Oxidative stress may enhance the malignant potential of human hepatocellular carcinoma by telomerase activation
Taichiro Nishikawa
Kyoto Prefectural University of Medicine Graduate School of Medical Science, Molecular Gastroenterology and Hepatology, Kyoto, Japan
Search for more papers by this authorTomoki Nakajima
Kyoto Prefectural University of Medicine Graduate School of Medical Science, Molecular Gastroenterology and Hepatology, Kyoto, Japan
Search for more papers by this authorTatsuo Katagishi
Kyoto Prefectural University of Medicine Graduate School of Medical Science, Molecular Gastroenterology and Hepatology, Kyoto, Japan
Search for more papers by this authorYoshihisa Okada
Kyoto Prefectural University of Medicine Graduate School of Medical Science, Molecular Gastroenterology and Hepatology, Kyoto, Japan
Search for more papers by this authorMasayasu Jo
Kyoto Prefectural University of Medicine Graduate School of Medical Science, Molecular Gastroenterology and Hepatology, Kyoto, Japan
Search for more papers by this authorKeizo Kagawa
Kyoto Prefectural University of Medicine Graduate School of Medical Science, Molecular Gastroenterology and Hepatology, Kyoto, Japan
Search for more papers by this authorTakeshi Okanoue
Kyoto Prefectural University of Medicine Graduate School of Medical Science, Molecular Gastroenterology and Hepatology, Kyoto, Japan
Search for more papers by this authorYoshito Itoh
Kyoto Prefectural University of Medicine Graduate School of Medical Science, Molecular Gastroenterology and Hepatology, Kyoto, Japan
Search for more papers by this authorToshikazu Yoshikawa
Kyoto Prefectural University of Medicine Graduate School of Medical Science, Molecular Gastroenterology and Hepatology, Kyoto, Japan
Search for more papers by this authorTaichiro Nishikawa
Kyoto Prefectural University of Medicine Graduate School of Medical Science, Molecular Gastroenterology and Hepatology, Kyoto, Japan
Search for more papers by this authorTomoki Nakajima
Kyoto Prefectural University of Medicine Graduate School of Medical Science, Molecular Gastroenterology and Hepatology, Kyoto, Japan
Search for more papers by this authorTatsuo Katagishi
Kyoto Prefectural University of Medicine Graduate School of Medical Science, Molecular Gastroenterology and Hepatology, Kyoto, Japan
Search for more papers by this authorYoshihisa Okada
Kyoto Prefectural University of Medicine Graduate School of Medical Science, Molecular Gastroenterology and Hepatology, Kyoto, Japan
Search for more papers by this authorMasayasu Jo
Kyoto Prefectural University of Medicine Graduate School of Medical Science, Molecular Gastroenterology and Hepatology, Kyoto, Japan
Search for more papers by this authorKeizo Kagawa
Kyoto Prefectural University of Medicine Graduate School of Medical Science, Molecular Gastroenterology and Hepatology, Kyoto, Japan
Search for more papers by this authorTakeshi Okanoue
Kyoto Prefectural University of Medicine Graduate School of Medical Science, Molecular Gastroenterology and Hepatology, Kyoto, Japan
Search for more papers by this authorYoshito Itoh
Kyoto Prefectural University of Medicine Graduate School of Medical Science, Molecular Gastroenterology and Hepatology, Kyoto, Japan
Search for more papers by this authorToshikazu Yoshikawa
Kyoto Prefectural University of Medicine Graduate School of Medical Science, Molecular Gastroenterology and Hepatology, Kyoto, Japan
Search for more papers by this authorAbstract
Background/Aims: Continuous oxidative stress (OS) plays an important role in the progression of chronic liver diseases and hepatocarcinogenesis through telomere shortening in hepatocytes. However, it has not been established how the OS influences the progression of human hepatocellular carcinomas (HCCs). We examined the correlations of OS with telomere length of cancer cells, telomerase activity and other clinicopathological factors in 68 HCCs.
Methods: The level of 8-hydroxy-2′-deoxyguanosine (8-OHdG) as a marker of OS was examined immunohistochemically and OS was scored in four grades (0–3). The telomere length of cancer cells was measured by quantitative fluorescence in situ hybridization. Telomerase activity was measured by (i) immunodetection of human telomerase reverse transcriptase (hTERT) and (ii) telomere repeat amplification protocol (TRAP) assay. Telomerase related proteins, phosphatase and tensin homolog deleted on chromosome 10 (PTEN) and Akt, and other clinicopathological factors were also evaluated.
Results: As the OS grade increased, the average telomere length became significantly shorter in HCCs, especially in the hTERT-negative group. In the state of high-grade OS, hTERT-positive HCC cells showed more proliferative and less apoptotic features compared with hTERT-negative HCC cells. Telomerase activity, as measured by the TRAP assay, was strongly correlated with OS grade in HCCs. Furthermore, a high OS grade was correlated with the downexpression of PTEN and the activation of Akt.
