The Role of the 3' Untranslated Region in the Post-Transcriptional Regulation of KLF6 Gene Expression in Hepatocellular Carcinoma
Abstract
:1. Introduction
2. Experimental
2.1. Construction of Plasmids
Primer | Sequence |
---|---|
KLF6 F | CGG ACG CAC ACA GGA GAA AA |
KLF6 R | CGG TGT GCT TTC GGA AGT G |
UTR1F GW | GGGG ACA AGT TTG ATC AAA AAA GCA GGC TGG GAG CAG AGA GGT GGA TCC T |
UTR1R GW | GGGG AC CAC TTT GTA CAA GAA AGC TGG GTT ACA CAG CTT ATA CAA TGG GTT ACA AAT G |
UTR2F GW | GGGG ACA AGT TTG TAC AAA AAA GCA GGC TGT GTC AAG TAG CTT GTT TTA CAC GCT AC |
UTR2R GW | GGGG AC CAC TTT GTA CAA GAA AGC TGG GTA AGG TCT ATA TGA AAG TCT CAA GGT GGC |
UTR3F GW | GGGG ACA AGT TTG TAC AAA AAA GCA GGC TGA CTG TCA GTG TTA AAA TGG AAA ACA GG |
UTR3R GW | GGGG AC CAC TTT GTA CAA GAA AGC TGG GTA TAA AGC AAA GAG CCA CAC CCA C |
UTR4F GW | GGGG ACA AGT TTG TAC AAA AAA GCA GGC TAA TTG GCA TAC CAC GGC GTG |
UTR4R GW | GGGG AC CAC TTT GTA CAA GAA AGC TGG GTT GGC AGT GAT GTC ATC TTT TAT TTT CTG |
2.2. Statistics and Software
3. Results
3.1. Bioinformatic Analysis of KLF6 3'UTR
3.2. Determination of KLF6 mRNA Half-Life
3.3. KLF6 3'UTR Is a Potent Regulator of Gene Expression
4. Discussion
5. Conclusions
Abbreviations
HCC | hepatocellular carcinoma |
KLF | Kruppel-like factor |
UTR | untranslated region |
Acknowledgments
Conflicts of Interest
References
- Black, A.R.; Black, J.D.; Azizkhan-Clifford, J. Sp1 and krüppel-like factor family of transcription factors in cell growth regulation and cancer. J. Cell. Physiol. 2001, 188, 143–160. [Google Scholar] [CrossRef]
- Benzeno, S.; Narla, G.; Allina, J.; Cheng, G.Z.; Reeves, H.L.; Banck, M.S.; Odin, J.A.; Diehl, J.A.; Germain, D.; Friedman, S.L. Cyclin-dependent kinase inhibition by the KLF6 tumor suppressor protein through interaction with cyclin D1. Cancer Res. 2004, 64, 3885–3891. [Google Scholar] [CrossRef]
- Rodríguez, E.; Aburjania, N.; Priedigkeit, N.M.; DiFeo, A.; Martignetti, J.A. Nucleo-cytoplasmic localization domains regulate Krüppel-like factor 6 (KLF6) protein stability and tumor suppressor function. PLoS One 2010, 5, e12639. [Google Scholar]
- Bieker, J.J. Krüppel-like factors: Three fingers in many pies. J. Biol. Chem. 2001, 276, 34355–34358. [Google Scholar] [CrossRef]
- Kremer-Tal, S.; Reeves, H.L.; Narla, G.; Thung, S.N.; Schwartz, M.; Difeo, A.; Katz, A.; Bruix, J.; Bioulac-Sage, P.; Martignetti, J.A.; et al. Frequent inactivation of the tumor suppressor Kruppel-like factor 6 (KLF6) in hepatocellular carcinoma. Hepatology 2004, 40, 1047–1052. [Google Scholar] [CrossRef]
- Yamashita, K.; Upadhyay, S.; Osada, M.; Hoque, M.O.; Xiao, Y.; Mori, M.; Sato, F.; Meltzer, S.J.; Sidransky, D. Pharmacologic unmasking of epigenetically silenced tumor suppressor genes in esophageal squamous cell carcinoma. Cancer Cell 2002, 2, 485–495. [Google Scholar] [CrossRef]
- Kremer-Tal, S.; Narla, G.; Chen, Y.; Hod, E.; DiFeo, A.; Yea, S.; Lee, J.-S.; Schwartz, M.; Thung, S.N.; Fiel, I.M.; et al. Downregulation of KLF6 is an early event in hepatocarcinogenesis, and stimulates proliferation while reducing differentiation. J. Hepatol. 2007, 46, 645–654. [Google Scholar] [CrossRef]
- Narla, G.; DiFeo, A.; Yao, S.; Banno, A.; Hod, E.; Reeves, H.L.; Qiao, R.F.; Camacho-Vanegas, O.; Levine, A.; Kirschenbaum, A.; et al. Targeted inhibition of the KLF6 splice variant, KLF6 SV1, suppresses prostate cancer cell growth and spread. Cancer Res. 2005, 65, 5761–5768. [Google Scholar] [CrossRef]
