| Objective:To investigate the effect of regulating TGF-?1 / Smad4 pathway on cervical cancer SiHa cell autophagy genes LC3 and p62.Methods:Cervical cancer SiHa cells with a concentration of 1.5 × 105 / ml in logarithmic growth phase were seeded in a six-well plate,adhered to the wall after 24 hours of routine culture,and starved overnight.The grouping process was as follows: Group A was a group with 0 ng / ml TGF-?1(blank control),Group B was a group with 5 ng / ml TGF-?1,and Group C was a group with 10 ng / ml TGF-?1.Group D was added with 0 ng / ml of SIS3(blank control),group E was added with 5 ng / ml of SIS3,and group F was added with 10 ng / ml of SIS3.Cells were collected after 24 hours.Real-time quantitative PCR and immunoblotting were used to detect the expression of autophagy genes LC3 and p62 at mRNA and protein levels.Results:1.After different concentrations of TGF-?1 acted on SiHa cells for 24 hours,as the concentration of TGF-?1 increased,the expression of LC3 mRNA and protein gradually increased(P <0.05);the expression of p62 mRNA and protein gradually decreased(P<0.05).2.After SiHa cells treated with different concentrations of SIS3 for 24 hours,with the increase of SIS3 concentration,the expression of LC3 mRNA and protein gradually decreased(P <0.05);the expression of p62 mRNA and protein gradually increased(P <0.05).Conclusion:1.The exogenous TGF-?1 / Smad4 pathway activator TGF-?1 can increase the expression of autophagy protein LC3 mRNA and protein,decrease the expression of p62 mRNA and protein,and induce autophagy in cervical cancer SiHa cells;2.The exogenous TGF-?1 / Smad4 pathway inhibitor SIS3 can reduce the expression of autophagy protein LC3 mRNA and protein in cervical cancer SiHa cells,increase the expression of p62 mRNA and protein,and inhibit autophagy in cervical cancer SiHa cells;3.And as the concentrations of TGF-?1 and SIS3 increased,the expression of LC3 and p62 changed,which had a certain concentration dependence. |
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