The Effect Of Knockdown ILK By ShRNA On The Proliferation, Migration And Invasion Of Human Glioma Cell U251

Objective: To investigate the change of proliferation, migration, invasion and tumorigenesis of human brain glioma cells U251 after knockdown ILK by sh RNA interference.Methods:1)U251 were transfected with the specific sh RNA vector and stable tranfected cells were established. Western-blot,RT-PCR are separately performed to detect the expression of ILK m RNA, ILK in U251 cell line and in stable tranfected cells strain samples respectively. ILK expression was significantly reduced.2)the effect of sh RNA-ILK on cell proliferative capacity;(1) CCK-8 is used to detect Changes in cell proliferation rate after transfection.(2) Flow cytometry is used to detect the change of cell cycle before and after transfection(3) Immunofluorescence is used to detect the expression of Ki67. Western-blot,RT-PCR are separately performed to detect the expression of Cyclin D1,Cyclin D1 m RNA in U251 cell line and in stable tranfected cells strain.3)the effect of sh RNA-ILK on cell Cell migration, invasion capability;(1) Transwell chamber is used to detect the change of cell migration, invasion capability before and after transfection;(2) Immunofluorescence is used to detect the expression of E-cadherin. Western-blot,RT-PCR are separately performed to detect the expression of E-cadherin and Snail,E-cadherin m RNA and Snail m RNA in U251 cell line and in stable tranfected cells strain.4)The establishment and observation of the nude mice model of U251 human glioma xenografts.Results:(1) The expression of ILK and ILK m RNA decreased significantly(2) The proliferation, migration and invasion capability of U251 after knockdown ILK is reduced; the indicators related to proliferation, migration and invasion has changed significantly; these indicators are significantly different from those of the control group(P<0.05).(3) The tumorigenesis capability in nude mice of U251 knockdown ILK by sh RNA is significantly reduced.Conclusion: The proliferation, migration, invasion and tumorigenesis capability of U251 after knockdown ILK by sh RNA is significantly reduced. Therefore, ILK might be a critical gene for the inhibition of glioma.