H, Wang J, Tian Y, Xu J, Gou X and Cheng J: The TPX2 gene is a promising diagnostic and therapeutic target for cervical cancer. Oncol Rep. 27:1353–1359. 2012.PubMed/NCBI
16	Tian Y, Liu LL, Guo DM, Wang Y, Zha WH, Li Y and Wu FJ: TPX2 gene silencing inhibits cell proliferation and promotes apoptosis through negative regulation of AKT signaling pathway in ovarian cancer. J Cell Biochem. 119:7540–7555. 2018. View Article : Google Scholar : PubMed/NCBI
17	Yan L, Li Q, Yang J and Qiao B: TPX2-p53-GLIPR1 regulatory circuitry in cell proliferation, invasion, and tumor growth of bladder cancer. J Cell Biochem. 119:1791–1803. 2018. View Article : Google Scholar
18	Yang X, Liu G, Xiao H, Yu F, Xiang X, Lu Y, Li W, Liu X, Li S and Shi Y: TPX2 overexpression in medullary thyroid carcinoma mediates TT cell proliferation. Pathol Oncol Res. 20:641–648. 2014. View Article : Google Scholar : PubMed/NCBI
19	Jian J, Huang Y, Liu LZ, Li SX and Deng F: TPX2 gene-silencing inhibits the proliferation and invasion of human colon cancer SW480 cells. TUMOR. 36:628–634. 2016.
20	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T) method. Methods. 25:402–408. 2001. View Article : Google Scholar
21	Llovet JM, Bruix J, Fuster J, Castells A, Garcia-Valdecasas JC, Grande L, Franca A, Brú C, Navasa M, Ayuso MC, et al: Liver transplantation for small hepatocellular carcinoma: The tumor-node-metastasis classification does not have prognostic power. Hepatology. 27:1572–1577. 1998. View Article : Google Scholar : PubMed/NCBI
22	Xue S, Zhou Y, Zhang J, Xiang Z, Liu Y, Miao T, Liu G, Liu B, Liu X, Shen L, et al: Anemoside B4 exerts anti-cancer effect by inducing apoptosis and autophagy through inhibiton of PI3K/Akt/mTOR pathway in hepatocellular carcinoma. Am J Transl Res. 11:2580–2589. 2019.PubMed/NCBI
23	Feng PC, Ke XF, Kuang HL, Pan LL, Ye Q and Wu JB: BMP2 secretion from hepatocellular carcinoma cell HepG2 enhances angiogenesis and tumor growth in endothelial cells via activation of the MAPK/p38 signaling pathway. Stem Cell Res Ther. 10:2372019. View Article : Google Scholar : PubMed/NCBI
24	Li SJ, Sui MH, Sun ZX and Zhang WW: LncRNA 00152 promotes the development of hepatocellular carcinoma by activating JAK2/STAT3 pathway. Eur Rev Med Pharmacol Sci. 23:1038–1046. 2019.PubMed/NCBI
25	Zuo J, Ma H, Cai H, Wu Y, Jiang W and Yu L: An inhibitory role of NEK6 in TGFβ/Smad signaling pathway. BMB Rep. 48:473–478. 2015. View Article : Google Scholar :
26	Yang Y, Yang X, Li L, Yang G, Ouyang X, Xiang J, Zhang T and Min X: LASS2 inhibits proliferation and induces apoptosis in HepG2 cells by affecting mitochondrial dynamics, the cell cycle and the nuclear factor-κB pathways. Oncol Rep. 41:3005–3014. 2019.PubMed/NCBI
27	Thelen A, Benckert C, Tautenhahn HM, Hau HM, Bartels M, Linnemann J, Bertolini J, Moche M, Wittekind C and Jonas S: Liver resection for hepatocellular carcinoma in patients without cirrhosis. Br J Surg. 100:130–137. 2013. View Article : Google Scholar
28	Rhu J, Kim JM, Choi GS, Kwon CHD and Joh JW: Continuing five or more locoregional therapies before living donor salvage liver transplantation for hepatocellular carcinoma is related to poor recurrence-free survival. Ann Surg Treat Res. 95:152–160. 2018. View Article : Google Scholar : PubMed/NCBI
29	Anderson TN and Zarrinpar A: Hepatocyte transplantation: Past efforts, current technology, and future expansion of therapeutic potential. J Surg Res. 226:48–55. 2018. View Article : Google Scholar : PubMed/NCBI
30	Famularo S, Di Sandro S, Giani A, Lauterio A, Sandini M, De Carlis R, Buscemi V, Uggeri F, Romano F, Gianotti L and De Carlis L: Recurrence patterns after anatomic or paren-chyma-sparing liver resection for hepatocarcinoma in a western population of cirrhotic patients. Ann Surg Oncol. 25:3974–3981. 2018. View Article : Google Scholar : PubMed/NCBI
31	Meischl T, Rasoul-Rockenschaub S, Györi G, Sieghart W, Reiberger T, Trauner M, Soliman T, Berlakovich G and Pinter M: C-reactive protein is an independent predictor for hepatocellular carcinoma recurrence after liver transplantation. PLoS One. 14:e02166772019. View Article : Google Scholar : PubMed/NCBI
32	Scaggiante B, Kazemi M, Pozzato G, Dapas B, Farra R, Grassi M, Zanconati F and Grassi G: Novel hepatocellular carcinoma molecules with prognostic and therapeutic potentials. World J Gastroenterol. 20:1268–1288. 2014. View Article : Google Scholar : PubMed/NCBI
