Immunoblotting analyses were performed to ascertain protein kinase C activation, EGFR status, EGFR phosphorylation and mitogen activated protein kinase (MAPK) activity. Flow cytometry analysis and ELISA reconfirmed tyrosine phosphorylation of EGFR and ERK activations, respectively. PKC-伪 siRNA knockdown was done to corroborate the involvement of PKC-伪 in the undertaken study.
Our study showed the translocation of PKC-伪 from cytosol to the membrane fraction in colon carcinoma cell lines on incubation with TDH. The EGFR tyrosine kinase activity exhibited a down regulation on TDH treatment which involved PKC-伪, as confirmed by siRNA knockdown. Also ERK phosphorylation occurred on PKC-伪 activation. Conclusion: TDH activated PKC鈥愇?down regulates EGFR tyrosine kinase activity by MEK dependent mechanism involving MAPK.
In this study we have seen that TDH has an implication in EGFR based therapeutic approach in colorectal cancer via PKC mediated mechanism.