Study On Regulating TLR3and Its Signal Transduction Pathway To Inhibit Cervical Carcinoma In Vitro

ObjectiveTo explore TLR3/PI3K signal transduction pathway’s role in cervical cancer and the relationship between HPV18E6/E7and TLR3/PI3K signal pathway. To provide theoretical base for finding cervical cancer pathogenesis and exploring new ways of treatment of cervical cancer.Methods1RT-PCR and Western blotting were used to detect Hela cells’ TLR3and PI3K in the mRNA and protein level after polyI:C and LY294002’s irritating to Hela cells.2Use the HPV18E6E7siRNA carried by lentivirus which was already synthetized by our research team, put the recombinant DNA into Hela cells by the way of virus infection. RT-PCR and Western blotting way were used to detect Hela cells’ TLR3and PI3K in the mRNA and protein level.3MTT assay was used to analyze the growth of the cells.Results1In the experiment of polyI:C and LY294002irritating Hela cells,in polyI:C group,PI3K and TLR3’s mRNA expression level were71.34±0.70and69.69±4.09.In contrast group, PI3K and TLR3’s mRNA expression level were43.98±1.60and35.19±8.09.PolyI:C group is higher than contrast group and there was significant difference between the two groups(P<0.001;P<0.001).In polyI:C group,PI3K and TLR3’s protein expression level were89.98±3.10and85.77±3.79,while in contrast group those were80.18±437and72.18±1.66. PolyLC group is higher than contrast group and there was significant difference between the two groups(P=0.009;P=0.001).In LY294002group, PI3K and TLR3’s mRNA expression level were28.63±2.38and22.69±3.82,which were lower than those in contrast group and there was significant difference between the two groups(P<0.001;P=0.025).In LY294002group, PI3K and TLR3’s protein expression level were70.42±1.03and63.26±1.53,which were lower than those in contrast group and there was significant difference between the two groups(P=0.009;P=0.005).Cell proliferation rate was20.54±4.21(%) in polyI:C group while cell growth inhibition rate was61.92±5.49(%) in LY294002group. 2In experiment of lentivirus transfecting Hela cells,in transfection group, HPV18E6、 TLR3and PI3K’s mRNA expression level were(0.1360±0.0140)、(0.2367±0.0321) and (0.2013±0.0461), and those in contrast group were (0.392±0.036)、(0.4700±0.0346) and (0.3767±0.0321). Transfection group is lower than contrast group and there was significant difference between the two groups (P<0.001;P<0.001;P=0.005).HPV18E6、TLR3and PI3K’s protein expression level were (0.1670±0.0290)、(0.6834±0.0173)、(0.5670±0.0440), and those in contrast group were (0.2860±0.0370)、(0.8743±0.0263)、(0.7599±0.0403).Transfection group is lower than contrast group and there was significant difference between the two groups (P=0.005;P=0.002;P=0.005). Cell growth inhibition rate was (39.6±18.0)%in transfection group. Cell growth inhibition rate was (3.1±13.3)%in contrast group. There was significant difference between the two groups(P=0.047).Conclusions1.TLR3/PI3K signal pathway played an important role in the growth of cervical cancer cells. LY294002can reduce the expression of TLR3、PI3K to inhibit Hela cells’ growth.2. TLR3/PI3K signal pathway played an important role in trasformming cervical cancer by HPV18E6/E7and cervical cancer cells’growth. Suppressing the expression of HPV18E6/E7can inhibit expression of TLR3/PI3K in mRNA and protein level,then Hela cells’growth were restrained.3.Medicine and gene way can also down regulate TLR3/PI3K singnal pathway and restrain Hela cells’growth,which can supply an target spot for treatment of cervical cacinoma.