title: In vitro anti-inflammatory and anti-cancer activities of Cuscuta reflexa Roxb.
abstract: OBJECTIVE
  To determine anti-inflammatory and anti-cancer activities of Cuscuta reflexa in cell lines (in vitro).
  
  
  METHODS
  Anti-inflammatory activity of the water extract was analysed in vitro using lipopolysaccharide (LPS) induced inflammatory reactions in murine macrophage cell line RAW264.7. The expression of COX-2 and TNF-α genes involved in inflammation was analysed by SQ RT-PCR. EMSA was conducted to analyse the influence of the extract on NF-κB signalling. Anti-cancer activity was analysed on Hep3B cells by MTT assay, DAPI staining, annexin V staining and SQ-RT PCR analysis of BAX, Bcl-2, p53 and survivin.
  
  
  RESULTS
  The extract down regulated LPS induced over expression of TNF-α and COX-2 in RAW264.7 cells; blocked NF-κB binding to its motifs and induced apoptosis in Hep3B cells as evidenced from MTT, DAPI staining and annexin V staining assays. The extract up regulated pro-apoptotic factors BAX and p53, and down regulated anti-apoptotic factors Bcl-2 and survivin.
  
  
  CONCLUSIONS
  The study showed that Cuscuta reflexa inhibits LPS induced inflammatory responses in RAW264.7 cells through interplay of TNF-α, COX-2 and NF-κB signalling. It induced apoptosis in Hep3B cells through the up regulation of p53, BAX and down regulation of Bcl-2 and survivin.
authors.affiliation: Bioprospecting and Molecular Pharmacology Lab, Rajiv Gandhi Centre for Biotechnology Thiruvananthapuram, Kerala 695014, India.
authors.forename: V
authors.initials: V
authors.lastname: Suresh
authors.forename: V
authors.initials: V
authors.lastname: Sruthi
authors.forename: B
authors.initials: B
authors.lastname: Padmaja
authors.forename: V V
authors.initials: VV
authors.lastname: Asha
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D002460:Cell Line; D034182:Cuscuta / Q000737:chemistry*; D024202:Electrophoretic Mobility Shift Assay; D066298:In Vitro Techniques; D051034:Inhibitor of Apoptosis Proteins / Q000378:metabolism; D051379:Mice; D016328:NF-kappa B / Q000378:metabolism; D010936:Plant Extracts / Q000494:pharmacology*; D012097:Repressor Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D000077022:Survivin; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D051028:bcl-2-Associated X Protein / Q000378:metabolism
chemical_list: D000893:Anti-Inflammatory Agents; D000972:Antineoplastic Agents, Phytogenic; C117749:Birc5 protein, mouse; D051034:Inhibitor of Apoptosis Proteins; D016328:NF-kappa B; D010936:Plant Extracts; D012097:Repressor Proteins; D000077022:Survivin; D016159:Tumor Suppressor Protein p53; D051028:bcl-2-Associated X Protein