Related Study And Expression Of Vasculogenic Mimicry (VM) In Tumors Of Digestive System Under Three-dimensional Cell Culture

Objectives of the Experiment1. To build vitro models of a pancreatic cancer cell line SW1990and a gastriccancer cell line SGC-7901with three-dimensional cell culture technology.2. To investigate ability to form vascular mimicry (Vasculogenic mimicry, VM) intumors of digestive system.Methods of the ExperimentIn vitro, to build three-dimensional culture tumor models of pancreatic cancer cellsSW1990and the control group gastric cancer cell line SGC-7901as follows: in2Dculture system, that is, the traditional cell culture model, pancreatic cancer cell lineSW1990and gastric cancer cell line SGC-7901were inoculated in complete solution(90%RPMI1640medium+10%fetal calf serum+antibiotic) to cultivate cells in tissueculture flask and then flasks were shifted into the37°C,5%CO2humidified incubator.Complete solution (90%RPMI1640medium+10%fetal calf serum+antibiotic) wasreplaced every24hours or48hours. Subculture was followed when integration of thecells reached70%-80%.In three-dimensional culture system, the96-well plate was placed on the ice, onwhich Matrigel glue was combined with medium suspension (The cells were mixed at a1:1ratio with precooled serum-free RPMI1640medium) of1×103gastric cancer cellline SGC-7901and pancreatic cancer cells SW1990in the logarithmic growth phase.Cell suspension was made,70μl of which was seeded in96-well plates. Then96-wellplate was quickly placed in a37°C incubator for2minutes. After gelling the mixture,100μl complete medium was added into each well and covered, then placed in37°C,5%CO2humidified incubator. Complete medium (90%RPMI1640medium+10%fetalcalf serum+antibiotic) in the96-well plate was replace every24hours for propapation.After24hours,48hours,72hours,96hours respectively, the state of cell growth and blood vessel growth mimicry (VM) formation were observed and recorded by camerasrandomly under an inverted microscope.Results of the Experiment1.Epithelioid differentiated gastric adenocarcinoma cell line SGC-7901in thetwo-dimensional cell culture, grew adherently after2-4hours and was followed bysubculture within24-72h after inoculation, and when confluence of the cells reached90%, the experiment could be carried out. In matrigel, gastric cancer cell line SGC-7901suspended in a hydrogel after inoculation were round and cells proliferation and a largenumber of incomplete lumen formation could be seen after24-48hours; grid-like VMwas visible after72-96hours. In the model formed by the three-dimensional cellculture of the epithelioid differentiated gastric adenocarcinoma cell line SGC-7901,formation of a network-like vascular mimicry (VM) could be seen after24hours underthe observation of an inverted microscope.2. In the two-dimensional cell cultures, pancreatic cancer cells SW1900grewadherently within4-8hours and cells were in fusiform or polygon, and then followed bysubculture after72-96hours. When confluence of the cells reached90%, experimentscould be carried out. In matrigel, SW1990pancreatic cancer cells suspended in ahydrogel after inoculation were circular; the cell division and growth were seen after24-48hours and no VM was observed. The cells were still growing after72-76hours ata slower pace without formation of VE.agglomerate growth could not be observed inpancreatic cancer cells SW1900of three-dimensional cell culture in vitro under opticalinverted microscope24-48hours and the same phenomenon happened after72-96hours.Formation of vasculogenic mimicry (VM) was not observed in pancreatic cancer cellsSW1990of3D cell culture.Conclusion of the Experiment1. The differences exist in the VM expression of different tumors of digestivesystem.2. The VM may play a role in the occurrence, development, invasion, metastasisand other malignant behaviors of gastric cancer.3. There is no significant correlation between VM with the occurrence,development, invasion, metastasis and other biological behaviors of pancreatic cancer.