排闼消癥散通过调控MEK/ERK信号通路对结肠癌侵袭转移的影响OA

Objective It is to explore the effect and mechanism of Paita Xiaozheng Powder(PXP)in inhibiting the in-vasion and metastasis of colon cancer.Methods ①Twenty male SD rats were randomly divided into 4 groups,with 5 rats in each group.The low-dose,medium-dose and high-dose groups of PXP were administered 2 mL of PXP concentrated solu-tion at doses of 0.5 g/mL,1 g/mL,and2 g/mL via gavage,respectively,the control group was administered an equal vol-ume of distilled water via gavage,the serum samples were collected after 7 days of continuous gavage.HCT-116 cells(a human colon cancer cell line)in the logarithmic growth phase were seeded in 24-well plates and then divided into 5 groups:the control group was treated with 10%serum of rats in the control group for 24 h,the low-dose,medium-dose and high-dose groups of PXP were respectively treated with 10%serum containing the drug of rats from the low-dose,medium-dose and high-dose groups of PXP for 24 h,the high-dose PXP+ML099 group was treated with 10%serum of rats from the high-dose group of PXP and 20 μmol/L ML099,a mitogen-activated protein kinase(MEK)/extracellular signal-regulated kinase(ERK)signaling pathway activator,for 24 h.The proliferative activity of HCT-116 cells was detected by CCK-8 as-say;the migration and invasion capabilities of HCT-116 cells were evaluated by cell scratch assay and the Transwell assay,and the protein expressions of metastasis-related proteins in HCT-116 cells,including E-cadherin,N-cadherin,vimentin,and MEK/ERK signaling pathway proteins were detected by Western blot.②Twelve BALB/c nude mice were prepared with HCT-116 cell implantation method to establish colon cancer xenograft models.Once the tumors had developed,the nude mice were randomly divided into 4 groups,with 3 mice in each group.The low-dose,medium-dose and high-dose groups of PXP were administered 2.53 g/kg,5.06 g/kg,and 10.12 g/kg of PXP via gavage,respectively,the control group was given an equal volume of distilled water via gavage,all once daily,continuously treated until 35 days after HCT-116 cell implantation.During the gavage period,the volume of the colon cancer xenograft was measured once every 7 days.In 35 days after implantation,the colon cancer xenograft tissue was harvested and weighed,and the expressions of proteins in the MEK/ERK signaling pathway in the xenograft tissue were detected by Western blot and immunohistochemistry.Results ①Compared with the control group,the cell proliferation activities and scratch healing rates of HCT-116 cells in each dose groups of PXP,as well as the number of migrating and invading cells were significantly decreased(all P<0.05),the rela-tive protein expressions of E-cadherin in the cells were significantly increased(all P<0.05),while the relative protein ex-pressions of N-cadherin and vimentin,as well as the p-MEK/MEK and p-ERK/ERK ratios were significantly decreased(all P<0.05).Compared with the high-dose PXP group,the cell proliferation activity and scratch healing rate of HCT-116 cells in the high-dose PXP+ML099 group,as well as the number of migrating and invading cells were significantly in-creased(all P<0.05),the relative protein expression of E-cadherin in the cells was significantly reduced(P<0.05),while the relative protein expressions of N-cadherin and vimentin,as well as the p-MEK/MEK and p-ERK/ERK ratios were significantly increased(all P<0.05).②Compared with the control group,the volume and tumor weight of colon cancer xenografts in each dose group of PXP were significantly reduced(all P<0.05),and the p-MEK/MEK and p-ERK/ERK ratios,as well as the percentages of p-MEK-and p-ERK-positive cells in the colon cancer xenograft tissue were all signifi-cantly reduced(all P<0.05).Conclusion PXP can inhibit the invasion and metastasis of colon cancer,and its mecha-nism of action may be related to the regulation of MEK/ERK signaling pathway.