- Reeves, H.L.; Narla, G.; Ogunbiyi, O.; Haq, A.I.; Katz, A.; Benzeno, S.; Hod, E.; Harpaz, N.; Goldberg, S.; Tal-Kremer, S.; et al. Kruppel-like factor 6 (KLF6) is a tumor-suppressor gene frequently inactivated in colorectal cancer. Gastroenterology 2004, 126, 1090–1103. [Google Scholar] [CrossRef]
- Ito, G.; Uchiyama, M.; Kondo, M.; Mori, S.; Usami, N.; Maeda, O.; Kawabe, T.; Hasegawa, Y.; Shimokata, K.; Sekido, Y. Krüppel-like factor 6 is frequently down-regulated and induces apoptosis in non-small cell lung cancer cells. Cancer Res. 2004, 64, 3838–3843. [Google Scholar] [CrossRef]
- Cho, Y.G.; Kim, C.J.; Park, C.H.; Yang, Y.M.; Kim, S.Y.; Nam, S.W.; Lee, S.H.; Yoo, N.J.; Lee, J.Y.; Park, W.S. Genetic alterations of the KLF6 gene in gastric cancer. Oncogene 2005, 24, 4588–4590. [Google Scholar] [CrossRef]
- Camacho-Vanegas, O.; Narla, G.; Teixeira, M.S.; DiFeo, A.; Misra, A.; Singh, G.; Chan, A.M.; Friedman, S.L.; Feuerstein, B.G.; Martignetti, J.A. Functional inactivation of the KLF6 tumor suppressor gene by loss of heterozygosity and increased alternative splicing in glioblastoma. Int. J. Cancer 2007, 121, 1390–1395. [Google Scholar] [CrossRef]
- Chen, H.; Liu, X.; Lin, J.; Chen, T.; Feng, Q.; Zeng, Y. Mutation analysis of KLF6 gene in human nasopharyngeal carcinomas. Ai Zheng 2002, 21, 1047–1050. [Google Scholar]
- Bureau, C.; Péron, J.M.; Bouisson, M.; Danjoux, M.; Selves, J.; Bioulac-Sage, P.; Balabaud, C.; Torrisani, J.; Cordelier, P.; Buscail, L.; et al. Expression of the transcription factor Klf6 in cirrhosis, macronodules, and hepatocellular carcinoma. J. Gastroenterol. Hepatol. 2008, 23, 78–86. [Google Scholar] [CrossRef]
- Sirach, E.; Bureau, C.; Péron, J.M.; Pradayrol, L.; Vinel, J.P.; Buscail, L.; Cordelier, P. KLF6 transcription factor protects hepatocellular carcinoma-derived cells from apoptosis. Cell Death Differ. 2007, 14, 1202–1210. [Google Scholar] [CrossRef]
- Hanoun, N.; Bureau, C.; Diab, T.; Gayet, O.; Dusetti, N.; Selves, J.; Vinel, J.-P.; Buscail, L.; Cordelier, P.; Torrisani, J. The SV2 variant of KLF6 is down-regulated in hepatocellular carcinoma and displays anti-proliferative and pro-apoptotic functions. J. Hepatol. 2010, 53, 880–888. [Google Scholar] [CrossRef]
- Bureau, C.; Hanoun, N.; Torrisani, J.; Vinel, J.-P.; Buscail, L.; Cordelier, P. Expression and function of Kruppel like-factors (KLF) in carcinogenesis. Curr. Genomics 2009, 10, 353–360. [Google Scholar] [CrossRef]
- Song, J.; Kim, C.J.; Cho, Y.G.; Kim, S.Y.; Nam, S.W.; Lee, S.H.; Yoo, N.J.; Lee, J.Y.; Park, W.S. Genetic and epigenetic alterations of the KLF6 gene in hepatocellular carcinoma. J. Gastroenterol. Hepatol. 2006, 21, 1286–1289. [Google Scholar] [CrossRef]
- Eberhardt, W.; Doller, A.; Akool, E.-S.; Pfeilschifter, J. Modulation of mRNA stability as a novel therapeutic approach. Pharmacol. Ther. 2007, 114, 56–73. [Google Scholar] [CrossRef]
- Elkon, R.; Zlotorynski, E.; Zeller, K.I.; Agami, R. Major role for mRNA stability in shaping the kinetics of gene induction. BMC Genomics 2010, 11, e259. [Google Scholar] [CrossRef]
- Siepel, A.; Bejerano, G.; Pedersen, J.S.; Hinrichs, A.S.; Hou, M.; Rosenbloom, K.; Clawson, H.; Spieth, J.; Hillier, L.W.; Richards, S.; et al. Evolutionarily conserved elements in vertebrate, insect, worm, and yeast genomes. Genome Res. 2005, 15, 1034–1050. [Google Scholar] [CrossRef]
- Xie, X.; Lu, J.; Kulbokas, E.J.; Golub, T.R.; Mootha, V.; Lindblad-Toh, K.; Lander, E.S.; Kellis, M. Systematic discovery of regulatory motifs in human promoters and 3'UTRs by comparison of several mammals. Nature 2005, 434, 338–345. [Google Scholar] [CrossRef]
- ARE site. Available online: http://rna.tbi.univie.ac.at/cgi-bin/AREsite.cgi (accessed on 25 November 2013).
