| [Objective] In this study,based on the significant clinical efficacy of wind drug intervention in the invasion and metastasis of lung adenocarcinoma,combined with literature analysis and pharmacological research data and major pharmacodynamics experiments,the PHD2/HIF-1? signaling pathway,which is closely related to the invasion and metastasis of lung adenocarcinoma,was selected as the starting point for the selection of Lewis lung cancer in mice LL/2 cells were studied to explore the effect of wind medicine on the proliferation and metastasis of Lewis lung adenocarcinoma cells,a nd further by comparing the intervention effect of Centipede Scorpio drugs,Centipede single drug and Scorpio single drug on Lewis lung cancer tumor mice,the related me chanism of its intervention effect was discussed from the angles of angiogenesis of tumor,hypoxia microenvironment and tumor metastasis,which provides a preliminary theoretical and experimental basis for dialectical drug use and reasonable compatibility of lung adenocarcinoma.[Methods](1)Building database: Through literature review to establisha "wind drug-lung adenocarcinoma invasion and metastasis" database,screening anti-lung aden ocarcinoma invasion and metastasis of the highest frequency of the first five groups of wind drugs(1-5 groups);(2)intervention in Lewis lung cancer LL/2 Cell proliferation Transfer experiment: The construction of mouse Lewis lung cancer LL/2 Cell stable culture,Transmission,cryopreservation and resuscitation of cells;detection of cell cycle by PI staining,flow cytometry;Detection of cell proliferation ability and mapping of growth curve by cell proliferation Experiment(CCK8);Detection of the colony for mation ability of LL/2 lung adenocarcinoma cells in apetri dish by clonal formation experiment;The tumor-forming ability of cells was detected by soft agar tumor experiment,the apoptosis ability of lung adenocarcinoma cells was detected by Annexin V/PI flow,and the migration and invasion ability of cells were detected by scratch healing experiment and Transwell experiment.The changes of HIF-I?,PHD-2 and MMP-2 expression in cells were detected by Western blot method to evaluate the effect of wind drug serum on in vitro intervention of lung adenocarcinoma,and a group of wind drugs with the best efficacy was screened.(3)intervention in subcutaneous orthotopic carcinoma of Lewis lung cancer mice: establishment of Lewis lung cancer in mice LL/ 2 cell mouse model,in vitro,the optimum wind drug group was selected to make the water decoction into the stomach,and the model groups were randomly divided into blank control group,model group,cisplatin Group,Centipede-Scorpio Group,Cen tipede Group,Scorpio Group,after the group were given according to the equivalent d ose of human rats,continuous administration for 21 days,detection of the general situ ation of each group,tumor growth curve,tumor weight inhibition rate,lung weight,l ung volume,etc.,the tumor cells and metastases were observed by paraffin tissue slice he staining,and the expression of HIF-I?,PHD-2 and MMP-2 in tissues was detected by Western blot method,and the expression of VEGF in tissue was detected by ELISA method.To verify the effect of group intervention on invasive metastasis of lung adenocarcinoma cells.(4)intervention in the blood transfer experiment of Lewis lung canceand the model groups were randomly divided into blank control group,model group,cisplatin Group,Centipede-Scorpio Group,Centipede Group,Scorpio Group,after the group were given according to the equivalent dose of human rats,continuousad ministration for 21 days,detection of the general situation of each group,tumor growth curve,tumor weight inhibition rate,lung weight,lung volume,etc.,the tumor cells and metastases were observed by paraffin tissue slice he staining,and the expression of HIF-I?,PHD-2 and MMP-2 in tissues was detected by Western blot method,and the expression of VEGF in tissue was detected by ELISA method.To verify the effect of group intervention on invasive metastasis of lung adenocarcinoma cells.[Result](1)Literature Screening: The first 5 groups of high-frequency wind drug sare: Dillon-Silkworm Group > centipede-Scorpio Group > mustard-Windproof group > Chuanxiong-Chaihu Group > ephedra-Guizhi group;(2)intervention in Lewis lung cancer LL/2 Cell proliferation Transfer experiment: Chuanxiong-Chaihu Group,Cetipede-Scorpio Group,Dillon-Silkworm group of drug-containing serumcan be more significantly inhibited LL/2 cells invasion,of which the Centipede-Scorpio group ability was more significant;there was no significant difference in the effect of drug-containing serum on LL/2 cell cycle in each wind group,and the drug-containing serum of the Dillon-Silkworm group,Centipede-Scorpio Group and mustard-Windproof group inhibited the ability of LL/2 cell soft agar tumor,The drug-containing serum of the Dillon-Silkworm group and the Centipede Scorpio group inhibited the cloning ability of LL/2 cells.The drug-containing serum of the Dillon-Silkworm group,Centipede-Scorpio Group and Chuanxiong-Chaihu group had obvious inhibitory effect on the activity of LL/2 cells.(3)The effect of the drug group on the lung weight of the mice: the lung weight of the B group was the least,the difference between the group A and the group A was more significant(P <0.01),and the lung weight of the group C and the E group was more significant(P <0.05).The effect of each drug group on the lung tra nsfer range and the anti-metastasis rate was the difference(P <0.01),and the anti-met astatic rate of group B was the highest in group B.The anti-metastasis rate of each drug group was from high to low: group C> D group> E group.histological observation: histological observation of lung: DDP group> centipede-whole Scorpion formation > centipede formation ? Dilong formation.Each drug group could inhibit the expression of VEGF in different degrees,among which cisplatin group had the most significant effect,followed by centipede-scorpion group and centipede group.(4)intervention of lewis lung cancer mice blood metastasis experiment: the effect of each drug group on the lung weight of mice: the lung weight of group B was the least,and the difference was more significant than that of group A(P < 0.01),C),the lung weight of group E was higher than that of group A(P < 0.05),and the lung weight of group E was lower than that of group A(P < 0.05).There was difference(P < 0.05).Effect of each drug group on lung metastasis and anti-metastasis rate: group B had more significant difference than group A in lung metastasis(P < 0.01),and group B had the highest anti-metastasis rate.The anti-metastasis rate of each drug group was C group > D group > E group from high to low.Histological observation: lung histological observation: DDP group > Centipede-Scorpion Group > centipede group ? whole scorpion group.Each drug group could inhibit the expression of VEGF in different degrees,among which cisplatin group had the most significant effect,followed by centipede-scorpion group and centipede group.[Conclusion] Fengyao has obvious effect on invasion and metastasis of lung ade nocarcinoma.It can regulate PHD2/HIF-1 ? signal pathway and VEGF,MMP-2 content,decrease tumor vascular growth,and regulate tumor microenvironment heterogene ity.Thus inhibit the invasion and metastasis of lung adenocarcinoma. |
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