Overexpression of human telomerase reverse transcriptase promotes the motility and invasiveness of HepG2 cells in vitro

Chen,Peng-Cheng; Peng,Ji-Run; Huang,Lei; Li,Wen-Xia; Wang,Wen-Zhen; Cui,Zhu-Qing-Qing; Han,Hui; Gong,Lei; Xiang,Da-Peng; Qiao,Shi-Shi; Yu,Xin; Wei,Yu-Hua; Ma,Li-Ping; Li,Na; Zhu,Ji-Ye; Leng,Xi-Sheng

doi:10.3892/or.2013.2563

Overexpression of human telomerase reverse transcriptase promotes the motility and invasiveness of HepG2 cells in vitro

Authors:
- Peng-Cheng Chen
- Ji-Run Peng
- Lei Huang
- Wen-Xia Li
- Wen-Zhen Wang
- Zhu-Qing-Qing Cui
- Hui Han
- Lei Gong
- Da-Peng Xiang
- Shi-Shi Qiao
- Xin Yu
- Yu-Hua Wei
- Li-Ping Ma
- Na Li
- Ji-Ye Zhu
- Xi-Sheng Leng
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Affiliations: Peking University People's Hospital, Beijing 100044, P.R. China
Published online on: June 25, 2013 https://doi.org/10.3892/or.2013.2563
Pages: 1157-1164

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Abstract

Recent studies have indicated that telomerase activity promotes cancer invasion and metastasis, but the underlying mechanism remains unclear. Several studies have shown that expression of exogenous human telomerase reverse transcriptase (hTERT) can promote motility and invasiveness among telomerase-negative tumor cells, and inhibition of endogenous telomerase activity can reduce invasiveness in tumor cells. However, whether overexpression of hTERT can further enhance the motility and invasiveness of telomerase‑positive tumor cells has yet to be determined. In the present study, we showed that stable overexpression of hTERT can increase telomerase activity and telomere length, which significantly promotes the invasive and metastatic potential of telomerase‑positive HepG2 cells but does not affect cell proliferation. Further analysis suggested that enhanced invasiveness and metastasis may act through corresponding upregulation of mRNA and protein expression of matrix metalloproteinase 9 (MMP9) and Ras homolog gene family member C (RhoC). Our study indicated that exogenous expression of hTERT may promote invasiveness and metastasis through upregulation of MMP9 and RhoC.

