RNA interference‑mediated knockdown of translationally controlled tumor protein induces apoptosis, and inhibits growth and invasion in glioma cells

Jin,Hua; Zhang,Xuexin; Su,Jun; Teng,Yueqiu; Ren,Huan; Yang,Lizhuang

doi:10.3892/mmr.2015.4280

RNA interference‑mediated knockdown of translationally controlled tumor protein induces apoptosis, and inhibits growth and invasion in glioma cells

Authors:
- Hua Jin
- Xuexin Zhang
- Jun Su
- Yueqiu Teng
- Huan Ren
- Lizhuang Yang
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Affiliations: Department of Immunology, Harbin Medical University, Harbin, Heilongjiang 150081, P.R. China, Department of Neurosurgery, The Third Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150040, P.R. China, Stem Cell Research Institute, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001, P.R. China, Department of Neurosurgery, The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150081, P.R. China
Published online on: September 2, 2015 https://doi.org/10.3892/mmr.2015.4280
Pages: 6617-6625
Copyright: © Jin et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Translationally controlled tumor protein (TCTP) is a highly conserved, growth‑associated and small molecule protein, which is highly expressed in various types of tumor cell. TCTP can promote the growth and suppress apoptosis of tumor cels. However, few studies have reported the effects of TCTP in gliomas. In the present study, a glioma cell line was established, which was stably transfected with TCTP short hairpin ribonucleic acid (shRNA), to investigate the impact of downregulated expression of TCTP on the proliferation, apoptosis and invasion of glioma cells. Western blot and reverse transcription-quantitative polymerase chain reaction analyses demonstrated that TCTP shRNA effectively reduced the expression of TCTP in the U251 glioma cell line. MTT and colony formation assays revealed that downregulated expression of TCTP significantly inhibited glioma cell proliferation. Cell cycle analysis using flow cytometry revealed that the cells in the pRNA‑H1.1‑TCTP group were arrested in the G0/G1 phase of the cell cycle. Western blot analysis detected downregulated expression levels of cyclins, including Cyclin D1, Cyclin E and Cyclin B. Annexin V‑fluorescein isothiocyanate/propidium iodide and Hoechst staining demonstrated that the apoptotic rate of the cells in the pRNA‑H1.1‑TCTP group was significantly higher than that of the cells in the pRNA‑H1.1‑control group, with upregulated expression levels of B-cell-associated X protein and cleaved‑caspase‑3 and downregulated expression of B-cell lmyphoma-2 in the apoptotic process. Wound healing and Transwell assays revealed that downregulated expression of TCTP significantly inhibited the migration and invasiveness of the glioma cells; and the expression levels and activities of matrix metalloproteinase (MMP)‑2 and MMP‑9 were also significantly affected. In conclusion, the present study demonstrated that downregulated expression of TCTP significantly inhibited proliferation and invasion, and induced apoptosis in the glioma cells. These results suggested that TCTP may be important in glioma development and metastasis. Therefore, TCTP is expected to become an effective target for glioma gene therapy.

