Analysis Of Silent Twist Gene And Relative Molecules Of PT3K/Akt/Bad Signal Pathway In Cervical Cancer Caski Cell Line

Objective: Exploring the Silent Twist gene and the relation of Taxol chemotherapysensitivity in the Caski cells of cervical cancer,and analysis the relation ofPI3K/Akt/Bad signal pathways related proteins.Methods: The Twist gene to lowed expression vector sequence was analyzed withBioinformatics methods.The Twist gene of restructuring of the carrier interference(Twist-shRNA) was Built,and then it was transferred stably by liposome to Caskicells of human cervical cancer to culure a stable Twist gene to lowed expression ofcell line (Caski-Twist-shRNA) and blank vector transfection group (Caski-shRNA--NC group).Twist mRNA of each group is expressed after RT-PCR’s verification andtransfection, through treating Paclitaxe with different concentrations, MTT tests thecell proliferation ability, Western blot tests the expression of PI3k,Akt and Badprotein.Results: Twist-shRNA was transferred to Caski cells by liposome, and culured astably cell line to lowed expression Twist gene (Caski-Twist-shRNA) and blankvector transfection group (Caski-shRNA-NC).After the expression of silent Twistgene, the Twist mRNA expression level of Caski-Twist-shRNA group compared thenegative control group (Caski shRNA-NC) and normal control group(Caski)decreased obviously,which is of significanct difference (p <0.05).Groups of cells areprocessed with different concentration. Compared with the negative control group(Caski-shRNA-NC+PTX) and the blank control group (Caski), the proliferationinhibition rate in experimental group (Caski-Twist-shRNA+PTX) increases obviously(p <0.05). IC50alue concentration in experimental group is lower than the negativecontrol group and the blank control group (p <0.05),and the optimal concentrations ofPaclitaxel for1μmol/L will be selected.Western blot results showed: Compared withCaski-shRNA-NC+PTX group, Caski+PTX group and Caksi cell group, PI3k andAkt protein expression level in Caski-Twist-shRNA+PTX group is obviously lower(p<0.05), while Bad protein expression in the Caski-Twist-shRNA+PTX groupincreases obviously (p <0.05) compared with Caski-shRNA-NC+PTX group,Caski+PTX group and Caksi cell group.Conclusion:The lowered Twist gene increases the Paclitaxel chemotherapy’ sensitivity in cervical cancer Caski, and lead to the expression of lowed PI3k, Akt andraised Bad.