Experimental Study On The Pine Pollen Aqueous Extract Inhibiting Hepatocellular Carcinoma Cells By Regulating The Expression Of ENO1

Objective: To investigate how pine pollen(PP)aqueous extract can inhibit the over activation of phosphatidylinositol-3-kinase / protein serine threonine kinase(PI3K / AKT)pathway by affecting the expression product of enolase 1(ENO1)gene,thereby inhibiting the proliferation,invasion and migration of liver cancer cells.Methods: ⑴ In this study,we first analyzed the bioinformatics of ENO1.⑵Detect the main anticancer components in the water extract of PP.⑶ Cell counting kit 8(CCK8)assay,Transwell assay and Western blot assay were performed on SMMC-7721 cells and ENO1 overexpressing stable line,respectively.Cell scratch test was performed on ENO1 overexpressing stable line.⑷ ENO1 inhibitor was used to inhibit the proliferation of ENO1 overexpressing stable line.⑸ The tumor models of nude mice with overexpression of ENO1 stably transfected strain and empty transfected cells were established.The serum of the surviving nude mice in each group was detected for alanine aminotransferase(ALT),aspartate aminotransferase(AST)and AST / ALT.The tumor blocks of the surviving nude mice were weighed and extracted for Western blot.Results: ⑴ The phosphorylation level of ENO1 in hepatocellular carcinoma(HCC)was significantly different from that in normal tissues.⑵ The contents of the main anticancer components such as total polysaccharides and flavonoids in PP and the concentration of zinc ion in aqueous extract of PP were higher than those of other antitumor drugs.⑶ PP aqueous extract inhibited the invasion and proliferation related protein expression of SMMC-7721 cells,stable transfected cells overexpressing ENO1,and empty transfected cells in vitro.PP aqueous extract inhibited the migration of stably transfected cells overexpressing ENO1 more than empty transfected cells.⑷ENO1 inhibitors significantly inhibited the survival rate of stable transfected cells overexpressing ENO1.⑸ PP aqueous extract inhibited the increase of serum ALT,AST,AST / ALT and reduced the weight of tumor in nude mice;PP aqueous extract inhibited the expression of cell proliferation-related proteins in tumor block of overexpressing ENO1 in vivo;The expression of proliferation related proteins in over expressed ENO1 tumor block was higher than that in empty transfected tumor block even under the same concentration of PP aqueous extract.Conclusion: ⑴ PP aqueous extract can inhibit the proliferation,invasion and migration of SMMC-7721 cells.⑵ The inhibition of PP aqueous extract on hepatoma cells is related to the inhibition of ENO1 and PI3 K / Akt pathway.⑶ PP aqueous extract inhibits HCC by regulating the expression of ENO1 and MBP-1 and inhibiting PI3 K / Akt pathway by inhibiting C-MYC and Erb-B2 Receptor Tyrosine Kinase 2(ERBB2),respectively.