33	Gao B, Li S, Tan Z, Ma L and Liu J: ACTG1 and TLR3 are biomarkers for alcohol-associated hepatocellular carcinoma. Oncol Lett. 17:1714–1722. 2019.PubMed/NCBI
34	Augello G, Emma MR, Cusimano A, Azzolina A, Mongiovì S, Puleio R, Cassata G, Gulino A, Belmonte B, Gramignoli R, et al: Targeting HSP90 with the small molecule inhibitor AUY922 (luminespib) as a treatment strategy against hepatocellular carcinoma. Int J Cancer. 144:2613–2624. 2019. View Article : Google Scholar
35	Pan W, Luo Q, Yan X, Yuan L, Yi H, Zhang L, Li B, Zhang Y, Sun J, Qiu MZ and Yang DJ: A novel SMAC mimetic APG-1387 exhibits dual antitumor effect on HBV-positive hepatocellular carcinoma with high expression of cIAP2 by inducing apoptosis and enhancing innate anti-tumor immunity. Biochem Pharmacol. 154:127–135. 2018. View Article : Google Scholar : PubMed/NCBI
36	Neumayer G, Belzil C, Gruss OJ and Nguyen MD: TPX2: Of spindle assembly, DNA damage response, and cancer. Cell Mol Life Sci. 71:3027–3047. 2014. View Article : Google Scholar : PubMed/NCBI
37	Wittmann T, Wilm M, Karsenti E and Vernos I: TPX2, A novel xenopus MAP involved in spindle pole organization. J Cell Biol. 149:1405–1418. 2000. View Article : Google Scholar : PubMed/NCBI
38	Moss DK, Wilde A and Lane JD: Dynamic release of nuclear RanGTP triggers TPX2-dependent microtubule assembly during the apoptotic execution phase. J Cell Sci. 122:644–655. 2009. View Article : Google Scholar : PubMed/NCBI
39	Zhang H, Pan YZ, Cheung M, Cao M, Yu C, Chen L, Zhan L, He ZW and Sun CY: LAMB3 mediates apoptotic, proliferative, invasive, and metastatic behaviors in pancreatic cancer by regulating the PI3K/Akt signaling pathway. Cell Death Dis. 10:2302019. View Article : Google Scholar : PubMed/NCBI
40	Sun Y, Cao FL, Qu LL, Wang ZM and Liu XY: MEG3 promotes liver cancer by activating PI3K/AKT pathway through regulating AP1G1. Eur Rev Med Pharmacol Sci. 23:1459–1467. 2019.PubMed/NCBI
41	Cen D, Huang H, Yang L, Guo K and Zhang J: Long noncoding RNA STXBP5-AS1 inhibits cell proliferation, migration, and invasion through inhibiting the PI3K/AKT signaling pathway in gastric cancer cells. Onco Targets Ther. 12:1929–1936. 2019. View Article : Google Scholar : PubMed/NCBI
42	Yun WK, Hu YM, Zhao CB, Yu DY and Tang JB: HCP5 promotes colon cancer development by activating AP1G1 via PI3K/AKT pathway. Eur Rev Med Pharmacol Sci. 23:2786–2793. 2019.PubMed/NCBI
43	Li H, Zhang Q, Wu Q, Cui Y, Zhu H, Fang M, Zhou X, Sun Z and Yu J: Interleukin-22 secreted by cancer-associated fibroblasts regulates the proliferation and metastasis of lung cancer cells via the PI3K-Akt-mTOR signaling pathway. Am J Transl Res. 11:4077–4088. 2019.PubMed/NCBI
44	Chen T, Gu C, Xue C, Yang T, Zhong Y, Liu S, Nie Y and Yang H: LncRNA-uc002mbe.2 interacting with hnRNPA2B1 mediates AKT deactivation and p21 up-regulation induced by trichostatin in liver cancer cells. Front Pharmacol. 8:6692017. View Article : Google Scholar : PubMed/NCBI
45	Chen T, Huang H, Zhou Y, Geng L, Shen T, Yin S, Zhou L and Zheng S: HJURP promotes hepatocellular carcinoma proliferation by destabilizing p21 via the MAPK/ERK1/2 and AKT/GSK3β signaling pathways. J Exp Clin Cancer Res. 37:1932018. View Article : Google Scholar
46	Zhang Y, Liu Y, Duan J, Yan H, Zhang J, Zhang H, Fan Q, Luo F, Yan G, Qiao K and Liu J: Hippocalcin-like 1 suppresses hepatocellular carcinoma progression by promoting p21(Waf/Cip1) stabilization by activating the ERK1/2-MAPK pathway. Hepatology. 63:880–897. 2016. View Article : Google Scholar
47	Ohkoshi S, Yano M and Matsuda Y: Oncogenic role of p21 in hepatocarcinogenesis suggests a new treatment strategy. World J Gastroenterol. 21:12150–12156. 2015. View Article : Google Scholar : PubMed/NCBI
48	Ma J, Ren Y, Zhang L, Kong X, Wang T, Shi Y and Bu R: Knocking-down of CREPT prohibits the progression of oral squamous cell carcinoma and suppresses cyclin D1 and c-Myc expression. PLoS One. 12:e01743092017. View Article : Google Scholar : PubMed/NCBI
49	Chen Y, Fang L, Zhang J, Li G, Ma M, Li C, Lyu J and Meng QH: Blockage of Glyoxalase I inhibits colorectal tumorigenesis and tumor growth via upregulation of STAT1, p53, and Bax and downregulation of c-Myc and Bcl-2. Int J Mol Sci. 18:2017.