Ribitol-5-phosphate transferase fukutin inhibits the secretion of α-dystroglycan in HeLa cells

Acta Universitatis Medicinalis Anhui 2022 10 v.57 1640-1645     font:big middle small

Found programs:

Authors:Zhou Heng; Qiao Bing; Li Caihong; Li Qiang; Zhu Fangxiu; Fan Libin

Keywords:FKTN;cell cycle;apoptosis;migration;secretion

DOI:10.19405/j.cnki.issn1000-1492.2022.10.023

〔Abstract〕 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-3 xFlag, 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 Overex-pressing FKTN arrests cell cycle and inhibits the secretion of α-DG in HeLa cells. Apoptosis and cell migration ofHeLa cells are not affected by the overexpression of FKTN.