Exploring the regulation of homo-heterologous adhesion crosstalk by Qizhu Kangai Formula based on PTP1B research on the mechanism of inhibiting colon cancer metastasis.

Objective Exploring the mechanism of Qizhu Kangai Formula in regulating the key protein of protein tyrosine phosphatase 1B (PTP1B) of homo-heterologous adhesion crosstalk inhibition of colon cancer metastasis. Methods Prepare blank serum and Qizhu Kangai Formula containing serum. Analysis of effective chemical components in serum containing Qizhu Kangai Formula using liquid chromatography-mass spectrometry technology. Constructing PTP1B overexpression model(OE-PTP1B HCT116) in vitro through phasmid transfection. After detecting the cell survival rate of OE-PTP1B HCT116 using CCK-8 method, select 20% medicated serum and 5%, 10%, 20% Qizhu Kangai Formula containing serum. Scratch assay, transwell assay, and adhesion assay were used to detect the migration, invasion, and adhesion of cells overexpressing PTP1B, respectively. Western blotting was used to detect protein expression level of PTP1B, vinculin, E-cadherin, integrin αν subunit and integrin β3 subunits. Results Liquid chromatography-mass spectrometry analysis revealed that the serum of Qizhu Kangai Formula contains 12 main effective chemical components. Among them, the highest content is Calycosin-7-glucoside. The expression levels of PTP1B and vinculin proteins were increased (P<0.05), indicating the successful construction of the model. Compared with the blank serum group, the survival rate of cells overexpressing PTP1B was decreased in different volume fractions of Qizhu Kangai Formula containing serum groups (P<0.05). Compared with the blank serum group, the wound healing rate of cells in the 10% and 20% Qizhu Kangai Formula containing serum groups decreased, the number of invasive cells in each containing serum group decreased, the number of HCT116 cells adhering to cells in each Qizhu Kangai Formula containing serum group increased, the number of HCT116 cells adhering to the matrigel decreased. Compared with the blank serum group, the expression level of E-cadherin protein was increased(P<0.05), the expression level of PTP1B, vinculin, integrin αν subunit, and integrin β3 subunit were reduced in the 20% Qizhu Kangai Formula containing serum group. Conclusion The mechanism of Qizhu Kangai Formula in inhibiting colon cancer metastasis may be related to the regulation of homo- and hetero-adhesion crosstalk mediated by PTP1B. [ABSTRACT FROM AUTHOR]