Conclusions: Oxidative stress enhanced the malignant potential of HCCs through the activation of telomerase, which raises the possibility of using OS as a marker for assessing the clinical state of HCCs.
References
- 1 Jain SK, Pemberton PW, Smith A, et al. Oxidative stress in chronic hepatitis C: not just a feature of late stage disease. J Hepatol 2002; 36: 805–11.
- 2 Aboutwerat A, Pemberton PW, Smith A, et al. Oxidant stress is a significant feature of primary biliary cirrhosis. Biochim Biophys Acta 2003; 1637: 142–50.
- 3 Jungst C, Cheng B, Gehrke R, et al. Oxidative damage is increased in human liver tissue adjacent to hepatocellular carcinoma. Hepatology 2004; 39: 1663–72.
- 4 Blackburn EH. Structure and function of telomeres. Nature 1991; 350: 569–73.
- 5 Levy MZ, Allsopp RC, Futcher AB, et al. Telomere end-replication problem and cell aging. J Mol Biol 1992; 225: 951–60.
- 6 Blackburn EH. Switching and signaling at the telomere. Cell 2001; 106: 661–73.
- 7 Saretzki G, Von Zglinicki T. Replicative aging, telomeres, and oxidative stress. Ann NY Acad Sci 2002; 959: 24–9.
- 8 Kurz DJ, Decary S, Hong Y, et al. Chronic oxidative stress compromises telomere integrity and accelerates the onset of senescence in human endothelial cells. J Cell Sci 2004; 117: 2417–26.
- 9 Wiemann SU, Satyanarayana A, Tsahuridu M, et al. Hepatocyte telomere shortening and senescence are general markers of human liver cirrhosis. FASEB J 2002; 16: 935–42.
- 10 Aikata H, Takaishi H, Kawakami Y, et al. Telomere reduction in human liver tissues with age and chronic inflammation. Exp Cell Res 2000; 256: 578–82.
- 11 Oh BK, Jo Chae K, Park C, et al. Telomere shortening and telomerase reactivation in dysplastic nodules of human hepatocarcinogenesis. J Hepatol 2003; 39: 786–92.
- 12 Desmet VJ, Gerber M, Hoofnargle JH, et al. Classification of chronic hepatitis: diagnosis, grading and staging. Hepatology 1994; 19: 1513–20.
- 13 Kato J, Kobune M, Nakamura T, et al. Normalization of elevated hepatic 8-hydroxy-2′-deoxyguanosine levels in chronic hepatitis C patients by phlebotomy and low iron diet. Cancer Res 2001; 61: 8697–702.
- 14 Hisatomi H, Nagao K, Kanamaru T, et al. Levels of telomerase catalytic subunit mRNA as a predictor of potential malignancy. Int J Oncol 1999; 14: 727–32.
- 15 Kawakami Y, Kitamoto M, Nakanishi T, et a l. Immuno-histochemical detection of human telomerase reverse transcriptase in human liver tissues. Oncogene 2000; 19: 3888–93.
- 16 Moriguchi M, Nakajima T, Kimura H, et al. The copper chelator trientine has an antiangiogenic effect against hepatocellular carcinoma, possibly through inhibition of interleukin-8 production. Int J Cancer 2002; 102: 445–52.
- 17 Nakajima T, Katagishi T, Moriguchi M, et al. Tumor size-independence of telomere length indicates an aggressive feature of HCC. Biochem Biophys Res Commun 2004; 325: 1131–5.
- 18 Meeker AK, Gage WR, Hicks JL, et al. Telomere length assessment in human archival tissues: combined telomere fluorescence in situ hybridization and immunostaining. Am J Pathol 2002; 160: 1259–68.
- 19 Iwama H, Ohyashiki K, Ohyashiki JH, et al. Telomeric length and telomerase activity vary with age in peripheral blood cells obtained from normal individuals. Hum Genet 1998; 102: 397–402.
- 20 Kang SS, Kwon T, Kwon DY, Do SI. Akt protein kinase enhances human telomerase activity through phosphorylation of telomerase reverse transcriptase subunit. J Biol Chem 1999; 274: 13085–90.
- 21 Kimura A, Ohmichi M, Kawagoe J, et al. Induction of hTERT expression and phosphorylation by estrogen via Akt cascade in human ovarian cancer cell lines. Oncogene 2004; 23: 4505–15.
- 22 Haas-Kogan D, Shalev N, Wong M, et al. Protein kinase B (PKB/Akt) activity is elevated in glioblastoma cells due to mutation of the tumor suppressor PTEN/MMAC. Curr Biol 1998; 8: 1195–8.
