Hepatitis C Virus Core Protein Down-Regulates p21Waf1/Cip1 and Inhibits Curcumin-Induced Apoptosis through MicroRNA-345 Targeting in Human Hepatoma Cells

Tzu-Yue Shiu, Shih-Ming Huang, Yu-Lueng Shih, Heng-Cheng Chu, Wei-Kuo Chang, Tsai-Yuan Hsieh

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Background: Hepatitis C virus (HCV) has been reported to regulate cellular microRNAs. The HCV core protein is considered to be a potential oncoprotein in HCV-related hepatocellular carcinoma, but HCV core-modulated cellular microRNAs are unknown. The HCV core protein regulates p21Waf1/Cip1 expression. However, the mechanism of HCV core-associated p21Waf1/Cip1 regulation remains to be further clarified. Therefore, we attempted to determine whether HCV core-modulated cellular microRNAs play an important role in regulating p21Waf1/Cip1 expression in human hepatoma cells. Methods: Cellular microRNA profiling was investigated in core-overexpressing hepatoma cells using TaqMan low density array. Array data were further confirmed by TaqMan real-time qPCR for single microRNA in core-overexpressing and full-length HCV replicon-expressing cells. The target gene of microRNA was examined by reporter assay. The gene expression was determined by real-time qPCR and Western blotting. Apoptosis was examined by annexin V-FITC apoptosis assay. Cell cycle analysis was performed by propidium iodide staining. Cell proliferation was analyzed by MTT assay. Results: HCV core protein up- or down-regulated some cellular microRNAs in Huh7 cells. HCV core-induced microRNA-345 suppressed p21Waf1/Cip1 gene expression through targeting its 3′ untranslated region in human hepatoma cells. Moreover, the core protein inhibited curcumin-induced apoptosis through p21Waf1/Cip1-targeting microRNA-345 in Huh7 cells. Conclusion and Significance: HCV core protein enhances the expression of microRNA-345 which then down-regulates p21Waf1/Cip1 expression. It is the first time that HCV core protein has ever been shown to suppress p21Waf1/Cip1 gene expression through miR-345 targeting. © 2013 Shiu et al.
Original languageEnglish
JournalPLoS One
Volume8
Issue number4
DOIs
Publication statusPublished - 2013
Externally publishedYes

Fingerprint

Curcumin
curcumin
Hepatitis C virus
hepatoma
MicroRNAs
microRNA
Hepatocellular Carcinoma
Down-Regulation
apoptosis
Apoptosis
Viruses
Hepacivirus
proteins
cells
Gene expression
Assays
Gene Expression
gene expression
Hepatitis C virus nucleocapsid protein
assays

Keywords

  • core protein
  • curcumin
  • cyclin dependent kinase inhibitor 1
  • microRNA
  • microRNA 345
  • unclassified drug
  • 3' untranslated region
  • apoptosis
  • article
  • cell cycle
  • cell proliferation
  • cell strain
  • cell strain Huh7
  • controlled study
  • down regulation
  • gene overexpression
  • gene targeting
  • Hepatitis C virus
  • hepatoma cell
  • human
  • human cell
  • nonhuman
  • p21Waf1Cip1 gene
  • protein expression
  • replicon
  • upregulation

Cite this

Hepatitis C Virus Core Protein Down-Regulates p21Waf1/Cip1 and Inhibits Curcumin-Induced Apoptosis through MicroRNA-345 Targeting in Human Hepatoma Cells. / Shiu, Tzu-Yue; Huang, Shih-Ming; Shih, Yu-Lueng; Chu, Heng-Cheng; Chang, Wei-Kuo; Hsieh, Tsai-Yuan.

In: PLoS One, Vol. 8, No. 4, 2013.