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1	Fine HA, Dear KB, Loeffler JS, Black PM and Canellos GP: Meta-analysis of radiation therapy with and without adjuvant chemotherapy for malignant gliomas in adults. Cancer. 71:2585–2597. 1993. View Article : Google Scholar : PubMed/NCBI
2	Wu X, Li Y, Wan X, Kayira TM, Cao R, Ju X, Zhu X and Zhao G: Down-regulation of neogenin accelerated glioma progression through promoter Methylation and its overexpression in SHG-44 Induced Apoptosis. PLoS One. 7:e380742012. View Article : Google Scholar : PubMed/NCBI
3	Van Meir EG, Hadjipanayis CG, Norden AD, Shu HK, Wen PY and Olson JJ: Exciting new advances in neuro-oncology: The avenue to a cure for malignant glioma. CA Cancer J Clin. 60:166–193. 2010. View Article : Google Scholar : PubMed/NCBI
4	Turbyville TJ, Gürsel DB, Tuskan RG, Walrath JC, Lipschultz CA, Lockett SJ, Wiemer DF, Beutler JA and Reilly KM: Schweinfurthin A selectively inhibits proliferation and Rho signaling in glioma and neurofibromatosis type 1 tumor cells in a NF1-GRD-dependent manner. Mol Cancer Ther. 9:1234–1243. 2010. View Article : Google Scholar : PubMed/NCBI
5	Ohgaki H and Kleihues P: Epidemiology and etiology of gliomas. Acta Neuropathol. 109:93–108. 2005. View Article : Google Scholar : PubMed/NCBI
6	Schwartzbaum JA, Fisher JL, Aldape KD and Wrensch M: Epidemiology and molecular pathology of glioma. Nat Clin Pract Neurol. 2:494–503. 2006.quiz 491 p following 516. View Article : Google Scholar : PubMed/NCBI
7	Gross B, Gaestel M, Böhm H and Bielka H: cDNA sequence coding for a translationally controlled human tumor protein. Nucleic Acids Res. 17:83671989. View Article : Google Scholar : PubMed/NCBI
8	Chung S, Kim M, Choi W, Chung J and Lee K: Expression of translationally controlled tumor protein mRNA in human colon cancer. Cancer Lett. 156:185–190. 2000. View Article : Google Scholar : PubMed/NCBI
9	Bazile F, Pascal A, Arnal I, Le Clainche C, Chesnel F and Kubiak JZ: Complex relationship between TCTP, microtubules and actin microfilaments regulates cell shape in normal and cancer cells. Carcinogenesis. 30:555–565. 2009. View Article : Google Scholar : PubMed/NCBI
10	Zhang F, Liu B, Wang Z, Yu XJ, Ni QX, Yang WT, Mukaida N and Li YY: A novel regulatory mechanism of Pim-3 kinase stability and its involvement in pancreatic cancer progression. Mol Cancer Res. 11:1508–1520. 2013. View Article : Google Scholar : PubMed/NCBI
11	Liu LK, Wu HF, Guo ZR, Chen XJ, Yang D, Shu YQ and Zhang JN: Targeted efficacy of dihydroartemisinin for transla-tionally controlled protein expression in a lung cancer model. Asian Pac J Cancer Prev. 15:2511–2515. 2014. View Article : Google Scholar
12	Cans C, Passer BJ, Shalak V, Nancy-Portebois V, Crible V, Amzallag N, Allanic D, Tufino R, Argentini M, Moras D, et al: Translationally controlled tumor protein acts as a guanine nucleotide dissociation inhibitor on the translation elongation factor eEF1A. Proc Natl Acad Sci USA. 100:13892–13897. 2003. View Article : Google Scholar : PubMed/NCBI
13	Gachet Y, Tournier S, Lee M, Lazaris-Karatzas A, Poulton T and Bommer UA: The growth-related, translationally controlled protein P23 has properties of a tubulin binding protein and associates transiently with microtubules during the cell cycle. J Cell Sci. 112:1257–1271. 1999.PubMed/NCBI
14	Liu H, Peng HW, Cheng YS, Yuan HS and Yang-Yen HF: Stabilization and enhancement of the antiapoptotic activity of mcl-1 by TCTP. Mol Cell Biol. 25:3117–3126. 2005. View Article : Google Scholar : PubMed/NCBI
15	Nagano-Ito M and Ichikawa S: Biological effects of Mammalian translationally controlled tumor protein (TCTP) on cell death, proliferation and tumorigenesis. Biochem Res Int. 2012:2049602012. View Article : Google Scholar
16	Li F, Zhang D and Fujise K: Characterization of fortilin, a novel antiapoptotic protein. J Biol Chem. 276:47542–47549. 2001. View Article : Google Scholar : PubMed/NCBI
17	Miao X, Chen YB, Xu SL, Zhao T, Liu JY, Li YR, Wang J, Zhang J and Guo GZ: TCTP overexpression is associated with the development and progression of glioma. Tumour Biol. 34:3357–3361. 2013. View Article : Google Scholar : PubMed/NCBI
18	Gu X, Yao L, Ma G, Cui L, Li Y, Liang W, Zhao B and Li K: TCTP promotes glioma cell proliferation in vitro and in vivo via enhanced β-catenin/TCF-4 transcription. Neuro Oncol. 16:217–227. 2014. View Article : Google Scholar :
19	Acunzo J, Baylot V, So A and Rocchi P: TCTP as therapeutic target in cancers. Cancer Treat Rev. 40:760–769. 2014. View Article : Google Scholar : PubMed/NCBI
