Effects Of The Modified Tongyou Decoction On Vasculogenic Mimicry Of Esophageal Cancer Eca109 Cells Induced By Anoxia Via MTOR/HIF-1? Signal Pathway

Objectives To study the inhibitory effect of Modified Tongyou Decoction(MTD)on angiogenesis mimicry through m TOR/HIF-1? pathway in Eca109 cells of esophageal carcinoma induced by hypoxia.Methods Animal Experiment,By intragastric administration in rats with MDT,They were randomly divided into two groups: the control group and the drug administration group.Sera were isolated as intervention factor for cells.Cell Experiment,Hypoxia model was established with Eca109 cells.The experimental cells group were as follow: the controlled group,the anoxic group,the serum-controlled group and the serum-treatment group.Cell morphology was observed under three-dimensional cell culture(3DCC).Inhibitory effect of MTD on cell growth by Cell Counting Kit-8(CCK-8)method.Correlation between m TOR and HIF-1?,VE-cadherin and HIF-1? was detected through immunofluorescence technique.Changes of protein expression in m TOR,HIF-1? and vasculogenic mimicry molecule VE-cadherin,MMP-2 and MMP-9 in each group of Eca109 cells were analyzed by Western Blot.Expression of m RNA of m TOR,HIF-1?,VE-cadherin,MMP-2 and MMP-9 in each group was assayed by RT-PCR.Results 1 3DCC observation,the control group was lumpy,anoxic group was tubular,the anoxic group was dispersed and had apoptosis,the serum-treatment group showed mass shape and apoptosis.2 The CCK-8 assay showed that the proliferation of cells in each group was significantly different.Compared with control group,the anoxic group and the serum-controlled group,the cell proliferation was decreased compared with control group,the anoxic group and the serum-controlled group.The difference was statistically significant(P =0.000).3 Immunofluorescence results showed that m TOR/ HIF-1? and VE-cadherin / HIF-1? co-expressed in cells.4 Western blot detection that the expression of m TOR,HIF-1 alpha,VE-cadherin,MMP-2 and MMP-9 increased in the anoxic group compared with the control group(P<0.05),there was no significant difference between control group and the serum-controlled group(P > 0.05)in VE-cadherin and mmp-2(P > 0.05)and increased expression of HIF-1 alpha and MMP-9(P < 0.05).The expressions of m TOR,HIF-1?,VE-cadherin,MMP-2 and MMP-9 were downregulated in the serumtreatment group compared with the anoxic group.5 RT-PCR detection: the m TOR,HIF-1,VE-cadherin,MMP-2 and MMP-9 gene expression activities in the serum-treatment group were significantly lower than those in the anoxic group.Conclusions 1 Hypoxia can induce the formation of Eca 109 cells VM,which can increase the molecular expression of m TOR/HIF-1?.2 Modified Tongyou Decoction can significantly inhibit the formation of Eca 109 cell VM in hypoxia microenvironment.3 Under the condition of hypoxia,Modified Tongyou Decoction can significantly reduce the molecular expression of Eca 109 cells m TOR,HIF-1?,VE-cadherin,MMP-2 and MMP-9.