Research output: Contribution to journalArticle

@article{170cce6fd07a4e3fa4ca780b26c80ddf,
title = "Hepatitis C Virus Core Protein Down-Regulates p21Waf1/Cip1 and Inhibits Curcumin-Induced Apoptosis through MicroRNA-345 Targeting in Human Hepatoma Cells",
abstract = "Background: Hepatitis C virus (HCV) has been reported to regulate cellular microRNAs. The HCV core protein is considered to be a potential oncoprotein in HCV-related hepatocellular carcinoma, but HCV core-modulated cellular microRNAs are unknown. The HCV core protein regulates p21Waf1/Cip1 expression. However, the mechanism of HCV core-associated p21Waf1/Cip1 regulation remains to be further clarified. Therefore, we attempted to determine whether HCV core-modulated cellular microRNAs play an important role in regulating p21Waf1/Cip1 expression in human hepatoma cells. Methods: Cellular microRNA profiling was investigated in core-overexpressing hepatoma cells using TaqMan low density array. Array data were further confirmed by TaqMan real-time qPCR for single microRNA in core-overexpressing and full-length HCV replicon-expressing cells. The target gene of microRNA was examined by reporter assay. The gene expression was determined by real-time qPCR and Western blotting. Apoptosis was examined by annexin V-FITC apoptosis assay. Cell cycle analysis was performed by propidium iodide staining. Cell proliferation was analyzed by MTT assay. Results: HCV core protein up- or down-regulated some cellular microRNAs in Huh7 cells. HCV core-induced microRNA-345 suppressed p21Waf1/Cip1 gene expression through targeting its 3′ untranslated region in human hepatoma cells. Moreover, the core protein inhibited curcumin-induced apoptosis through p21Waf1/Cip1-targeting microRNA-345 in Huh7 cells. Conclusion and Significance: HCV core protein enhances the expression of microRNA-345 which then down-regulates p21Waf1/Cip1 expression. It is the first time that HCV core protein has ever been shown to suppress p21Waf1/Cip1 gene expression through miR-345 targeting. {\circledC} 2013 Shiu et al.",
keywords = "core protein, curcumin, cyclin dependent kinase inhibitor 1, microRNA, microRNA 345, unclassified drug, 3' untranslated region, apoptosis, article, cell cycle, cell proliferation, cell strain, cell strain Huh7, controlled study, down regulation, gene overexpression, gene targeting, Hepatitis C virus, hepatoma cell, human, human cell, nonhuman, p21Waf1Cip1 gene, protein expression, replicon, upregulation",
author = "Tzu-Yue Shiu and Shih-Ming Huang and Yu-Lueng Shih and Heng-Cheng Chu and Wei-Kuo Chang and Tsai-Yuan Hsieh",
note = "被引用次數:14 Export Date: 22 March 2016 通訊地址: Hsieh, T.-Y.; Graduate Institute of Medical Sciences, National Defense Medical Center, Taipei, Taiwan; 電子郵件: tyh1216@ms46.hinet.net 化學物質/CAS: curcumin, 458-37-7 參考文獻: Castello, G., Scala, S., Palmieri, G., Curley, S.A., Izzo, F., HCV-related hepatocellular carcinoma: From chronic inflammation to cancer (2010) Clin Immunol, 134, pp. 237-250; Levrero, M., Viral hepatitis and liver cancer: the case of hepatitis C (2006) Oncogene, 25, pp. 3834-3847; Liu, Q., Tackney, C., Bhat, R.A., Prince, A.M., Zhang, P., Regulated processing of hepatitis C virus core protein is linked to subcellular localization (1997) J Virol, 71, pp. 657-662; Weihofen, A., Binns, K., Lemberg, M.K., Ashman, K., Martoglio, B., Identification of signal peptide peptidase, a presenilin-type aspartic protease (2002) Science, 296, pp. 2215-2218; Lo, S.Y., Masiarz, F., Hwang, S.B., Lai, M.M., Ou, J.H., Differential subcellular localization of hepatitis C virus core gene products (1995) Virology, 213, pp. 455-461; Suzuki, R., Tamura, K., Li, J., Ishii, K., Matsuura, Y., Ubiquitin-mediated