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RNA stability regulates differential expression of the metastasis protein, osteopontin, in hepatocellular cancer.

Publication ,  Journal Article
Emani, S; Zhang, J; Guo, L; Guo, H; Kuo, PC
Published in: Surgery
June 2008

BACKGROUND: Osteopontin (OPN) is a potential therapeutic target in hepatocellular carcinoma (HCC), because it is a critical mediator of metastatic function. The molecular mechanisms that determine expression of OPN in HCC, however, are unknown. In this study, we examine differential OPN expression in the 2 HCC cell lines: HepG2 and Hep3B. METHODS: OPN expression, metastatic function, OPN promoter activity, and active transcription of OPN mRNA and its decay were assessed in the 2 HCC cell lines using standard techniques. RNA gel-shift assays were performed to determine binding of cytoplasmic proteins to OPN mRNA. RESULTS: Expression of OPN cellular/secreted protein and mRNA was greater in HepG2 than Hep3B cells (P < .01). Transient transfection of the OPN promoter construct demonstrated equivalent luciferase activities in the 2 cell lines; the rate of transcription was also equivalent as determined by chromatin immuno-precipitation assay. OPN mRNA half-life was 21 +/- 1 h and 3 +/- 1 h in HepG2 and Hep3B, respectively (P < .02). In HepG2 and Hep3B, the nucleotide sequence of OPN and its 5'-UTR, 3'-UTR, and poly (A) tail lengths were identical. A luciferase construct coupled in line with OPN-5'-UTR and OPN 3'-UTR presented greater expression in HepG2 (P < .01 vs Hep3B). Deletion of nt 10-57 of the OPN 5'-UTR restored luciferase and HA-tagged OPN protein expression in Hep3B but not in Hep G2. RNA gel-shift assays demonstrate different patterns of protein binding to OPN 5'-UTR between the 2 HCC cell lines. CONCLUSIONS: We conclude that RNA stability is a new, previously unrecognized mechanism that regulates OPN expression in HCC to convey metastatic function.

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Published In

Surgery

DOI

EISSN

1532-7361

Publication Date

June 2008

Volume

143

Issue

6

Start / End Page

803 / 812

Location

United States

Related Subject Headings

  • Surgery
  • RNA, Messenger
  • RNA Stability
  • Protein Binding
  • Promoter Regions, Genetic
  • Phenotype
  • Osteopontin
  • Liver Neoplasms
  • Humans
  • Half-Life
 

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Emani, S., Zhang, J., Guo, L., Guo, H., & Kuo, P. C. (2008). RNA stability regulates differential expression of the metastasis protein, osteopontin, in hepatocellular cancer. Surgery, 143(6), 803–812. https://doi.org/10.1016/j.surg.2008.02.005
Emani, Sirisha, Jinping Zhang, Lucie Guo, Hongtao Guo, and Paul C. Kuo. “RNA stability regulates differential expression of the metastasis protein, osteopontin, in hepatocellular cancer.Surgery 143, no. 6 (June 2008): 803–12. https://doi.org/10.1016/j.surg.2008.02.005.
Emani, Sirisha, et al. “RNA stability regulates differential expression of the metastasis protein, osteopontin, in hepatocellular cancer.Surgery, vol. 143, no. 6, June 2008, pp. 803–12. Pubmed, doi:10.1016/j.surg.2008.02.005.
Emani S, Zhang J, Guo L, Guo H, Kuo PC. RNA stability regulates differential expression of the metastasis protein, osteopontin, in hepatocellular cancer. Surgery. 2008 Jun;143(6):803–812.
Journal cover image

Published In

Surgery

DOI

EISSN

1532-7361

Publication Date

June 2008

Volume

143

Issue

6

Start / End Page

803 / 812

Location

United States

Related Subject Headings

  • Surgery
  • RNA, Messenger
  • RNA Stability
  • Protein Binding
  • Promoter Regions, Genetic
  • Phenotype
  • Osteopontin
  • Liver Neoplasms
  • Humans
  • Half-Life