Objective : To investigate the effects of fukutin ( FKTN) which is a member of ribitol⁃5⁃phosphate transferases on HeLa cell cycle, apoptosis, migration and the secretion of α⁃dystroglycan(α⁃DG) . Methods : The plasmid pcDNA3. 1⁃α⁃DAG⁃HA was constructed. HeLa cells were transfected with plasmid pcDNA3. 1⁃FKTN ⁃ 3xFlag, then the total protein was extracted for Western blot to determine the expression level of FKTN. Cell cycle and apoptosis were measured by flow cytometry after the overexpression of FKTN. Following the overexpression of FKTN, wound⁃healing assay was performed to detect the cell migration rate, as the same time wheat germ agglutinin⁃agarose (WGA⁃agarose) was used to enrich α⁃DG in the cell cultures, then α⁃DG was detected by Western blot. Results : The eukaryotic expression plasmid pcDNA3. 1⁃α⁃DAG⁃HA was constructed successfully. FKTN could be overexpressed in HeLa cells. After the overexpression of FKTN,the percentage of S phase of cell cycle in the experimental group decreased (P < 0. 001) and apoptosis rate unchanged (P > 0. 05) when compared with the control group. There was no change in the cell migration rate of experimental group (P > 0. 05), but after the overexpression of FKTN, secretion of α⁃DG decreased when compared with control group. Conclusion Overexpressing FKTN arrests cell cycle and inhibits the secretion of α⁃DG in HeLa cells. Apoptosis and cell migration of HeLa cells are not affected by the overexpression of FKTN.

Objective : To investigate the interaction between intracellular chloride ion protein 1(CLIC1) and activated protein kinase C receptor 1 ( RACK1) . Methods : Plasmids pcDNA3. 1⁃RACK1⁃HA and/or pcDNA3. 1 ⁃CLIC1⁃FLAG were transfected into HEK 293T cells, and pcDNA3. 1⁃RACK1⁃HA and/or pcDNA3. 1⁃CLIC1⁃FLAG were transfected into COS7. GST⁃pulldown and immunoprecipitation assays were performed to determine the interaction between CLIC1 and RACK1 in vivo and in vitro. The co⁃localization of CLIC1 and RACK1 was observed by indirect immunofluorescence assay. Results : Western blot confirmed that CLIC1 and RACK1 could be highly expressed in HEK 293T cells. GST⁃pulldown showed that RACK1 bound CLIC1 in vitro, and immunoprecipitation showed that CLIC1 and RACK1 interacted in vivo. Furthermore, indirect immunofluorescence assay showed CLIC1 co⁃localized with RACK1 . Conclusion Human CLIC1 protein interacts with RACK1 in vitro and in vivo.