degradation of hepatitis C virus core protein is regulated by processing at its carboxyl terminus (2001) Virology, 280, pp. 301-309; Jin, D.Y., Wang, H.L., Zhou, Y., Chun, A.C., Kibler, K.V., Hepatitis C virus core protein-induced loss of LZIP function correlates with cellular transformation (2000) EMBO J, 19, pp. 729-740; You, L.R., Chen, C.M., Yeh, T.S., Tsai, T.Y., Mai, R.T., Hepatitis C virus core protein interacts with cellular putative RNA helicase (1999) J Virol, 73, pp. 2841-2853; Yoshida, T., Hanada, T., Tokuhisa, T., Kosai, K., Sata, M., Activation of STAT3 by the hepatitis C virus core protein leads to cellular transformation (2002) J Exp Med, 196, pp. 641-653; Tsutsumi, T., Suzuki, T., Shimoike, T., Suzuki, R., Moriya, K., Interaction of hepatitis C virus core protein with retinoid X receptor alpha modulates its transcriptional activity (2002) Hepatology, 35, pp. 937-946; Moriishi, K., Okabayashi, T., Nakai, K., Moriya, K., Koike, K., Proteasome activator PA28gamma-dependent nuclear retention and degradation of hepatitis C virus core protein (2003) J Virol, 77, pp. 10237-10249; Moriishi, K., Mochizuki, R., Moriya, K., Miyamoto, H., Mori, Y., Critical role of PA28gamma in hepatitis C virus-associated steatogenesis and hepatocarcinogenesis (2007) Proc Natl Acad Sci U S A, 104, pp. 1661-1666; Lewis, B.P., Burge, C.B., Bartel, D.P., Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets (2005) Cell, 120, pp. 15-20; Brodersen, P., Voinnet, O., Revisiting the principles of microRNA target recognition and mode of action (2009) Nat Rev Mol Cell Biol, 10, pp. 141-148; Esquela-Kerscher, A., Slack, F.J., Oncomirs - microRNAs with a role in cancer (2006) Nat Rev Cancer, 6, pp. 259-269; Gaur, A., Jewell, D.A., Liang, Y., Ridzon, D., Moore, J.H., Characterization of microRNA expression levels and their biological correlates in human cancer cell lines (2007) Cancer Res, 67, pp. 2456-2468; Varnholt, H., Drebber, U., Schulze, F., Wedemeyer, I., Schirmacher, P., MicroRNA gene expression profile of hepatitis C virus-associated hepatocellular carcinoma (2008) Hepatology, 47, pp. 1223-1232; Jiang, J., Gusev, Y., Aderca, I., Mettler, T.A., Nagorney, D.M., Association of MicroRNA expression in hepatocellular carcinomas with hepatitis infection, cirrhosis, and patient survival (2008) Clin Cancer Res, 14, pp. 419-427; Braconi, C., Valeri, N., Gasparini, P., Huang, N., Taccioli, C., Hepatitis C virus proteins modulate microRNA expression and chemosensitivity in malignant hepatocytes (2010) Clin Cancer Res, 16, pp. 957-966; Abbas, T., Dutta, A., p21 in cancer: intricate networks and multiple activities (2009) Nat Rev Cancer, 9, pp. 400-414; Koike, K., Hepatitis C virus contributes to hepatocarcinogenesis by modulating metabolic and intracellular signaling pathways (2007) J Gastroenterol Hepatol, 22, pp. S108-S111; Darvesh, A.S., Aggarwal, B.B., Bishayee, A., Curcumin and liver cancer: a review (2012) Curr Pharm Biotechnol, 13, pp. 218-228; Srivastava, R.K., Chen, Q., Siddiqui, I., Sarva, K., Shankar, S., Linkage of curcumin-induced cell cycle arrest and apoptosis by cyclin-dependent kinase inhibitor p21(/WAF1/CIP1) (2007) Cell Cycle, 6, pp. 2953-2961; Wang, W.Z., Cheng, J., Luo, J., Zhuang, S.M., Abrogation of G2/M arrest sensitizes curcumin-resistant hepatoma cells to apoptosis (2008) FEBS Lett, 582, pp. 2689-2695; Moriya, K., Fujie, H., Shintani, Y., Yotsuyanagi, H., Tsutsumi, T., The core protein of hepatitis C virus induces hepatocellular carcinoma in transgenic mice (1998) Nat Med, 4, pp. 1065-1067; Lee, M.N., Jung, E.Y., Kwun, H.J., Jun, H.K., Yu, D.Y., Hepatitis C virus core protein represses the p21 promoter