- Tsai, W.-C.; Hsu, S.-D.; Hsu, C.-S.; Lai, T.-C.; Chen, S.-J.; Shen, R.; Huang, Y.; Chen, H.-C.; Lee, C.-H.; Tsai, T.-F.; et al. MicroRNA-122 plays a critical role in liver homeostasis and hepatocarcinogenesis. J. Clin. Invest. 2012, 122, 2884–2897. [Google Scholar] [CrossRef]
- Mignone, F.; Gissi, C.; Liuni, S.; Pesole, G. Untranslated regions of mRNAs. Genome Biol. 2002, 3, 1–10. [Google Scholar]
- Vázquez-Chantada, M.; Fernández-Ramos, D.; Embade, N.; Martínez-Lopez, N.; Varela-Rey, M.; Woodhoo, A.; Luka, Z.; Wagner, C.; Anglim, P.P.; Finnell, R.H.; et al. HuR/methyl-HuR and AUF1 regulate the MAT expressed during liver proliferation, differentiation, and carcinogenesis. Gastroenterology 2010, 138, 1943–1953. [Google Scholar] [CrossRef]
- Humeau, M.; Torrisani, J.; Cordelier, P. miRNA in clinical practice: Pancreatic cancer. Clin. Biochem. 2013, 46, 933–936. [Google Scholar] [CrossRef]
- Zhang, X.; Nie, Y.; Du, Y.; Cao, J.; Shen, B.; Li, Y. MicroRNA-181a promotes gastric cancer by negatively regulating tumor suppressor KLF6. Tumor Biol. 2012, 33, 1589–1597. [Google Scholar] [CrossRef]
© 2013 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).
Share and Cite
Diab, T.; Hanoun, N.; Bureau, C.; Christol, C.; Buscail, L.; Cordelier, P.; Torrisani, J. The Role of the 3' Untranslated Region in the Post-Transcriptional Regulation of KLF6 Gene Expression in Hepatocellular Carcinoma. Cancers 2014, 6, 28-41. https://doi.org/10.3390/cancers6010028
Diab T, Hanoun N, Bureau C, Christol C, Buscail L, Cordelier P, Torrisani J. The Role of the 3' Untranslated Region in the Post-Transcriptional Regulation of KLF6 Gene Expression in Hepatocellular Carcinoma. Cancers. 2014; 6(1):28-41. https://doi.org/10.3390/cancers6010028
Chicago/Turabian StyleDiab, Thoria, Naima Hanoun, Christophe Bureau, Camille Christol, Louis Buscail, Pierre Cordelier, and Jérôme Torrisani. 2014. "The Role of the 3' Untranslated Region in the Post-Transcriptional Regulation of KLF6 Gene Expression in Hepatocellular Carcinoma" Cancers 6, no. 1: 28-41. https://doi.org/10.3390/cancers6010028
APA StyleDiab, T., Hanoun, N., Bureau, C., Christol, C., Buscail, L., Cordelier, P., & Torrisani, J. (2014). The Role of the 3' Untranslated Region in the Post-Transcriptional Regulation of KLF6 Gene Expression in Hepatocellular Carcinoma. Cancers, 6(1), 28-41. https://doi.org/10.3390/cancers6010028