Grb2Inhibition Of Colon Cancer Cells HT29Tumor-associated Signal Transduction

[Objective]To analyze the effect of Grb2shRNA transfected HT29coloncancer cell signaling pathway,and explore the feasibility of theinhibition of Grb2used in the treatment of colorectal cancer.[Method]The Grb2shRNA was transfected to293T cells by shRNA lentiviralvector system, Grb2shRNA lentiviral particles was obtained and infectedthe colon cancer HT29cells.Cell growth inhibition was detected by MTTassay. Grb2mRNA expression was detected by RT-PCR,and Western Blot wasused to measure the altered expression of Grb2,P42/44ERK, phosphorylatedP42/44ERK, serine/threonine protein (Akt), phosphorylated Akt(P-Akt),STAT5and other signaling pathway molecules.[Results]After the Grb2shRNA Lentiviral infection of the colon cancerHT29cells,5infected with shRNA Lentiviral Particles colorectal cancerHT29cell line as the experimental group (ie, cells infected with Grb2shRNA HT29/shGrb2-69, HT29/shGrb2-70, HT29/shGrb2-71, HT29/shGrb2-72,HT29/shGrb2-73),eGFP shRNA lentiviral infection of the colon cancer HT29cells was as a negative control group.72hours after infection by RT-PCR,Western Blot test, we found HT29/shGrb2-69, HT29/shGrb2-73cells, theinhibitory effect of Grb2best. With the control (HT29cells), negativecomparisons, HT29/shGrb2-69, HT29/shGrb2-73cells in24hours of ODvalues were0.176±0.045,0.186±0.013(P<0.001),48hours0.347±0.048,0.382±0.041(P<0.001),72hours0.934±0.038,0.983±0.205(P<0.01).Grb2inhibition can lead to Phospho-p42/44ERK, Akt, Phospho-Aktdecreased, while the ERK, STAT5expression is not affected.[Conclusion]Application of lentivirus vector systems transfection of Grb2shRNA to the HT29cells，leading to Grb2significantly inhibited themRNA and protein levels,and affect HT29cell growth and signaltransduction molecules downregulated.