through inhibition of a TGF-beta pathway (2002) J Gen Virol, 83, pp. 2145-2151; Yamanaka, T., Uchida, M., Doi, T., Innate form of HCV core protein plays an important role in the localization and the function of HCV core protein (2002) Biochem Biophys Res Commun, 294, pp. 521-527; Nguyen, H., Mudryj, M., Guadalupe, M., Dandekar, S., Hepatitis C virus core protein expression leads to biphasic regulation of the p21 cdk inhibitor and modulation of hepatocyte cell cycle (2003) Virology, 312, pp. 245-253; Ohkawa, K., Ishida, H., Nakanishi, F., Hosui, A., Ueda, K., Hepatitis C virus core functions as a suppressor of cyclin-dependent kinase-activating kinase and impairs cell cycle progression (2004) J Biol Chem, 279, pp. 11719-11726; Chen, P.J., Lin, M.H., Tai, K.F., Liu, P.C., Lin, C.J., The Taiwanese hepatitis C virus genome: sequence determination and mapping the 5′ termini of viral genomic and antigenomic RNA (1992) Virology, 188, pp. 102-113; Beard, M.R., Abell, G., Honda, M., Carroll, A., Gartland, M., An infectious molecular clone of a Japanese genotype 1b hepatitis C virus (1999) Hepatology, 30, pp. 316-324; Livak, K.J., Schmittgen, T.D., Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method (2001) Methods, 25, pp. 402-408; Wu, S., Huang, S., Ding, J., Zhao, Y., Liang, L., Multiple microRNAs modulate p21Cip1/Waf1 expression by directly targeting its 3′ untranslated region (2010) Oncogene, 29, pp. 2302-2308; Falc{\'o}n, V., Acosta-Rivero, N., Chinea, G., de la Rosa, M.C., Men{\'e}ndez, I., Nuclear localization of nucleocapsid-like particles and HCV core protein in hepatocytes of a chronically HCV-infected patient (2003) Biochem Biophys Res Commun, 310, pp. 54-58; Yamaguchi, R., Momosaki, S., Gao, G., Hsia, C.C., Kojiro, M., Truncated hepatitis C virus core protein encoded in hepatocellular carcinomas (2004) Int J Mol Med, 14, pp. 1097-1100; Chen, W., Zhang, Z., Chen, J., Zhang, J., Zhang, J., HCV core protein interacts with Dicer to antagonize RNA silencing (2008) Virus Res, 133, pp. 250-258; Pedersen, I.M., Cheng, G., Wieland, S., Volinia, S., Croce, C.M., Interferon modulation of cellular microRNAs as an antiviral mechanism (2007) Nature, 449, pp. 919-922; Tang, J.T., Wang, J.L., Du, W., Hong, J., Zhao, S.L., MicroRNA 345, a methylation-sensitive microRNA is involved in cell proliferation and invasion in human colorectal cancer (2011) Carcinogenesis, 32, pp. 1207-1215; Pan, W., Zhu, S., Yuan, M., Cui, H., Wang, L., MicroRNA-21 and microRNA-148a contribute to DNA hypomethylation in lupus CD4+ T cells by directly and indirectly targeting DNA methyltransferase 1 (2010) J Immunol, 184, pp. 6773-6781; Marquez, R.T., Bandyopadhyay, S., Wendlandt, E.B., Keck, K., Hoffer, B.A., Correlation between microRNA expression levels and clinical parameters associated with chronic hepatitis C viral infection in humans (2010) Lab Invest, 90, pp. 1727-1736; Cervigne, N.K., Reis, P.P., Machado, J., Sadikovic, B., Bradley, G., Identification of a microRNA signature associated with progression of leukoplakia to oral carcinoma (2009) Hum Mol Genet, 18, pp. 4818-4829; Guled, M., Lahti, L., Lindholm, P.M., Salmenkivi, K., Bagwan, I., CDKN2A, NF2, and JUN are dysregulated among other genes by miRNAs in malignant mesothelioma -A miRNA microarray analysis (2009) Genes Chromosomes Cancer, 48, pp. 615-623; Wang, W., Peng, B., Wang, D., Ma, X., Jiang, D., Human tumor microRNA signatures derived from large-scale oligonucleotide microarray datasets (2011) Int J Cancer, 129, pp. 1624-1634",
year = "2013",
doi = "10.1371/journal.pone.0061089",
language = "English",
volume = "8",
journal = "PLoS One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "4",

