Effect of silent PRR13 gene on proliferation and apoptosis of colorectal cancerous cells and its possible mechanism
Objective To investigate the effect of silent proline-rich protein 13(PRR13)gene on proliferation and apoptosis of colorectal cancerous cells,and to explore its possible mechanism based on autophagy-associated proteins,immune cell-associated cytokines,and Wnt/β-catenin signaling pathway.Methods(1)The mRNA expression of PPR13 in normal human colorectal epithelial cells(FHC cell),and in human colorectal cancerous cells(HCT116 cell,HT-29 cell,LS174T cell,SW480 cell,SW620 cell,LoVo cell,and HR-8348 cell)were detected,after then human colorectal cancerous cells with the highest expressions were selected to perform follow-up experiment.(2)HR-8348 cell was assigned to control group,empty vector group,PRR13-small interfering RNA(siRNA)group,IWR-1 group,or PRR13-siRNA+IWR-1 group.The control group did not received any intervention,whereas the empty vector group was transfected with negative control siRNA,the PRR13-siRNA group was transfected with PRR13-siRNA,the IWR-1 group received IWR-1 intervention of Wnt/β-catenin signaling pathway inhibitor,and the PRR13-siRNA+IWR-1 group was transfected with PRR13-siRNA and received IWR-1 intervention.PPR13 mRNA expression,cell survival rate,cell apoptosis rate,γ-interferon level,interleukin(IL)-4 level,and expressions of microtubule-associated protein 1 light chain 3 Ⅱ(MAP1LC3Ⅱ),Beclin 1 protein,β-catenin protein,glycogen synthase kinase 3β(GSK-3β)protein,and Wnt11 protein were detected in various groups.Results(1)Except for SW620 cell,PRR13 mRNA expression of the remaining 6 human colorectal cancerous cells was all higher than that of human colorectal epithelial FHC cell,and PRR13 mRNA expression in HR-8348 cell was higher than that in the remaining human colorectal cancerous cells(P<0.05).(2)Compared with the control group and the empty vector group,the remaining 3 groups yielded decreased PRR13 mRNA level,cell survival rate,and IL-4 level,as well as down-regulated expressions of MAP1LC3Ⅱ protein,Beclin 1 protein,β-catenin,GSK-3β protein,Wnt11 protein,and elevated cell apoptosis rate and γ-interferon expression(P<0.05).Compared with the PRR13-siRNA group and the IWR-1 group,the PRR13-siRNA+IWR-1 group exhibited more significant changes of the aforementioned indices(P<0.05),but there was no statistically significant difference in the aforementioned indices between the PRR13-siRNA group and the IWR-1 group(P>0.05).Conclusion Silent PRR13 gene can inhibit colorectal cancerous cell activity,and promote its apoptosis.Its mechanism may by related to regulate Th differentiation,inhibit autophagy-associated protein activity,and activate Wnt/β-catenin signaling pathway.Combining the inhibition of the Wnt/β-catenin signaling pathway with silent PRR13 gene may provide a better anti-colorectal cancer effect.
Colorectal cancerProline-rich protein 13Silent geneAutophagy-associated proteinsHelper T lymphocyteWnt/β-catenin signaling pathwayIn vitro experiment
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