Journal Article

Gold (I) N-heterocyclic carbene complex inhibits mouse melanoma growth by p53 upregulation

Das, Sujata, Saha, Krishna Das, Munda, Rudra Narayan, Dey, Sumit Kumar, Nandy, Abhishek, Dinda, Joydev

2014

Overview

Background Cancer treatment using gold (I) complexes is becoming popular. In this study, a gold (I) N-heterocyclic complex designated as complex 3 was synthesized, its cytotoxicity was examined, and its anti-melanoma activity was evaluated in vitro and in vivo . Methods Viability of cancer cells was determined by MTT assay upon treatment with various concentrations of a gold (I) N-heterocyclic carbene complex (complex 3 ) in a dose and time dependent manner. Mouse melanoma cells B16F10 were selected for further apoptotic studies, including flowcytometric analysis of annexin binding, cell cycle arrest, intracellular ROS generation and loss in the mitochondrial membrane potential. ELISA based assays were done for caspase activities and western blots for determining the expression of various survival and apoptotic proteins. Immunocytology was performed to visualize the translocation of p53 to the nucleus. B16F10 cells were inoculated into mice and post tumor formation, complex 3 was administered. Immunohistology was performed to determine the expressions of p53, p21, NF-κB (p65 and p50), MMP-9 and VEGF. Student’s t test was used for determining statistical significance. The survival rate data were analyzed by Kaplan-Meier plots. Results Complex 3 markedly inhibited the growth of HCT 116, HepG2, and A549, and induced apoptosis in B16F10 cells with nuclear condensation, DNA fragmentation, externalization of phosphatidylserine, activation of caspase 3 and caspase 9, PARP cleavage, downregulation of Bcl-2, upregulation of Bax, cytosolic cytochrome c elevation, ROS generation, and mitochondrial membrane potential loss indicating the involvement of an intrinsic mitochondrial death pathway. Further, upregulation of p53, p-p53 (ser 15) and p21 indicated the role of p53 in complex 3 mediated apoptosis. The complex reduced tumor size, and caused upregulation of p53 and p21 along with downregulation of NF-κB (p65 and p50), VEGF and MMP-9. These results suggest that it induced anti-melanoma effect in vitro and in vivo by modulating p53 and other apoptotic factors. Conclusions The gold (I) N-heterocyclic carbene complex (C 22 H 26 N 6 AuO 2 PF 6 ) designated as complex 3 induced ROS and p53 dependent apoptosis in B16F10 cells involving the mitochondrial death pathway along with suppression of melanoma tumor growth by regulating the levels of pro and anti apoptotic factors (p53, p21, NF-κB, VEGF and MMP-9).

• Publisher: BioMed Central,BioMed Central Ltd,Springer Nature B.V
• Subject: Analysis / Animals / Antineoplastic Agents - chemical synthesis / Antineoplastic Agents - pharmacology / Apoptosis / Apoptosis - drug effects / Biology / Biomedical and Life Sciences / Biomedicine / Blotting, Western / Cancer / Cancer Research / Carbon / Cell Line, Tumor / Cytotoxicity / Disease Models, Animal / Enzyme-Linked Immunosorbent Assay / Experiments / Flow Cytometry / Gold / Gold Compounds - chemical synthesis / Gold Compounds - pharmacology / Growth / HCT116 Cells / Hep G2 Cells / Heterocyclic Compounds - chemical synthesis / Heterocyclic Compounds - pharmacology / Humans / Immunohistochemistry / Ligands / Medical equipment and supplies industry / Medical test kit industry / Melanoma / Melanoma - metabolism / Methane - analogs & derivatives / Methane - chemical synthesis / Methane - pharmacology / Mice / Oncology / Physiological aspects / Prevention / Tumor Suppressor Protein p53 - drug effects / Tumor Suppressor Protein p53 - metabolism / Tumors / Up-Regulation