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1	Morin GB: The human telomere terminal transferase enzyme is a ribonucleoprotein that synthesizes TTAGGG repeats. Cell. 59:521–529. 1989. View Article : Google Scholar : PubMed/NCBI
2	Greider CW and Blackburn EH: The telomere terminal transferase of Tetrahymena is a ribonucleoprotein enzyme with two kinds of primer specificity. Cell. 51:887–898. 1987. View Article : Google Scholar : PubMed/NCBI
3	Greider CW and Blackburn EH: Identification of a specific telomere terminal transferase activity in Tetrahymena extracts. Cell. 43:405–413. 1985. View Article : Google Scholar : PubMed/NCBI
4	Feng J, Funk WD, Wang SS, et al: The RNA component of human telomerase. Science. 269:1236–1241. 1995. View Article : Google Scholar : PubMed/NCBI
5	Toshikuni N, Nouso K, Higashi T, et al: Expression of telomerase-associated protein 1 and telomerase reverse transcriptase in hepatocellular carcinoma. Br J Cancer. 82:833–837. 2000. View Article : Google Scholar : PubMed/NCBI
6	Nakayama J, Tahara H, Tahara E, et al: Telomerase activation by hTRT in human normal fibroblasts and hepatocellular carcinomas. Nat Genet. 18:65–68. 1998. View Article : Google Scholar : PubMed/NCBI
7	Ohta K, Kanamaru T, Morita Y, Hayashi Y, Ito H and Yamamoto M: Telomerase activity in hepatocellular carcinoma as a predictor of postoperative recurrence. J Gastroenterol. 32:791–796. 1997. View Article : Google Scholar : PubMed/NCBI
8	Nakashio R, Kitamoto M, Tahara H, Nakanishi T, Ide T and Kajiyama G: Significance of telomerase activity in the diagnosis of small differentiated hepatocellular carcinoma. Int J Cancer. 74:141–147. 1997. View Article : Google Scholar : PubMed/NCBI
9	Kojima H, Yokosuka O, Imazeki F, Saisho H and Omata M: Telomerase activity and telomere length in hepatocellular carcinoma and chronic liver disease. Gastroenterology. 112:493–500. 1997. View Article : Google Scholar : PubMed/NCBI
10	Ohta K, Kanamaru T, Yamamoto M and Saitoh Y: Clinical significance of telomerase activity in hepatocellular carcinoma. Kobe J Med Sci. 42:207–217. 1996.PubMed/NCBI
11	Ide T, Tahara H, Nakashio R, Kitamoto M, Nakanishi T and Kajiyama G: Telomerase in hepatocellular carcinogenesis. Hum Cell. 9:283–286. 1996.
12	Taga S, Osaki T, Ohgami A, Imoto H and Yasumoto K: Prognostic impact of telomerase activity in non-small cell lung cancers. Ann Surg. 230:715–720. 1999. View Article : Google Scholar : PubMed/NCBI
13	Marchetti A, Bertacca G, Buttitta F, et al: Telomerase activity as a prognostic indicator in stage I non-small cell lung cancer. Clin Cancer Res. 5:2077–2081. 1999.PubMed/NCBI
14	Dome JS, Chung S, Bergemann T, et al: High telomerase reverse transcriptase (hTERT) messenger RNA level correlates with tumor recurrence in patients with favorable histology Wilms’ tumor. Cancer Res. 59:4301–4307. 1999.PubMed/NCBI
15	Okusa Y, Shinomiya N, Ichikura T and Mochizuki H: Correlation between telomerase activity and DNA ploidy in gastric cancer. Oncology. 55:258–264. 1998. View Article : Google Scholar : PubMed/NCBI
16	Bechter OE, Eisterer W, Pall G, Hilbe W, Kühr T and Thaler J: Telomere length and telomerase activity predict survival in patients with B cell chronic lymphocytic leukemia. Cancer Res. 58:4918–4922. 1998.PubMed/NCBI
17	Langford LA, Piatyszek MA, Xu R, Schold SC Jr, Wright WE and Shay JW: Telomerase activity in ordinary meningiomas predicts poor outcome. Hum Pathol. 28:416–420. 1997. View Article : Google Scholar : PubMed/NCBI
18	Hiyama E, Hiyama K, Ohtsu K, et al: Telomerase activity in neuroblastoma: is it a prognostic indicator of clinical behaviour? Eur J Cancer. 33:1932–1936. 1997. View Article : Google Scholar : PubMed/NCBI
19	Clark GM, Osborne CK, Levitt D, Wu F and Kim NW: Telomerase activity and survival of patients with node-positive breast cancer. J Natl Cancer Inst. 89:1874–1881. 1997. View Article : Google Scholar : PubMed/NCBI
20	Hiyama E, Yokoyama T, Tatsumoto N, et al: Telomerase activity in gastric cancer. Cancer Res. 55:3258–3262. 1995.PubMed/NCBI
21	Hiyama E, Hiyama K, Yokoyama T, Matsuura Y, Piatyszek MA and Shay JW: Correlating telomerase activity levels with human neuroblastoma outcomes. Nat Med. 1:249–255. 1995. View Article : Google Scholar : PubMed/NCBI
22	Saini N, Srinivasan R, Chawla Y, Sharma S, Chakraborti A and Rajwanshi A: Telomerase activity, telomere length and human telomerase reverse transcriptase expression in hepatocellular carcinoma is independent of hepatitis virus status. Liver Int. 29:1162–1170. 2009. View Article : Google Scholar
23	Suda T, Isokawa O, Aoyagi Y, et al: Quantitation of telomerase activity in hepatocellular carcinoma: a possible aid for a prediction of recurrent diseases in the remnant liver. Hepatology. 27:402–406. 1998. View Article : Google Scholar : PubMed/NCBI
24	Tatsuma T, Goto S, Kitano S, Lin YC, Lee CM and Chen CL: Telomerase activity in peripheral blood for diagnosis of hepatoma. J Gastroenterol Hepatol. 15:1064–1070. 2000. View Article : Google Scholar : PubMed/NCBI
25	Kobayashi T, Kubota K, Takayama T and Makuuchi M: Telomerase activity as a predictive marker for recurrence of hepatocellular carcinoma after hepatectomy. Am J Surg. 181:284–288. 2001. View Article : Google Scholar : PubMed/NCBI
