Investigation On The Effect And Mechanism Of CK2α OnHCT116Colon Cancer Cell Proliferation

ObjectiveCK2α as a research target, RNA interference by observing the growth andproliferation of HCT116cells expressing CK2α expression levels of cellcycle-related proteins circumstances and CyclinH, P53and P21, analysis CK2αon HCT116cell growth and proliferation, and to explore its mechanism.Methods1. According mRNA sequence CK2α designed CK2α-siRNA sequence,HCT116cells cultured in vitro were divided into normal control group, negativecontrol group and CK2α-siRNA group and applied Lipofectamine2000transfection. Inverted phase HCT116cell morphological changes observedunder the microscope after interference CK2α. RNA blotting method to detectthe use of Western interference24h,48h,72h,96h protein levels at differenttime points CK2α, and thus interfere with the analysis of the effect of CK2α-siRNA.2. MTT assay were used to detect RNA interference24h,48h,72h,96h atdifferent time points HCT116cell proliferation.3. Flow cytometry was used to detect the RNA interference72h HCT116cell cycle phase distribution. 4. Western blotting was used to detect the use of RNA interference after72h HCT116cells CK2α, CyclinH, P53and P21protein expression levels.5. HCT116cells were transfected with MHA and MHA-CK2α, the use ofco-immunoprecipitation assay CK2α interaction with CyclinH.Results1. After CK2α-siRNA interference HCT116cell morphology occurredretraction, cell density decreased.2. CK2α-siRNA interference effects, compared with the negative controlgroup, between24h transfection to72h, HCT116cells CK2α protein levelsgradually decreased CK2α-siRNA,72h interference effect is the mostsignificant, but the expression of transfected96hCK2α level trend of recovery.3. MTT assay, compared with the negative control group transfected48and72h, CK2α-siRNA group was significantly reduced cell viability (P <0.01);transfected24h and96h no significant changes in cell viability (P>0.05).4. Flow cytometric analysis, compared with the negative control group,significantly reduced (P <0.01) the proportion of CK2α-siRNA group aftertransfection72h S phase cells in the cell cycle, G1phase cells was significantlyincreased (P <0.01) cells remain in the G1phase.5. Compared with the negative control group, was significantly reducedCK2α-siRNA group after transfection72h expressing CK2α and cyclin CyclinHof (P <0.01); P53protein expression levels had no significant change (P>0.05);P21protein expression levels were significantly up higher (P <0.01)..6. Co-IP experiments confirmed the presence of CK2α and CyclinHinteractions in vivo.ConclusionsInterference CK2α expression was significantly inhibited the proliferation ofHCT116cells, which may be by inhibiting the expression of cyclin CyclinH andraised P21expression, thereby blocking cell cycle, inhibition of cell proliferation.