Ras-association domain family protein 6 induces apoptosis via both caspase-dependent and caspase-independent pathways

The RASSF6 Tumor Suppressor Protein Regulates Apoptosis and the Cell Cycle via MDM2 Protein and p53 Protein

“…Apoptosis Detection Assays-Cell survival and cell death were analyzed as described previously (8,13,16). In brief, control GFP or GFP-RASSF6 was expressed in U2OS, HCT116 p53 ϩ/ϩ , and HCT116 p53 Ϫ/Ϫ cells.…”

“…However, our understanding, at the mechanistic level, of the tumor suppressive role of RASSF6 is limited. When RASSF6 is exogenously expressed, it induces apoptosis in various cells in caspase-dependent and caspase-independent manners (5,8,9). RASSF6 is implicated in tumor necrosis factor ␣-induced apoptosis in HeLa cells, in okadaic acid-induced apoptosis in rat hepatocytes, and in sorbitol-induced apoptosis in human renal proximal tubular epithelial cells.…”

RASSF6 (Ras association (RalGDS/AF-6) domain family member 6)

“…There are some inconsistencies in the literature regarding which may be the major isoform, however all functional work to date has studied the larger 369 amino acid protein, which is hereafter referred to as RASSF6. Using an in-house antibody Ikeda et al, (2007) were unable to detect endogenous RASSF6 protein. All functional work has been performed using overexpressed RASSF6 protein.…”

“…There has been no systematic analysis of the expression patterns of the different RASSF6 isoforms. RT-PCR analysis of a range of tumour cell lines showed that RASSF6 is highly expressed in HeLa and A549 cells with lower levels observed in MCF-7, U373, H1299 and HepG2 (Ikeda et al, 2007). It appears from these studies that the tissue distribution of RASSF6 expression is much more restricted than that of the RASSF members RASSF1, RASSF2 and RASSF5.…”

Threonine 74 of MOB1 is a putative key phosphorylation site by MST2 to form the scaffold to activate nuclear Dbf2-related kinase 1

“…cDNAs of human MOB1 and NDR1 were obtained from human lung cDNA library (BioChain Institute Inc., Hayward, CA, USA) by PCR using the primer sets (5 0 -acgcgtatgagcttcctcttcagcag-3 0 and 5 0 -gtcgactatctgtcttttgatc caag-3 0 for MOB1; and 5 0 -acgcgtgccatggcaatgacaggctcaac-3 0 and 5 0 -gtcgactattttgctgctttcatgta-3 0 for NDR1). pCIneoMyc, pCIneoFH and pGex4T-1 (GE Healthcare Bio-sciences, UK) vectors were used to generate expression constructs (Ikeda et al, 2007). MST2, MOB1 and NDR1 expression constructs cover the following regions; pCIneoMyc and pGex4T-1 MOB1 2-216 (AAH03398); pCIneoMyc, pCIneoFH and pGex4T-1 MST2, 2-491 (AAH10640); pCIneoMyc and pCIneoFLAG NDR1, 1-465 (AAH12085).…”