Our study demonstrates that staurosporine (STS), a protein kinase inhibitor from Streptomyces sp., induces apoptosis in human papillomavirus positive oral carcinoma cells (KB) in a dose dependent manner. Growth inhibition studies revealed an IC(50) value of approximately 100 nM. STS induced marked nuclear fragmentation and inter-nucleosomal cleavage compared to untreated cells. It also caused dose dependent disruption of mitochondrial membrane potential and activation of caspase-3, indicating involvement of mitochondria-mediated cell death signaling in KB cell apoptosis. We found time-dependent arrest of the KB cells at G2/M phase of cell cycle. Using fluorescence microscopy, we have further shown that STS treatment disrupts microtubules and reorganizes F-actin after 6h exposure. Taken together, our results suggest that STS induces mitochondria-mediated KB cell apoptosis at G2/M phase by altering cell cytoskeletal network.