Studay On The Mechanism Of Antiphospholipid Antibody Promotes Invasion And Metastasis In Non-small Cell Lung Cancer

Objective:To study the expression of antiphospholipid antibodies(a-PLs),anticardiolipin antibodies(ACA)and anti-?2glycoprotein?(anti-?2GP?)in serum of non-small cell lung cancer,Count the microvessel density in non-small cell lung cancer tissues,explore the correlation between serum anti-phospholipid antibodies and infiltration,metastasis and vascular density of lung cancer.Dteceting the effect of APS-derived aPL~+(IgG)on the expression of CSF-1,TF and VEGF in lung cancer.Then observing the effect of aPL~+(IgG)on the growth and migration of lung cancer cells.To provide a new idea for the mechanism of invasion and metastasis of non-small cell lung cancer.Methods:(1)Chemiluminescence was used to detect the expression of anticardiolipin antibody(ACA-IgG,IgM)and anti-?2glycoprotein?(anti-?2 GP?-IgG,IgM)in lung cancer group and healthy control group,The immunohistochemical method was used to detect the density of blood vessels in lung cancer tissues.(2)The total IgG of patients with antiphospholipid syndrome was extracted by affinity chromatography,the purity of IgG extracted was detected by SDS-PAGE.(3)Flow cytometry and immunofluorescence were used to detect the phosphatidylserine exposure of lung cancer A549,H1299 and H460 cells.(4)RT-PCR was used to detect the expression of CSF-1,TF and VEGF levels in lung cancer A549 cells treated with aPL~+(IgG)at different time points.(5)CCK-8 was used to detect the Proliferative ability of A549 cells treated with different concentrations of control and aPL~+(IgG),Cell cloning experiments were used to further verify cell proliferation(6)Transwell experiments were performed to detect the impact of aPL~+(Ig G)on the invasiveness of lung cancer A549.Results:(1)Compared with the control group,the expression of anticardiolipin antibody(ACA-IgG,IgM)and anti-?2glycoprotein?(anti-?2GP?-IgG,IgM)in non-small cell lung cancer was higher,the difference was statistically significant(p<0.05);Compared with the group without lymph node metastasis,the expression of anticardiolipin antibody(ACA-IgG)and anti-?2glycoprotein I(anti-?2 GP?-IgG)in lymph node metastasis groups was higher,the difference was statistically significant(p<0.05).Compared with the in situ carcinoma group,the expression of anticardiolipin antibody(ACA-IgG,IgM)and anti-?2glycoprotein?(anti-?2GP?-IgG,IgM)in carcinoma was higher,the difference was statistically significant.(p<0.01).(2)Microvessel density in non-small cell lung cancer increased with the progress of lung cancer,stage?was significantly higher than stage?and stage?,lymph node metastasis groups was higher than that of no metastasis groups.(P<0.05).(3)The anticardiolipin antibody(ACA-IgG)and anti-?2 glycoprotein?(anti-?2GP?-IgG)were significantly correlated with the microvessel density in lung cancer(P<0.0001 R=0.2855,P=0.0001 R=0.2264).(4)Phosphatidylserine was exposured on the surface of lung cancer cells A549,H1299,H460.(5)Compared with the control group,the expression of CSF-1,TF and VEGF in aPL~+(IgG)groups were up-regulated,the difference was statistically significant(p<0.01).(6)Compared with the control group,the proliferation and invasion ability of aPL~+(IgG)-treated groups increased significantly(p<0.05).Conclusions:(1)Compared with the control group,the concentration of antiphospholipid antibody in serum of non small cell lung cancer patients is higher.(2)There is a significant correlation between antiphospholipid antibodies and the invasion,metastasis,vascular density of non-small cell lung cancer.antiphospholipid antibodies may promote the progress of lung cancer by promoting the generation of lung cancer blood vessels.(3)Purified IgG from serum of patients with APS can promote the expression of CSF-1,TF and VEGF in lung cancer A549 cells.It is possible to promote the malignant transformation of lung cancer by upregulating the expression of CSF-1,TF and VEGF.(4)aPL~+(IgG)can promote lung cancer A549 cell proliferation and migration...