Effects of SHP-2 tyrosine phosphatase on proliferation of liver cancer cells

Objective To construct the HepG2 cell line stably transfected by SHP-2 over-expression wild-type( WT)and gain-of-function mutation( MT),and to investigate the effects of SHP-2 gain-of-Function mutation( MT) and over-expression wild-type( WT) on proliferation capacity of liver cancer cells. Methods The successfully constructed eukaryotic expression vector of SHP-2D61G / +( MT),wild-type( WT) and vector pcDNA3. 1 were transfected into HepG2 cells,meanwhile,the untransfected ones were used as controls. Western blotting was applied to detect SHP-2 protein expression levels of transfected and untransfected cells. MTT assay was used to detect the cell proliferation. Plate colony and soft-agar colony formation assay was applied to detect cell colony formation ability. Results SHP-2 mutant and wild-type eukaryotic expression vector was successfully transfected into HepG2 cells. MTT assay showed that the proliferation rate of the control group was similar to that of the no-load group and with slow proliferation,while in the mutant group and over-expression wild-type group,the cell proliferation was significantly faster. Significant differences were found between the latter two groups and the first two groups( P 0. 01). Plate colony and soft-agar colony formation experimental results show that the number of colony-forming cells in the mutation group and the over-expression wild-type group was more than that in the noload group and the control group,and with statistically significant difference( P 0. 01). Conclusions HepG2 cell line stably transfected by SHP-2 over-expression wild-type( WT) and gain-of-function mutation( MT) is successfully established. SHP-2 gain-of-function mutation and over-expression wild-type promote the proliferation capacity of liver cancer cells.