title: A synthetic dsRNA, as a TLR3 pathwaysynergist, combined with sorafenib suppresses HCC in vitro and in vivo.
abstract: BACKGROUND
  Recent studies have demonstrated that synthetic dsRNAs may produce therapeutic effects in a target-independent manner through stimulation of the toll-like receptor-3 (TLR3)/interferon pathway; as a result, angiogenesis and proliferation of tumor cells are inhibited. Thus, this pathway may become a potential target of dsRNA in tumor suppression. In this study, we evaluated the role of synthetic dsRNA as a TLR3 synergist and by combining with sorafenib in anti-hepatocellular carcinoma (HCC) in vitro and in vivo.
  
  
  METHODS
  Four dsRNAs were designed and synthesized. One of them that was capable of activating TLR3 most effectively in human HCC cell line (HepG2.2.15) was selected as a TLR3 synergist (called BM-06). Subsequently, the expression of proteins relating to TLR3 signaling pathway, such as NF-κB, caspase 8 survivin, bcl-2 and PCNA affected by BM-06, sorafenib alone or in combination, was compared. The migration, proliferation and apoptosis of HepG2.2.15 cells were evaluated in presence of BM-06, sorafenib alone or in combination of both. The similar treatments were also applied in an SD rat primary HCC model.
  
  
  RESULTS
  qRT-PCR data showed that the expression of TLR3 and NF-κB in HepG2.2.15 cells was enhanced. BM-06 was selected as a TLR3 synergist capable of activating the TLR3/interferon pathway most effective among 4 synthetic dsRNAs. The migration and proliferation were significantly inhibited in treated HepG2.2.15 cells with BM-06 or Sorafenib alone as compared with PBS-sham control (P<0.01). However, the role of combination BM-06 with Sorafenib was the most prominent. Tumor cell apoptotic rate was increased by BM-06 or combination when compared to PBS or poly(I:C) (P<0.05). Similarly, in orthotopic HCC SD rats, the effect of the combination was superior to either agent alone on the inhibition of tumor growth and induction of HCC cell apoptosis (P<0.05).
  
  
  CONCLUSIONS
  dsRNA alone was capable of inhibiting the proliferation of HepG2.2.15 cells and tumor growth of orthotopic HCC SD rats, but the effect of combination of dsRNA with sorafenib was more prominent. These findings implicate the potential role of combined use of a dsRNA, a TLR3 synergist, and sorafenib in inhibition of HCC.
authors.forename: Yu-Yin
authors.initials: YY
authors.lastname: Xu
authors.affiliation: Department of Pathological Anatomy, Nantong University, Nantong, China. bl1@ntu.edu.cn.
authors.forename: Li
authors.initials: L
authors.lastname: Chen
authors.forename: Gui-Lan
authors.initials: GL
authors.lastname: Wang
authors.forename: Jia-Ming
authors.initials: JM
authors.lastname: Zhou
authors.forename: Yi-Xin
authors.initials: YX
authors.lastname: Zhang
authors.forename: Yin-Ze
authors.initials: YZ
authors.lastname: Wei
authors.forename: Yuan-Yuan
authors.initials: YY
authors.lastname: Zhu
authors.forename: Jing
authors.initials: J
authors.lastname: Qin
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000008:administration & dosage / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy / Q000235:genetics* / Q000473:pathology*; D053181:Caspase 8 / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D004195:Disease Models, Animal; D015870:Gene Expression; D006801:Humans; D007371:Interferon-gamma / Q000235:genetics / Q000378:metabolism; D008113:Liver Neoplasms / Q000188:drug therapy / Q000235:genetics* / Q000473:pathology*; D008297:Male; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism; D009536:Niacinamide / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000494:pharmacology; D010671:Phenylurea Compounds / Q000008:administration & dosage / Q000494:pharmacology*; D012330:RNA, Double-Stranded / Q000008:administration & dosage / Q000235:genetics* / Q000378:metabolism; D051381:Rats; D000077157:Sorafenib; D051196:Toll-Like Receptor 3 / Q000235:genetics* / Q000378:metabolism; D015533:Transcriptional Activation; D047368:Tumor Burden / Q000187:drug effects / Q000235:genetics
chemical_list: D000970:Antineoplastic Agents; C574691:BM-06; D016328:NF-kappa B; D010671:Phenylurea Compounds; D012330:RNA, Double-Stranded; D051196:Toll-Like Receptor 3; D009536:Niacinamide; D007371:Interferon-gamma; D000077157:Sorafenib; D053181:Caspase 8