PKI 166 induced redox signalling and apoptosis through activation of p53, MAP kinase and caspase pathway in epidermoid carcinoma.

Abstract

Cellular redox changes have emerged as a pivotal and proximal event in cancer. PKI 166 is used to determine the effects of redox sensitive inhibition of EGFR, metastasis and apoptosis in epidermoid carcinoma. Cytotoxicity study of PKI 166 (IC50 1.0 microM) treated A431 cells were performed by MTT assay for 48 and 72 hrs. Morphological analysis of PKI 166 treated A431 cells for 48 hrs. revealed the cell shrinkage, loss of filopodia and lamellipodia by phase contrast and SEM images in dose dependent manner. It has cytotoxic effects through inhibiting cellular proliferation, leads to the induction of apoptosis, as increased fraction of sub-G1 phase of the cell cycle, chromatin condensation and DNA ladder. It inhibited cyclin-D1 and cyclin-E expression and induced p53, p21 expression in dose dependent manner. Consequently, an imbalance of Bax/Bcl-2 ratio triggered caspase cascade and subsequent cleavage of PARP, thereby shifting the balance in favour of apoptosis. PKI 166 treatment actively stimulated reactive oxygen species (ROS) and mitochondrial membrane depolarization. It inhibited some metastatic properties of A431 cells supressing colony formation by soft agar assay and inhibition of MMP 9 activity by gelatin zymography and western blot analysis. PKI 166 inhibited growth factor induced phosphorylation of EGFR, Akt, MAPK, JNK and colony formation in A431 cells. Thus the inhibition of proliferation was associated with redox regulation of the caspase cascade, EGFR, Akt/PI3K, MAPK/ ERK and JNK pathway. On the other hand, increased antioxidant activity leads to decreased ROS generation inhibit the anti-proliferative and apoptotic properties of PKI 166 in A431 cells. These observations indicated PKI 166 induced redox signalling dependent inhibition of cell proliferation, metastatic properties and induction of apoptotic potential in epidermoid carcinoma.

• Authors: Subhasis Das, Kaushik Kumar Dey, Rashmi Bharti, Sujata MaitiChoudhury, Sukumar Maiti, Mahitosh Mandal
• Journal: Journal of experimental therapeutics & oncology (J Exp Ther Oncol) Vol. 10 Issue 2 Pg. 139-53 ( 2012) ISSN: 1359-4117 [Print] United States
• PMID: 23350354 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Pyrimidines
• Pyrroles
• Reactive Oxygen Species
• TP53 protein, human
• Tumor Suppressor Protein p53
• Cytochromes c
• PKI 166
• Catalase
• Superoxide Dismutase
• Phosphatidylinositol 3-Kinases
• EGFR protein, human
• ErbB Receptors
• Proto-Oncogene Proteins c-akt
• Mitogen-Activated Protein Kinases
• Caspases

Topics

• Apoptosis (drug effects)
• Blotting, Western
• Carcinoma, Squamous Cell (drug therapy, metabolism, pathology)
• Caspases (metabolism)
• Catalase (metabolism)
• Cell Cycle (drug effects)
• Cell Proliferation (drug effects)
• Cytochromes c (metabolism)
• ErbB Receptors (antagonists & inhibitors, metabolism)
• Humans
• MAP Kinase Signaling System
• Membrane Potential, Mitochondrial (drug effects)
• Mitochondria (drug effects, metabolism)
• Mitogen-Activated Protein Kinases (metabolism)
• Oxidation-Reduction
• Phosphatidylinositol 3-Kinases (metabolism)
• Proto-Oncogene Proteins c-akt (metabolism)
• Pyrimidines (pharmacology)
• Pyrroles (pharmacology)
• Reactive Oxygen Species (metabolism)
• Signal Transduction (drug effects)
• Superoxide Dismutase (metabolism)
• Tumor Cells, Cultured
• Tumor Suppressor Protein p53 (metabolism)