}

TY - JOUR

T1 - Hepatitis C Virus Core Protein Down-Regulates p21Waf1/Cip1 and Inhibits Curcumin-Induced Apoptosis through MicroRNA-345 Targeting in Human Hepatoma Cells

AU - Shiu, Tzu-Yue

AU - Huang, Shih-Ming

AU - Shih, Yu-Lueng

AU - Chu, Heng-Cheng

AU - Chang, Wei-Kuo

AU - Hsieh, Tsai-Yuan

N1 - 被引用次數:14 Export Date: 22 March 2016 通訊地址: Hsieh, T.-Y.; Graduate Institute of Medical Sciences, National Defense Medical Center, Taipei, Taiwan; 電子郵件: tyh1216@ms46.hinet.net 化學物質/CAS: curcumin, 458-37-7 參考文獻: Castello, G., Scala, S., Palmieri, G., Curley, S.A., Izzo, F., HCV-related hepatocellular carcinoma: From chronic inflammation to cancer (2010) Clin Immunol, 134, pp. 237-250; Levrero, M., Viral hepatitis and liver cancer: the case of hepatitis C (2006) Oncogene, 25, pp. 3834-3847; Liu, Q., Tackney, C., Bhat, R.A., Prince, A.M., Zhang, P., Regulated processing of hepatitis C virus core protein is linked to subcellular localization (1997) J Virol, 71, pp. 657-662; Weihofen, A., Binns, K., Lemberg, M.K., Ashman, K., Martoglio, B., Identification of signal peptide peptidase, a presenilin-type aspartic protease (2002) Science, 296, pp. 2215-2218; Lo, S.Y., Masiarz, F., Hwang, S.B., Lai, M.M., Ou, J.H., Differential subcellular localization of hepatitis C virus core gene products (1995) Virology, 213, pp. 455-461; Suzuki, R., Tamura, K., Li, J., Ishii, K., Matsuura, Y., Ubiquitin-mediated degradation of hepatitis C virus core protein is regulated by processing at its carboxyl terminus (2001) Virology, 280, pp. 301-309; Jin, D.Y., Wang, H.L., Zhou, Y., Chun, A.C., Kibler, K.V., Hepatitis C virus core protein-induced loss of LZIP function correlates with cellular transformation (2000) EMBO J, 19, pp. 729-740; You, L.R., Chen, C.M., Yeh, T.S., Tsai, T.Y., Mai, R.T., Hepatitis C virus core protein interacts with cellular putative RNA helicase (1999) J Virol, 73, pp. 2841-2853; Yoshida, T., Hanada, T., Tokuhisa, T., Kosai, K., Sata, M., Activation of STAT3 by the hepatitis C virus core protein leads to cellular transformation (2002) J Exp Med, 196, pp. 641-653; Tsutsumi, T., Suzuki, T., Shimoike, T., Suzuki, R., Moriya, K., Interaction of hepatitis C virus core protein with retinoid X receptor alpha modulates its transcriptional activity (2002) Hepatology, 35, pp. 937-946; Moriishi, K., Okabayashi, T., Nakai, K., Moriya, K., Koike, K., Proteasome activator PA28gamma-dependent nuclear retention and degradation of hepatitis C virus core protein (2003) J Virol, 77, pp. 10237-10249; Moriishi, K., Mochizuki, R., Moriya, K., Miyamoto, H., Mori, Y., Critical role of PA28gamma in hepatitis C virus-associated steatogenesis and hepatocarcinogenesis (2007) Proc Natl Acad Sci U S A, 104, pp. 1661-1666; Lewis, B.P., Burge, C.B., Bartel, D.P., Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets (2005) Cell, 120, pp. 