Effect Of Wind Medicine Regulating PTEN-PI3K-AKT Pathway On Invasion And Metastasis Of Lung Adenocarcinoma

Objective:This research is based on the significant clinical efficacy of wind medicine in the treatment of invasion and metastasis of lung adenocarcinoma,combined with literature analysis and pharmacological research data.It is found that the PTEN-PI3K-AKT pathway closely related to the invasion and metastasis of lung adenocarcinoma;therefore,it was used as the entry point.Mouse Lewis lung cancer cells(LL/2)were used as research objects to explore the effects of wind medicine on lung and adenocarcinoma in vitro and in vivo,and to study its main mechanism of action and its effect on invasion and metastasis of lung adenocarcinoma.This research provides a theoretical basis for further feeding back clinical.Methods:(1)Establish a database of "wind medicine-lung adenocarcinoma invasion and metastasis" through literature review,and screen the top five groups of wind medicine groups with the highest frequency of intervention in lung adenocarcinoma invasion and metastasis.(2)Experiment 1: Cell stable culture,passage,cryopreservation and resuscitation;Cell Migration Experiment and cell invasion experiment were used to study the migration and cell invasion of LL/2 cells by high frequency wind medicine serum.The influence of invasive ability,cell cycle test,soft agar tumorigenesis test and cloning test on cell cycle,and CCK-8 test on cell viability were used to determine the effect of invasive ability on cell cycle.The first three groups of dominant wind medicines with significant screening effect were selected.(3)Experiment 2: Western blot was used to detect the effects of the first three dominant wind medicines on the expression of PTEN,PI3 K and AKT proteins.(4)Experiment 3: Establish a mouse model of Lewis lung cancer cells(LL/2).The low,medium and high dose water decoction made from the dominant wind medicine group selected in Experiment 1 was given orally continuously for 21 days to observe the general situation of each group,the growth of tumors,the weight of tumors,the inhibition rate of tumors and the weight of lungs;HE staining of tumors and lung tissue sections was used to observe the situation of tumor cells and lung metastases by ELISA.The expression of vascular endothelial growth factor in serum was determined by VEGF.(5)Experiment 4: Establish the blood metastasis model of mouse Lewis lung cancer cells(LL/2)and observe the general situation,lung weight,lung volume,lung metastasis and anti-metastasis rate of mice in each group;observe the lung metastasis by HE staining of lung tissue sections;detect the content of serum vascular endothelial growth factor by ELISA.Results:(1)Literature screening: The first five groups of high-frequency wind medicine were: Dilong-Jiangcan group > Wugong-Quanxie group > Jingjie-Fangfeng group > Chuanxiong-Chaihu group > Mahuang-Guizhi group.(2)Experiment 1: Drug-containing serum of Chuanxiong-Chaihu group,Wugong-Quanxie group and Dilong-Jiangcan group could significantly inhibit LL/2 cell invasion,in which Wugong-Quanxie group had more significant ability;drug-containing serum of wind medicine group had no significant difference on LL/2 cell cycle;Drug-containing serum of Wugong-Quanxie group,Dilong-Jiangcan group and Chuanxiong-Chaihu group had the ability to inhibit LL/2 cell.The drug-containing serum of Wugong-Quanxie group,Dilong-Jiangcan group and Chuanxiong-Chaihu group inhibited the cloning ability of LL/2 cells.(3)Experiment 2: In the high-dose group of Wugong-Quanxie,the low-,medium-,and high-dose groups of Dilong-Jiangcan and the high-dose group of Chuanxiong-Chaihu was different to the ratio of PTEN/ACTIN in the blank group(P<0.05),and the medium group of Wugong-Quanxie was more significant(P<0.01).The ratio of PI3K/ACTIN in the low-dose group of Dilong-Jiangcan was different from that of blank group(P<0.05).The medium group of Wugong-Quanxie,the medium dose of Dilong-Jiangcan and the medium-and high-dose group of Chuanxiong-Chaihu had more significant difference(P<0.01);In the Wugong-Quanxie middle dose group and the middle dose group of Chuanxiong-Chaihu of the ratio of AKT/ACTIN was different to the blank group(P<0.05),and the middle dose group of Dilong-Bloodworm and the high dose group of Chuanxiong-Bupleurum had more Significant difference(P<0.01).(4)Experiment 3: The average tumor weight of Chuanxiong-Chaihu high-dose group,Dilong-Jiangcan medium-high dose group,Wugong-Quanxie low-middle-high dose group was different from that of model group(P<0.05);the tumor volume of Dilong-Jiangcan high-dose group was significantly different from that of model group(P<0.05),the Wugong-Quanxie medium-dose group was significantly different(P< 0.01);The lung weight of the low-and medium-dose group of Dilong-Jiangcan and the medium group of Wugong-Quanxie was different from that of the model group(P<0.05);Tumors can be detected by HE in all groups;HE staining of lung tissue except for a small number of cisplatin groups,can be found in metastasis;The group of the drug with a relatively significant tumor inhibition rate was: the medium of Wugong-Quanxie group > the medium of Dilong-Jiangcan group> the high of Chuanxiong-Chaihu group;Serum VEGF levels in the low-middle and high-dose groups of Dilong-Jiangcan,and the low-to-high-dose group in the Wugong-Quanxie were different from those in the model group(P<0.05).Conclusion:Wind medicine can effectively interfere with the invasion and metastasis of lung adenocarcinoma.It can inhibit the invasion and metastasis of lung adenocarcinoma by regulating PTEN-PI3K-AKT signaling pathway,interfering with the expression of VEGF,inhibiting the growth of tumor blood vessels and regulating the heterogeneity of tumor microenvironment.