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Inhibition Of c‐Met Expression In Regenerating Rat Liver By RNA Interference
Author(s) -
Paranjpe Shirish,
Bowen William C,
Barua Lindsay,
Michalopoulos George K,
NejakBowen Kari
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.5.a1090
Subject(s) - small hairpin rna , liver regeneration , rna interference , biology , messenger rna , rna , regeneration (biology) , gene silencing , hepatocyte growth factor , microbiology and biotechnology , kidney , hepatectomy , chemistry , receptor , gene , endocrinology , medicine , biochemistry , surgery , resection
Hepatocyte growth factor (HGF) and its receptor c‐met are important mediators of epithelial‐mesenchymal interactions that are involved in diverse processes including regeneration of several tissues such as liver, kidney and lungs. The importance of HGF‐Met signaling system during liver development has been demonstrated by knockout experiments. Targeted disruptions of both c‐met and HGF genes results in multiple developmental defects and are embryonically lethal. The role of c‐met in liver regeneration in rat following partial hepatectomy (PHx) was investigated by utilizing RNA interference to silence c‐met in regenerating rat livers. To this end, rat c‐met‐specific silencing RNA were designed to down regulate c‐met expression in regenerating rat livers. A cocktail of two c‐met specific short hairpin RNA (ShRNA) sequences, M1 and M2, were tested for their ability to down regulate c‐met expression in liver as assessed by real‐time PCR. As negative controls, a scrambled sequence and a mismatch sequence differing from c‐met sequence at two positions were used. The animals received two injections of 150ug/rat of M1 and M2 linear polyethylenimine complex via the superior mesenteric vein. The first injection was 24 hours before two‐thirds partial hepatectomy while the second injection was at the time of partial hepatectomy. Suppression of c‐met mRNA as determined by real time RT‐PCR was observed in rats treated with ShRNA as compared to scrambled and mismatch treated rats. Immunohistochemical analysis revealed a complete absence of mitosis in treated animals at 24 hours post PHx compared to animals treated with scrambled and mismatch ShRNA.