title: A novel third generation bisphosphonate, minodronate (YM529), prevented proliferation and migration of hepatocellular carcinoma cells through inhibition of mevalonate pathway.
abstract: OBJECTIVE
  Skeletal metastases and bone metasitasis are a common occurrence in patients with advanced hepatocellular carcinoma (HCC). Bisphosphonates (BPs), which are used for the treatment of osteoporosis and tumor-associated hypercalcemia, have recently been reported to decrease skeletal morbidity in patients with metastatic bone disease. Several studies revealed that nitrogen-containing BPs (N-BPs) could inhibit tumor growth and migration, indicating the possibility that N-BPs have direct inhibitory effects. We aimed to determine the effects of novel a N-BP (YM529) on human HCC cells in vitro.
  
  
  METHODS
  HCC cells were treated with various concentrations of YM529 and the growth inhibition rate was determined. Apoptosis was evaluated by caspase-3/7 assay and caspase-9 cleavage detection. The effects of YM529 on the migration of HCC cells induced by hepatocyte growth factor (HGF) were determined by cell migration assay. To evaluate the involvement of the mevalonate pathway, farnesol (FOH) and geranylgeraniol (GGOH) were added.
  
  
  RESULTS
  YM529 inhibited the proliferation of HCC cells in a dose-dependent manner. The activation of caspase-3/7 and cleavage of caspase-9 demonstrated the involvement of apoptosis in cytotoxicity. GGOH reduced the growth inhibitory effect of YM529 and suppressed the induction of caspase-3/7 activities by YM529 on HCC cells. YM529 inhibited tumor cell migration induced by HGF and this effect was reduced by co-treatment with GGOH.
  
  
  CONCLUSIONS
  YM529 inhibited the cell proliferation and migration of HCC cells, implicating the involvement of the mevalonate pathway. These results suggest that N-BPs are potential agents for the treatment of HCC skeletal metastases.
authors.affiliation: Division of Gastroenterology, Tohoku University Graduate School of Medicine, Aobaku, Sendai, Japan.
authors.forename: Takayuki
authors.initials: T
authors.lastname: Kogure
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Ueno
authors.forename: Osamu
authors.initials: O
authors.lastname: Kimura
authors.forename: Yasuteru
authors.initials: Y
authors.lastname: Kondo
authors.forename: Jun
authors.initials: J
authors.lastname: Inoue
authors.forename: Koji
authors.initials: K
authors.lastname: Fukushima
authors.forename: Takao
authors.initials: T
authors.lastname: Iwasaki
authors.forename: Tooru
authors.initials: T
authors.lastname: Shimosegawa