The Effect Of Buyang Huanwu Decoction On Lung Cancer A549 Cells Under Hypoxic Microenvironment And The Regulatory Mechanism Of Related Protein Gene Expression

Objective: This study aims to establish the hypoxia model of lung cancer A549 cells and explore the effect of Buyang Huanwu Decoction on hypoxia microenvironment of lung cancer and the regulation mechanism of related protein gene expression.Methods: Establish a hypoxia model of lung cancer A549 cells,divided into5 groups: A.Blank control group(normal oxygen group,A549 cells are routinely cultured in an incubator containing 21% O2,5% CO2,and 74% N2)B.Hypoxia control group(hypoxia group,A549 cells are cultured in an incubator containing 1% O2,5% CO2,and 94% N2)C.Low-dose Buyang Huanwu Decoction group(hypoxia group)(15mg/ m L)D.Medium-dose Buyang Huanwu Decoction group(hypoxic group)(30 mg/m L)E.High-dose Buyang Huanwu Decoction group(hypoxic group)(60 mg/m L);each group has 3 repetitions test.Perform flow cytometry to detect cell proliferation,transwell test to detect cell invasion,cell scratch test to detect cell migration,A549 cell culture solution to treat HUVEC and observe its angiogenesis;Western Blot method to detect lung cancer A549 cells HIF-1α,p53,VEGF,OPN,PDGF-B protein expression,RT-q PCR method for real-time fluorescence quantitative detection of HIF-1α,P53,VEGF,OPN,PDGF-B.Results: Compared with the normoxic group,the expression of HIF-1α,VEGF,OPN,and PDGF-B in the hypoxia control group increased,while the expression of p53 gene decreased.Compared with the hypoxia control group,the expression of HIF-1α,VEGF,OPN,PDGF-B in each group(C,D,E)of Buyang Huanwu Decoction decreased,while the protein expression of p53 gene increased;Buyang Huanwu Decoction Comparing the groups with different concentrations,the expression of HIF-1α,VEGF,OPN,PDGF-B in Buyang Huanwu Decoction 15mg/ml group,30mg/ml group and 60mg/ml group decreased sequentially,while the expression of p53 gene increased sequentially.The results suggest that the expressions of HIF-1α,VEGF,OPN,and PDGF-B in lung cancer A549 cells under hypoxic environment are up-regulated compared with normoxic environment,while the expression of p53 is inhibited.Buyang Huanwu Decoction can reduce the expression of HIF-1α and VEGF,OPN,PDGF-B in lung cancer A549 cells under hypoxia.The inhibitory effect is positively correlated with the concentration of Buyang Huanwu Decoction,and it can up-regulate the expression of p53 gene and activate the effect.It is positively correlated with the concentration of Buyang Huanwu Decoction.Conclusion: Buyang Huanwu Decoction can improve lung cancer A549 hypoxic microenvironment,promote tumor cell apoptosis,and inhibit angiogenesis by inhibiting the expression of HIF-1α and VEGF,OPN,PDGF-B in lung cancer A549 cells,and up-regulating the expression of p53 gene.Reduce tumor invasion and migration,so as to achieve anti-cancer effect.