Expression of human reconstructed caspase 6 gene in Hep-2 cells

AIM: To observe the effect of human reconstructed caspase 6 ( rcasp 6) gene on the transfected Hep 2 cell line and its expression in the Hep 2 cell line. METHODS: Human caspase 6 full length cDNA was obtained through RT PCR, and was reconstruted into reversed version of caspase 6 by settling the small subunit prior to the large one through PCR. The gene was cloned into vector pCMV and transfected into Hep 2 cells. Expression of rcasp6 protein was detected by immunofluorescent and immunocytochemical staining. Morphologic changes of Hep 2 cells subjected to transfection of rcasp6 gene were detected by HE staining and electron microscopy. RESULTS: Human reconstructed caspase 6 gene ( rcasp6 ) was cloned and inserted into pCMV vector. The Rcasp6 protein was detected in the cytoplasm of transfected Hep 2 cells. Many cells shrinked, and some cells were killed. Under electron microscope, those shrinked cells displayed typical features of apoptosis. CONCLUSION: Expression of human Rcasp6 protein in Hep 2 cells leads to apoptosis.