The Elementary Study On The Mechanism Of GnT-?a Affecting Migration And Invasion Of HCC Cells By Vimentin

Hepatocellular carcinoma is a common solid tumor in the world and has a very high mortality rate due to difficulties in the early diagnosis and limited treatment methods.Clinical studies have shown that abnormal glycosylation of glycoproteins is closely related to the migration and invasion of cancer cells.Glycosyltransferases play pivotal role in glycosylation modification.GnT-IVa is a member of the glycosyltransferase family responsible for catalyzing the formation of the ?-1,4 GlcNAc branch on the N-glycan mannose nucleus.Previous experimental studies have found that in 16 cases of HCC tissues and adjacent normal tissues,13 cases showed the high elevation in the expression of GnT-IVa than that of adjacent normal tissues.And in vitro,GnT-IVa can promote the proliferation of liver cancer cells and accelerate the progression of tumors.However,there are relatively few studies on the migration and invasion of GnT-IVa and liver cancer cells.This study found that GnT-IVa can promote the migration and invasion of liver cancer cells,so the molecular mechanism of its action was explored.In this study,qRT-PCR and Western Blot were used to detect the basal expression of GnT-IVa in the target group,and the slow-viral packaging technique was used to stabilize Sk-Hep1 and HuH7 as cells overexpressing GnT-IVa.Lectin Blot was utilized to analyze the effect of different expression levels of GnT-IVa on N-glycans in hepatoma cells.It was found that after overexpression of GnT-IVa,the number of sugar chains in the cells increased;after returning with RNAi technology,the sugar chains decreased as GnT-IVa knocked down.Then,cell scratches and Transwell chamber invasion experiments were used to detect whether different expression levels of GnT-IVa caused changes in migration and invasion levels of liver cancer cells.It was found that the stable migration and invasion of GnT-IVa in Sk-Hep1 and HuH7 cells significantly enhanced the expression of GnT-IVa,which resulted in migration and decreased invasive ability.In this study,we further explored the specific molecular mechanism of GnT-IVa onthe base of above results,and found that GnT-IVa was stably overexpressed in Sk-Hep1 and HuH7 cells,and the expression level of vimentin was significantly increased at the protein level.And the expression level of kinase Axl was significantly increased.When GnT-IVa was knocked down,the level of vimentin protein expression was significantly decreased.Then,using RNA interference technology to target knockdown of vimentin in Sk-Hep1 and HuH7 cells stably overexpressing GnT-IVa,it was observed that the expression of Axl was significantly decreased,and the migration and invasive capability of the cells was also decreased.These results suggested that GnT-IVa may affect the signaling pathway of vimentin/Axl and promote the migration and invasion of liver cancer cells.Based on the above results,this study found that GnT-IVa can enhance tumor migration and invasion ability in liver cancer cells,and GnT-IVa may regulate Axl by affecting the expression of vimentin to enhance the migration and invasion of liver cancer cells.These findings have a certain value in elucidating the important role of GnT-IVa in the process of liver cancer migration and invasion,and provide new ideas for targeting drug receptors and anti-tumor metastasis research.