15-20; Brodersen, P., Voinnet, O., Revisiting the principles of microRNA target recognition and mode of action (2009) Nat Rev Mol Cell Biol, 10, pp. 141-148; Esquela-Kerscher, A., Slack, F.J., Oncomirs - microRNAs with a role in cancer (2006) Nat Rev Cancer, 6, pp. 259-269; Gaur, A., Jewell, D.A., Liang, Y., Ridzon, D., Moore, J.H., Characterization of microRNA expression levels and their biological correlates in human cancer cell lines (2007) Cancer Res, 67, pp. 2456-2468; Varnholt, H., Drebber, U., Schulze, F., Wedemeyer, I., Schirmacher, P., MicroRNA gene expression profile of hepatitis C virus-associated hepatocellular carcinoma (2008) Hepatology, 47, pp. 1223-1232; Jiang, J., Gusev, Y., Aderca, I., Mettler, T.A., Nagorney, D.M., Association of MicroRNA expression in hepatocellular carcinomas with hepatitis infection, cirrhosis, and patient survival (2008) Clin Cancer Res, 14, pp. 419-427; Braconi, C., Valeri, N., Gasparini, P., Huang, N., Taccioli, C., Hepatitis C virus proteins modulate microRNA expression and chemosensitivity in malignant hepatocytes (2010) Clin Cancer Res, 16, pp. 957-966; Abbas, T., Dutta, A., p21 in cancer: intricate networks and multiple activities (2009) Nat Rev Cancer, 9, pp. 400-414; Koike, K., Hepatitis C virus contributes to hepatocarcinogenesis by modulating metabolic and intracellular signaling pathways (2007) J Gastroenterol Hepatol, 22, pp. S108-S111; Darvesh, A.S., Aggarwal, B.B., Bishayee, A., Curcumin and liver cancer: a review (2012) Curr Pharm Biotechnol, 13, pp. 218-228; Srivastava, R.K., Chen, Q., Siddiqui, I., Sarva, K., Shankar, S., Linkage of curcumin-induced cell cycle arrest and apoptosis by cyclin-dependent kinase inhibitor p21(/WAF1/CIP1) (2007) Cell Cycle, 6, pp. 2953-2961; Wang, W.Z., Cheng, J., Luo, J., Zhuang, S.M., Abrogation of G2/M arrest sensitizes curcumin-resistant hepatoma cells to apoptosis (2008) FEBS Lett, 582, pp. 2689-2695; Moriya, K., Fujie, H., Shintani, Y., Yotsuyanagi, H., Tsutsumi, T., The core protein of hepatitis C virus induces hepatocellular carcinoma in transgenic mice (1998) Nat Med, 4, pp. 1065-1067; Lee, M.N., Jung, E.Y., Kwun, H.J., Jun, H.K., Yu, D.Y., Hepatitis C virus core protein represses the p21 promoter through inhibition of a TGF-beta pathway (2002) J Gen Virol, 83, pp. 2145-2151; Yamanaka, T., Uchida, M., Doi, T., Innate form of HCV core protein plays an important role in the localization and the function of HCV core protein (2002) Biochem Biophys Res Commun, 294, pp. 521-527; Nguyen, H., Mudryj, M., Guadalupe, M., Dandekar, S., Hepatitis C virus core protein expression leads to biphasic regulation of the p21 cdk inhibitor and modulation of hepatocyte cell cycle (2003) Virology, 312, pp. 245-253; Ohkawa, K., Ishida, H., Nakanishi, F., Hosui, A., Ueda, K., Hepatitis C virus core functions as a suppressor of cyclin-dependent kinase-activating