- 23 Wu SK, Wang BJ, Yang Y, et al. Expression of PTEN, PPM1A and P-Smad2 in hepatocellular carcinomas and adjacent liver tissues. World J Gastroenterol 2007; 13: 4554–9.
- 24 Seki S, Kitada T, Yamada T, et al. In situ detection of lipid peroxidation and oxidative DNA damage in non-alcoholic fatty liver diseases. J Hepatol 2002; 37: 56–62.
- 25 Ichiba M, Maeta Y, Mukoyama T, et al. Expression of 8-hydroxy-2′-deoxyguanosine in chronic liver disease and hepatocellular carcinoma. Liver Int 2003; 23: 338–45.
- 26 Szatrowski TP, Nathan CF. Production of large amounts of hydrogen peroxide by human tumor cells. Cancer Res 1991; 51: 794–8.
- 27 Toyokuni S, Okamoto K, Yodoi J, Hiai H. Persistent oxidative stress in cancer. FEBS Lett 1995; 358: 1–3.
- 28 Chandel NS, McClintock DS, Feliciano CE, et al. Reactive oxygen species generated at mitochondrial complex III stabilize hypoxia-inducible factor-1alpha during hypoxia: a mechanism of O2 sensing. J Biol Chem 2000; 275: 25130–8.
- 29 Maulik N. Redox signaling of angiogenesis. Antioxid Redox Signal 2002; 4: 805–15.
- 30 Sekoguchi S, Nakajima T, Moriguchi M, et al. Role of cell-cycle turnover and oxidative stress in telomere shortening and cellular senescence in patients with chronic hepatitis C. J Gastroenterol Hepatol 2007; 22: 182–90.
- 31 Nakajima T, Moriguchi M, Katagishi T, et al. Premature telomere shortening and impaired regenerative response in hepatocytes of individuals with NAFLD. Liver Int 2006; 26: 23–31.
- 32 Nagao K, Tomimatsu M, Endo H, et al. Telomerase reverse transcriptase mRNA expression and telomerase activity in hepatocellular carcinoma. J Gastroenterol 1999; 34: 83–7.
- 33 Miura N, Shiota G, Nakagawa T, et al. Sensitive detection of human telomerase reverse transcriptase mRNA in the serum of patients with hepatocellular carcinoma. Oncology 2003; 64: 430–4.
- 34 Kraemer K, Fuessel S, Schmidt U, et al. Antisense-mediated hTERT inhibition specifically reduces the growth of human bladder cancer cells. Clin Cancer Res 2003; 9: 3794–800.
- 35 Cao Y, Li H, Deb S, Liu JP. TERT regulates cell survival independent of telomerase enzymatic activity. Oncogene 2002; 21: 3130–8.
- 36 Liu DY, Peng ZH, Qiu GQ, Zhou CZ. Expression of telomerase activity and oxidative stress in human hepatocellular carcinoma with cirrhosis. World J Gastroenterol 2003; 9: 1859–62.
- 37 Liu K, Hodes RJ, Weng NP. Cutting edge: telomerase activation in human T lymphocytes does not require increase in telomerase reverse transcriptase (hTERT) protein but is associated with hTERT phosphorylation and nuclear translocation. J Immunol 2001; 166: 4826–30.
- 38 Dong-Yun S, Yu-Ru D, Shan-Lin L, et al. Redox stress regulates cell proliferation and apoptosis of human hepatoma through Akt protein phosphorylation. FEBS Lett 2003; 542: 60–4.
- 39 Leslie NR, Bennett D, Lindsay YE, et al. Redox regulation of PI 3-kinase signalling via inactivation of PTEN. EMBO J 2003; 22: 5501–10.
- 40 Shiota G, Maeta Y, Mukoyama T, et al. Effects of Sho-Saiko-to on hepatocarcinogenesis and 8-hydroxy-2′-deoxyguanosine formation. Hepatology 2002; 35: 1125–33.
- 41 Wada S, Satomi Y, Murakoshi M, et al. Tumor suppressive effects of tocotrienol in vivo and in vitro. Cancer Lett 2005; 229: 181–91.
- 42 Nishikawa T, Nakajima T, Moriguchi M, et al. A green tea polyphenol, epigalocatechin-3-gallate, induces apoptosis of human hepatocellular carcinoma, possibly through inhibition of Bcl-2 family proteins. J Hepatol 2006; 44: 1074–82.
- 43 Naasani I, Oh-Hashi F, Oh-Hara T, et al. Blocking telomerase by dietary polyphenols is a major mechanism for limiting the growth of human cancer cells in vitro and in vivo. Cancer Res 2003; 63: 824–30.
- 44 Tauchi T, Nakajima A, Sashida G, et al. Inhibition of human telomerase enhances the effect of the tyrosine kinase inhibitor, imatinib, in BCR–ABL-positive leukemia cells. Clin Cancer Res 2002; 8: 3341–7.
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