Screening And Constructing A High Metastassis Subline From Lovo Cell

Metastasis of human colorectal carcinoma(HCC),which is the interaction result between tumor and its host, is a complicated process. Tumor cell is the main part of tumor metastasis.Study on biological feature of tumor cell is significant in the research of tumor's genesis and development.Lovo is HCC tumor cell and undifferentiated adenocarcinoma cell.Although it's a strong invasive cell line,its metastasis period is rather long with a high metastasis rate which is up to 40-50% in 4w's metastasis.Lovo cell applied nowadays begins to decline because of the repeated culture of go- down-to future generation in vitro,metastasis rate tends to decrease gradually with a longer metastasis period.This experiment is to construct a model of liver metastasis by means of continous culture of go-down-to future generation in vivo using nude mouse to repeatedly inoculate tumor's cell to its spleen and then separate tumor cell from metastasis field to make some subline tumor cells adapt gradually and show its dorminant growth and become stable after several times inoculation,Simultaneously,we remove those 4 unstable subline to obstain relatively stable ones with a comparatively strong metastasis ability.In addition,this subline possesses a high function of selecting one organ to form metastasis field,Lovo cell chosen by this way can both be cultured in vitro and grow easily in proper organ in vivo.Therefore the lovo cell can be used in various experiment and treatment study for HCC. Main contents of this research include: 1. Apply a continous go-down-to the future generation method of mouse, constructing a liver-metastasized nude mouse's model from spleen inoculation to select strong invasive lovo cell subline through repeated experimental metastasis. 2. From its electrophoresis migration rate. its movement ability its growth curve cell cycline. quantity of DNA and metastasis in vitro to examine lovo cell and analyse its metastasis ability. Main methods and results: 1. Construction of strong invasive lovo cell subline by continous go-down-to future generation in vivo.To found a liver metastasis model by inoculating love cell in nude mouse spleen. 5 2. Make tumor cell's eletrophoresis migration rate examination for strong invasive lovo cell subline and its mother love cell and to study charge properties and density of tumor cell's surface.Result shows the electrophoresis migration rate of lovo cell subline is obviously higher than that of its mother lovo cell. 3. Use MTT method to examine growth curve of strong invasive lovo cell subline and its mother lovo cell in order to study cell's growth speed.The result shows a great difference in growth speed between lovo cell subline and its mother lovo cell. 4. Apply agrose drop method to examine tumor cell's ability to move.Result shows the cell's amount and distance of emigrating agrose drop is obviously higher than that of its mother lovo cell.Apply NIH3T3 cell to test tumor cell's metastasis ability in vitro.The result shows strong invasive lovo cell subline's ability of invasing NIH3T3 cell is obviously higher than that of its mother lovo cell. 5. Apply FCM to examine...