kinase and impairs cell cycle progression (2004) J Biol Chem, 279, pp. 11719-11726; Chen, P.J., Lin, M.H., Tai, K.F., Liu, P.C., Lin, C.J., The Taiwanese hepatitis C virus genome: sequence determination and mapping the 5′ termini of viral genomic and antigenomic RNA (1992) Virology, 188, pp. 102-113; Beard, M.R., Abell, G., Honda, M., Carroll, A., Gartland, M., An infectious molecular clone of a Japanese genotype 1b hepatitis C virus (1999) Hepatology, 30, pp. 316-324; Livak, K.J., Schmittgen, T.D., Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method (2001) Methods, 25, pp. 402-408; Wu, S., Huang, S., Ding, J., Zhao, Y., Liang, L., Multiple microRNAs modulate p21Cip1/Waf1 expression by directly targeting its 3′ untranslated region (2010) Oncogene, 29, pp. 2302-2308; Falcón, V., Acosta-Rivero, N., Chinea, G., de la Rosa, M.C., Menéndez, I., Nuclear localization of nucleocapsid-like particles and HCV core protein in hepatocytes of a chronically HCV-infected patient (2003) Biochem Biophys Res Commun, 310, pp. 54-58; Yamaguchi, R., Momosaki, S., Gao, G., Hsia, C.C., Kojiro, M., Truncated hepatitis C virus core protein encoded in hepatocellular carcinomas (2004) Int J Mol Med, 14, pp. 1097-1100; Chen, W., Zhang, Z., Chen, J., Zhang, J., Zhang, J., HCV core protein interacts with Dicer to antagonize RNA silencing (2008) Virus Res, 133, pp. 250-258; Pedersen, I.M., Cheng, G., Wieland, S., Volinia, S., Croce, C.M., Interferon modulation of cellular microRNAs as an antiviral mechanism (2007) Nature, 449, pp. 919-922; Tang, J.T., Wang, J.L., Du, W., Hong, J., Zhao, S.L., MicroRNA 345, a methylation-sensitive microRNA is involved in cell proliferation and invasion in human colorectal cancer (2011) Carcinogenesis, 32, pp. 1207-1215; Pan, W., Zhu, S., Yuan, M., Cui, H., Wang, L., MicroRNA-21 and microRNA-148a contribute to DNA hypomethylation in lupus CD4+ T cells by directly and indirectly targeting DNA methyltransferase 1 (2010) J Immunol, 184, pp. 6773-6781; Marquez, R.T., Bandyopadhyay, S., Wendlandt, E.B., Keck, K., Hoffer, B.A., Correlation between microRNA expression levels and clinical parameters associated with chronic hepatitis C viral infection in humans (2010) Lab Invest, 90, pp. 1727-1736; Cervigne, N.K., Reis, P.P., Machado, J., Sadikovic, B., Bradley, G., Identification of a microRNA signature associated with progression of leukoplakia to oral carcinoma (2009) Hum Mol Genet, 18, pp. 4818-4829; Guled, M., Lahti, L., Lindholm, P.M., Salmenkivi, K., Bagwan, I., CDKN2A, NF2, and JUN are dysregulated among other genes by miRNAs in malignant mesothelioma -A miRNA microarray analysis (2009) Genes Chromosomes Cancer, 48, pp. 615-623; Wang, W., Peng, B., Wang, D., Ma, X., Jiang, D., Human tumor microRNA signatures derived from large-scale oligonucleotide microarray datasets (2011) Int J Cancer, 129, pp. 1624-1634