26	Sun B, Huang Q, Liu S, et al: Progressive loss of malignant behavior in telomerase-negative tumorigenic adrenocortical cells and restoration of tumorigenicity by human telomerase reverse transcriptase. Cancer Res. 64:6144–6151. 2004. View Article : Google Scholar
27	Brachner A, Sasgary S, Pirker C, et al: Telomerase- and alternative telomere lengthening-independent telomere stabilization in a metastasis-derived human non-small cell lung cancer cell line: effect of ectopic hTERT. Cancer Res. 66:3584–3592. 2006. View Article : Google Scholar
28	Yu ST, Chen L, Wang HJ, Tang XD, Fang DC and Yang SM: hTERT promotes the invasion of telomerase-negative tumor cells in vitro. Int J Oncol. 35:329–336. 2009.PubMed/NCBI
29	Yao X, Wang X, Zhang S, Yan L and Zhu H: Inhibitory effect of silencing hTERT gene on growth of human squamous cell carcinoma xenograft in nude mice. Lin Chung Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 25:939–943. 2011.(In Chinese).
30	George J, Banik NL and Ray SK: Knockdown of hTERT and concurrent treatment with interferon-gamma inhibited proliferation and invasion of human glioblastoma cell lines. Int J Biochem Cell Biol. 42:1164–1173. 2010. View Article : Google Scholar : PubMed/NCBI
31	Sato N, Maehara N, Mizumoto K, et al: Telomerase activity of cultured human pancreatic carcinoma cell lines correlates with their potential for migration and invasion. Cancer. 91:496–504. 2001. View Article : Google Scholar : PubMed/NCBI
32	Bagheri S, Nosrati M, Li S, et al: Genes and pathways downstream of telomerase in melanoma metastasis. Proc Natl Acad Sci USA. 103:11306–11311. 2006. View Article : Google Scholar : PubMed/NCBI
33	Nart D, Yaman B, Yilmaz F, Zeytunlu M, Karasu Z and Kiliç M: Expression of matrix metalloproteinase-9 in predicting prognosis of hepatocellular carcinoma after liver transplantation. Liver Transpl. 16:621–630. 2010.PubMed/NCBI
34	Arii S, Mise M, Harada T, et al: Overexpression of matrix metalloproteinase 9 gene in hepatocellular carcinoma with invasive potential. Hepatology. 24:316–322. 1996. View Article : Google Scholar : PubMed/NCBI
35	Clark EA, Golub TR, Lander ES and Hynes RO: Genomic analysis of metastasis reveals an essential role for RhoC. Nature. 406:532–535. 2000. View Article : Google Scholar : PubMed/NCBI
36	Ridley AJ: Rho GTPases and cell migration. J Cell Sci. 114:2713–2722. 2001.PubMed/NCBI
37	Van Aelst L and D’Souza-Schorey C: Rho GTPases and signaling networks. Genes Dev. 11:2295–2322. 1997.PubMed/NCBI
38	Wang W, Yang LY, Huang GW, et al: Genomic analysis reveals RhoC as a potential marker in hepatocellular carcinoma with poor prognosis. Br J Cancer. 90:2349–2355. 2004.PubMed/NCBI
39	Wang W, Yang LY, Yang ZL, Huang GW and Lu WQ: Expression and significance of RhoC gene in hepatocellular carcinoma. World J Gastroenterol. 9:1950–1953. 2003.PubMed/NCBI
40	Okabe H, Satoh S, Kato T, et al: Genome-wide analysis of gene expression in human hepatocellular carcinomas using cDNA microarray: identification of genes involved in viral carcinogenesis and tumor progression. Cancer Res. 61:2129–2137. 2001.
41	Kleer CG, Griffith KA, Sabel MS, et al: RhoC-GTPase is a novel tissue biomarker associated with biologically aggressive carcinomas of the breast. Breast Cancer Res Treat. 93:101–110. 2005. View Article : Google Scholar : PubMed/NCBI
42	Shikada Y, Yoshino I, Okamoto T, Fukuyama S, Kameyama T and Maehara Y: Higher expression of RhoC is related to invasiveness in non-small cell lung carcinoma. Clin Cancer Res. 9:5282–5286. 2003.PubMed/NCBI
43	Xue F, Takahara T, Yata Y, et al: Blockade of Rho/Rho-associated coiled coil-forming kinase signaling can prevent progression of hepatocellular carcinoma in matrix metalloproteinase-dependent manner. Hepatol Res. 38:810–817. 2008. View Article : Google Scholar
44	Iiizumi M, Bandyopadhyay S, Pai SK, et al: RhoC promotes metastasis via activation of the Pyk2 pathway in prostate cancer. Cancer Res. 68:7613–7620. 2008. View Article : Google Scholar : PubMed/NCBI
45	Belair CD, Yeager TR, Lopez PM and Reznikoff CA: Telomerase activity: a biomarker of cell proliferation, not malignant transformation. Proc Natl Acad Sci USA. 94:13677–13682. 1997. View Article : Google Scholar : PubMed/NCBI
46	Smith LL, Coller HA and Roberts JM: Telomerase modulates expression of growth-controlling genes and enhances cell proliferation. Nat Cell Biol. 5:474–479. 2003. View Article : Google Scholar : PubMed/NCBI
47	Gronthos S, Chen S, Wang CY, Robey PG and Shi S: Telomerase accelerates osteogenesis of bone marrow stromal stem cells by upregulation of CBFA1, osterix, and osteocalcin. J Bone Miner Res. 18:716–722. 2003. View Article : Google Scholar : PubMed/NCBI
48	Xiang H, Wang J, Mao Y, Liu M, Reddy VN and Li DW: Human telomerase accelerates growth of lens epithelial cells through regulation of the genes mediating RB/E2F pathway. Oncogene. 21:3784–3791. 2002. View Article : Google Scholar : PubMed/NCBI
49	Bednarek AK, Sahin A, Brenner AJ, Johnston DA and Aldaz CM: Analysis of telomerase activity levels in breast cancer: positive detection at the in situ breast carcinoma stage. Clin Cancer Res. 3:11–16. 1997.PubMed/NCBI