The Pilot Research Of The Relationship Between Vasculogenic Mimicry Of Lung Cancer And Apoptosis

Objective:Through the three-dimensional cultures of lung cancer cells in vitro to explore if they can form capillary-like structures(CLS), Also is to see whether it has the ability of forming vasculogenic mimicy(VM). Then use VEGF monoantibody to intervene the NCI-H446.And Z-VAD-FMK and caspase-3 shRNA (h) Lentiviral Particles to intervene the expression of caspase-3 in NCI-H446. Through these to see the change of forming CLS and investigate the relationship between apoptosis and VM. And discuss the clinical significance of VM and anti-apoptotic as new lung cancer therapeutic targets.Content:First, to culture NCI-H446 and H1299 in two- dimensional in vitro, and change to three-dimensional cultures until 4-7 generation. And to check the difference between the two types of lung cancer. The use monoclonal anti-VEGF antibody, and Z-VAD-FMK to intervene the three-dimensional culture of NCI-H446. Through these to see the change of forming CLS among different groups. Furthermore using caspase-3 shRNA (h) Lentiviral Particles to intervene the expression of caspase-3 in NCI-H446 and check the change of forming CLS.Method:1. Three-dimensional culture of theNCI-H446 and H1299 in vitro.2. Using monoclonal anti-VEGF antibody to intervene the three-dimensional culture of NCI-H446. Through these to see the change of forming CLS among different groups.3. Using Z-VAD-FMK to intervene the three-dimensional culture of NCI-H446. Through these to see the change of forming CLS among different groups.4. Using caspase-3 shRNA (h) Lentiviral Particles to intervene the expression of caspase-3 in NCI-H446 and check the change of forming CLS. Using RT-PCR and western-blot to see the result of intervene.Results:1. The lung cancer cells can form CLS and have the ability of forming VM. And we found that NCI-H446 had stronger ability of forming CLS than H1299. 2. NCI-H446 can express VEGF protein and NCI-H446 can not form CLS when added VEGF monoantibody in the early stage, but only affected a little when added after forming CLS.3. The results of three-dimensional culture of the NCI-H446 interveneed by ZVAD-FMK showed:NCI-H446 can not form CLS when added in the early stage, but had no affect when added after forming CLS.4. The results of three-dimensional culture of the NCI-H446 interveneed by caspase-3 shRNA (h) Lentiviral Particles showed:the experimental group can not form CLS but the control group can form CLS.Conclusion:1. The lung cancer cells can form CLS and have the ability of forming VM. And NCI-H446 had stronger ability of forming CLS than H1299.2. NCI-H446 can express the protein of VEGF, and VEGF can stimulate the formation of CLS and monoclonal anti VEGF antibody can affect forming CLS.3. Apopsis can stimulate the formation of VM. And the inhibitor of apoptosis can affect forming of CLS in the early stage.4. For the VM positive patients, targeted therapy should consider the vascular endothelial cells and the tumor cells. If the apoptosis was approved as an initiators of the formation of VM in lung cancer, then the anti-apoptosis medicine will be expected to become targeted therapy as EGFR-TKI.