PY - 2013

Y1 - 2013

N2 - Background: Hepatitis C virus (HCV) has been reported to regulate cellular microRNAs. The HCV core protein is considered to be a potential oncoprotein in HCV-related hepatocellular carcinoma, but HCV core-modulated cellular microRNAs are unknown. The HCV core protein regulates p21Waf1/Cip1 expression. However, the mechanism of HCV core-associated p21Waf1/Cip1 regulation remains to be further clarified. Therefore, we attempted to determine whether HCV core-modulated cellular microRNAs play an important role in regulating p21Waf1/Cip1 expression in human hepatoma cells. Methods: Cellular microRNA profiling was investigated in core-overexpressing hepatoma cells using TaqMan low density array. Array data were further confirmed by TaqMan real-time qPCR for single microRNA in core-overexpressing and full-length HCV replicon-expressing cells. The target gene of microRNA was examined by reporter assay. The gene expression was determined by real-time qPCR and Western blotting. Apoptosis was examined by annexin V-FITC apoptosis assay. Cell cycle analysis was performed by propidium iodide staining. Cell proliferation was analyzed by MTT assay. Results: HCV core protein up- or down-regulated some cellular microRNAs in Huh7 cells. HCV core-induced microRNA-345 suppressed p21Waf1/Cip1 gene expression through targeting its 3′ untranslated region in human hepatoma cells. Moreover, the core protein inhibited curcumin-induced apoptosis through p21Waf1/Cip1-targeting microRNA-345 in Huh7 cells. Conclusion and Significance: HCV core protein enhances the expression of microRNA-345 which then down-regulates p21Waf1/Cip1 expression. It is the first time that HCV core protein has ever been shown to suppress p21Waf1/Cip1 gene expression through miR-345 targeting. © 2013 Shiu et al.

AB - Background: Hepatitis C virus (HCV) has been reported to regulate cellular microRNAs. The HCV core protein is considered to be a potential oncoprotein in HCV-related hepatocellular carcinoma, but HCV core-modulated cellular microRNAs are unknown. The HCV core protein regulates p21Waf1/Cip1 expression. However, the mechanism of HCV core-associated p21Waf1/Cip1 regulation remains to be further clarified. Therefore, we attempted to determine whether HCV core-modulated cellular microRNAs play an important role in regulating p21Waf1/Cip1 expression in human hepatoma cells. Methods: Cellular microRNA profiling was investigated in core-overexpressing hepatoma cells using TaqMan low density array. Array data were further confirmed by TaqMan real-time qPCR for single microRNA in core-overexpressing and full-length HCV replicon-expressing cells. The target gene of microRNA was examined by reporter assay. The gene expression was determined by real-time qPCR and Western blotting. Apoptosis was examined by annexin V-FITC apoptosis assay. Cell cycle analysis was performed by propidium iodide staining. Cell proliferation was analyzed by MTT assay. Results: HCV core protein up- or down-regulated some cellular microRNAs in Huh7 cells. HCV core-induced microRNA-345 suppressed p21Waf1/Cip1 gene expression through targeting its 3′ untranslated region in human hepatoma cells. Moreover, the core protein inhibited curcumin-induced apoptosis through p21Waf1/Cip1-targeting microRNA-345 in Huh7 cells. Conclusion and Significance: HCV core protein enhances the expression of microRNA-345 which then down-regulates p21Waf1/Cip1 expression. It is the first time that HCV core protein has ever been shown to suppress p21Waf1/Cip1 gene expression through miR-345 targeting. © 2013 Shiu et al.

KW - core protein

KW - curcumin

KW - cyclin dependent kinase inhibitor 1

KW - microRNA

KW - microRNA 345

KW - unclassified drug

KW - 3' untranslated region

KW - apoptosis

KW - article

KW - cell cycle

KW - cell proliferation

KW - cell strain

KW - cell strain Huh7

KW - controlled study

KW - down regulation

KW - gene overexpression

KW - gene targeting

KW - Hepatitis C virus

KW - hepatoma cell

KW - human

KW - human cell

KW - nonhuman

KW - p21Waf1Cip1 gene

KW - protein expression

KW - replicon

KW - upregulation

U2 - 10.1371/journal.pone.0061089

DO - 10.1371/journal.pone.0061089

M3 - Article

VL - 8

JO - PLoS One

JF - PLoS One

SN - 1932-6203

IS - 4

ER -