20	Lucibello M, Gambacurta A, Zonfrillo M, Pierimarchi P, Serafino A, Rasi G, Rubartelli A and Garaci E: TCTP is a critical survival factor that protects cancer cells from oxidative stress-induced cell-death. Exp Cell Res. 317:2479–2489. 2011. View Article : Google Scholar : PubMed/NCBI
21	Graidist P, Phongdara A and Fujise K: Antiapoptotic protein partners fortilin and MCL1 independently protect cells from 5-fluorouracil-induced cytotoxicity. J Biol Chem. 279:40868–40875. 2004. View Article : Google Scholar : PubMed/NCBI
22	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
23	Chan TH, Chen L and Guan XY: Role of translationally controlled tumor protein in cancer progression. Biochem Res Int. 2012:3693842012. View Article : Google Scholar : PubMed/NCBI
24	Tuynder M, Fiucci G, Prieur S, Lespagnol A, Géant A, Beaucourt S, Duflaut D, Besse S, Susini L, Cavarelli J, et al: Translationally controlled tumor protein is a target of tumor reversion. Proc Natl Acad Sci USA. 101:15364–15369. 2004. View Article : Google Scholar : PubMed/NCBI
25	Chen SH, Wu PS, Chou CH, Yan YT, Liu H, Weng SY and Yang-Yen HF: A knockout mouse approach reveals that TCTP functions as an essential factor for cell proliferation and survival in a tissue-or cell type-specific manner. Mol Biol Cell. 18:2525–2532. 2007. View Article : Google Scholar : PubMed/NCBI
26	Zhu WL, Cheng HX, Han N, Liu DL, Zhu WX, Fan BL and Duan FL: Messenger RNA expression of translationally controlled tumor protein (TCTP) in liver regeneration and cancer. Anticancer Res. 28:1575–1580. 2008.PubMed/NCBI
27	Hsiao YC, Hsieh YS, Kuo WH, Chiou HL, Yang SF, Chiang WL and Chu SC: The tumor-growth inhibitory activity of flavanone and 2′-OH flavanone in vitro and in vivo through induction of cell cycle arrest and suppression of cyclins and CDKs. J Biomed Sci. 14:107–119. 2007. View Article : Google Scholar
28	Ayyagari VN and Brard L: Bithionol inhibits ovarian cancer cell growth in vitro-studies on mechanism(s) of action. BMC Cancer. 14:612014. View Article : Google Scholar
29	He L, Lu N, Dai Q, Zhao Y, Zhao L, Wang H, Li Z, You Q and Guo Q: Wogonin induced G1 cell cycle arrest by regulating Wnt/β-catenin signaling pathway and inactivating CDK8 in human colorectal cancer carcinoma cells. Toxicology. 312:36–47. 2013. View Article : Google Scholar : PubMed/NCBI
30	Jung J, Kim HY, Maeng J, Kim M, Shin DH and Lee K: Interaction of translationally controlled tumor protein with Apaf-1 is involved in the development of chemoresistance in HeLa cells. BMC Cancer. 14:1652014. View Article : Google Scholar : PubMed/NCBI
31	Susini L, Besse S, Duflaut D, Lespagnol A, Beekman C, Fiucci G, Atkinson AR, Busso D, Poussin P, Marine JC, et al: TCTP protects from apoptotic cell death by antagonizing bax function. Cell Death Differ. 15:1211–1220. 2008. View Article : Google Scholar : PubMed/NCBI
32	Wu D, Guo Z, Min W, Zhou B, Li M, Li W and Luo D: Upregulation of TCTP expression in human skin squamous cell carcinoma increases tumor cell viability through anti-apoptotic action of the protein. Exp Ther Med. 3:437–442. 2012.PubMed/NCBI
33	Rho SB, Lee JH, Park MS, Byun HJ, Kang S, Seo SS, Kim JY and Park SY: Anti-apoptotic protein TCTP controls the stability of the tumor suppressor p53. FEBS Lett. 585:29–35. 2011. View Article : Google Scholar
34	Kaarbo M, Storm ML, Qu S, Wæhre H, Risberg B, Danielsen HE and Saatcioglu F: TCTP is an androgen-regulated gene implicated in prostate cancer. PLoS One. 8:e693982013. View Article : Google Scholar : PubMed/NCBI
35	Nakopoulou L, Tsirmpa I, Alexandrou P, Louvrou A, Ampela C, Markaki S and Davaris PS: MMP-2 protein in invasive breast cancer and the impact of MMP-2/TIMP-2 phenotype on overall survival. Breast Cancer Res Treat. 77:145–155. 2003. View Article : Google Scholar : PubMed/NCBI
36	Luo Y, Liang F and Zhang ZY: PRL1 promotes cell migration and invasion by increasing MMP2 and MMP9 expression through Src and ERK1/2 pathways. Biochemistry. 48:1838–1846. 2009. View Article : Google Scholar : PubMed/NCBI
37	Chu ZH, Liu L, Zheng CX, Lai W, Li SF, Wu H, Zeng YJ, Zhao HY and Guan YF: Proteomic analysis identifies translationally controlled tumor protein as a mediator of phosphatase of regenerating liver-3-promoted proliferation, migration and invasion in human colon cancer cells. Chin Med J (Engl). 124:3778–3785. 2011.
38	Ma Q, Geng Y, Xu W, Wu Y, He F, Shu W, Huang M, Du H and Li M: The role of translationally controlled tumor protein in tumor growth and metastasis of colon adenocarcinoma cells. J Proteome Res. 9:40–49. 2010. View Article : Google Scholar