Modulation of adenylate cyclase signaling in association with MKK3/6 stabilization under combination of SAC and berberine to reduce HepG2 cell survivability

Apoptosis. 2017 Nov;22(11):1362-1379. doi: 10.1007/s10495-017-1407-x.

Abstract

Cancer cells often have faulty apoptotic pathways resulting in sustenance of survivability, tumour metastasis and resistance to anticancer drugs. Alternate strategies are sought to improve therapeutic efficacy and therefore HepG2 cells were treated with S-allyl-cysteine (SAC) and berberine (BER) to analyze their mechanistic impact upon necroptosis along with its interacting relationship to apoptosis. In the present study we observed that SAC and BER exposure reduced NFκβ nuclear translocation through adenylate cyclase-cAMP-protein kinaseA axis and eventually evaded c-FLIP inhibition. Effective RIP1 k63-polyubiquitination and persistent MKK3/MKK6 expression during drug treatment potentiated caspase8 activity via p53-DISC conformation. Resultant tBid associated lysosomal protease mediated AIF truncation induced DNA fragmentation and persuaded effector caspase mediated scramblase activation resulting induction of necroptosis in parallel to apoptotic events. SAC+BER effectively reduced Rb-phosphorylation resulting insignificant nuclear E2F presence led to ending of cell proliferation. Therefore necroptosis augmented the drug response and may be targeted alongside cell proliferation inhibition in formation of efficient therapeutics against liver cancer.

Keywords: Adenylate cyclase; CathepsinB; RIP1-K63-polyubiquitination; Rb phosphorylation; tAIF.

MeSH terms

  • Adenylyl Cyclases / genetics*
  • Adenylyl Cyclases / metabolism
  • Apoptosis / drug effects
  • Apoptosis Inducing Factor / genetics
  • Apoptosis Inducing Factor / metabolism
  • BH3 Interacting Domain Death Agonist Protein / genetics
  • BH3 Interacting Domain Death Agonist Protein / metabolism
  • Berberine / pharmacology*
  • CASP8 and FADD-Like Apoptosis Regulating Protein / genetics
  • CASP8 and FADD-Like Apoptosis Regulating Protein / metabolism
  • Caspase 8 / genetics
  • Caspase 8 / metabolism
  • Cell Line
  • Cyclic AMP / metabolism
  • Cyclic AMP-Dependent Protein Kinases / genetics
  • Cyclic AMP-Dependent Protein Kinases / metabolism
  • Cysteine / analogs & derivatives*
  • Cysteine / pharmacology
  • DNA Fragmentation / drug effects
  • Drug Combinations
  • Gene Expression Regulation
  • Hep G2 Cells
  • Humans
  • Leukocytes, Mononuclear / cytology
  • Leukocytes, Mononuclear / drug effects
  • Leukocytes, Mononuclear / metabolism
  • MAP Kinase Kinase 3 / genetics*
  • MAP Kinase Kinase 3 / metabolism
  • MAP Kinase Kinase 6 / genetics*
  • MAP Kinase Kinase 6 / metabolism
  • NF-kappa B / genetics
  • NF-kappa B / metabolism
  • Necrosis / chemically induced
  • Necrosis / genetics
  • Necrosis / metabolism
  • Nuclear Pore Complex Proteins / genetics
  • Nuclear Pore Complex Proteins / metabolism
  • Protein Transport
  • RNA-Binding Proteins / genetics
  • RNA-Binding Proteins / metabolism
  • Signal Transduction*
  • Tumor Suppressor Protein p53 / genetics
  • Tumor Suppressor Protein p53 / metabolism

Substances

  • AGFG1 protein, human
  • AIFM1 protein, human
  • Apoptosis Inducing Factor
  • BH3 Interacting Domain Death Agonist Protein
  • BID protein, human
  • CASP8 and FADD-Like Apoptosis Regulating Protein
  • CFLAR protein, human
  • Drug Combinations
  • NF-kappa B
  • Nuclear Pore Complex Proteins
  • RNA-Binding Proteins
  • Tumor Suppressor Protein p53
  • Berberine
  • S-allylcysteine
  • Cyclic AMP
  • Cyclic AMP-Dependent Protein Kinases
  • MAP Kinase Kinase 3
  • MAP Kinase Kinase 6
  • MAP2K3 protein, human
  • MAP2K6 protein, human
  • CASP8 protein, human
  • Caspase 8
  • Adenylyl Cyclases
  • Cysteine