Cited by 100Liver physiology and pathologyHepatocellular Carcinoma Treatment and PrognosisTissue Engineering and Regenerative Medicine

Cell cycle effects resulting from inhibition of hepatocyte growth factor and its receptor c-Met in regenerating rat livers by RNA interference

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Paranjpe, Shirish, Bowen, William C., Bell, Aaron W., Nejak-Bowen, Kari, Luo, Jian-Hua, Michalopoulos, George K.·2007·Hepatology

Abstract

Hepatocyte growth factor (HGF) and its receptor c-Met are involved in liver regeneration. The role of HGF and c-Met in liver regeneration in rat following two-thirds partial hepatectomy (PHx) was investigated using RNA interference to silence HGF and c-Met in separate experiments. A mixture of 2 c-Met-specific short hairpin RNA (ShRNA) sequences, ShM1 and ShM2, and 3 HGF-specific ShRNA, ShH1, ShH3, and ShH4, were complexed with linear polyethylenimine. Rats were injected with the ShRNA/PEI complex 24 hours before and at the time of PHx. A mismatch and a scrambled ShRNA served as negative controls. ShRNA treatment resulted in suppression of c-Met and HGF mRNA and protein compared with that in controls. The regenerative response was assessed by PCNA, mitotic index, and BrdU labeling. Treatment with the ShHGF mixture resulted in moderate suppression of hepatocyte proliferation. Immunohistochemical analysis revealed severe suppression of incorporation of BrdU and complete absence of mitosis in rats treated with ShMet 24 hours after PHx compared with that in controls. Gene array analyses indicated abnormal expression patterns in many cell-cycle- and apoptosis-related genes. The active form of caspase 3 was seen to increase in ShMet-treated rats. The TUNEL assay indicated a slight increase in apoptosis in ShMet-treated rats compared with that in controls. Conclusion: The data indicated that in vivo silencing of c-Met and HGF mRNA by RNA interference in normal rats results in suppression of mRNA and protein, which had a measurable effect on proliferation kinetics associated with liver regeneration. (Hepatology 2007.)

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  • Under shRNA conditions, at 24 h after injection, MET mRNA levels are reduced ~1.5 fold and tyrosine-phosphorylated MET protein levels are decreased ~3 fold.

    “Specifically, at 24 h after shRNA injection (when the surgeries are performed) MET mRNA levels are reduced ∼1.5 fold, and tyrosine-phosphorylated MET protein levels are decreased ∼3 fold [24].”

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Authors
Paranjpe, S., Bowen, W. C., Bell, A. W., Nejak-Bowen, K., Luo, J.-H., & Michalopoulos, G. K.
Affiliations
University of Pittsburgh (US)
Corresponding author
Michalopoulos, George K.
Venue
Hepatology
Published
2007-06
Volume
45
Issue
6
Pages
1471-1477
ISSN
0270-9139 (ISSN-L)
OpenAlex
W1984032706
Cited by
100
References
38
Citations by year
2026: 2 · 2025: 1 · 2023: 2 · 2022: 1 · 2021: 5 · 2020: 7 · 2019: 3 · 2018: 9 · 2017: 3 · 2016: 1 · 2015: 3 · 2014: 8 · 2013: 12 · 2012: 7
FWCI
7.08
Type
journal article
Language
en
Open access
Locations
  • Hepatology (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
Funders
  • National Institutes of Health (2r01ca103958-06a1, UL1TR000005)
  • National Cancer Institute (CA 35373, R01CA035373, R01-CA103958, CA103958)
  • National Center for Advancing Translational Sciences
Topics
  • Health Sciences › Medicine › Hepatology › Liver physiology and pathology
  • Health Sciences › Medicine › Hepatology › Hepatocellular Carcinoma Treatment and Prognosis
  • Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
Keywords
hepatocyte growth factor, c-Met, liver regeneration, partial hepatectomy, RNA interference, shRNA, hepatocyte proliferation, cell cycle, apoptosis, caspase-3, gene silencing
MeSH terms
Animals, Cell Cycle, Cell Division, Hepatectomy, Liver, Liver Regeneration, RNA, Messenger, Hepatocyte Growth Factor, Proto-Oncogene Proteins c-met, In Situ Nick-End Labeling, Oligonucleotide Array Sequence Analysis, RNA Interference, Green Fluorescent Proteins, Rats, Caspase 3

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