The Influence Of Ovarian Cancer Specific Targeting Peptide On The Protein Expression Of Ovarian Cancer Cell A2780 In IKK/NF-κB Pathway

Objective:Our group has successfully screened and synthesized the varian cancer specific targeting peptide(OSTP) for the first time.It has certified that OSTP can promote ovarian cancer A2780 cell apoptosis and Sensitied the effect of promote ovarian cancer A2780 cell apoptosis of paclitaxel by in vitro experiments.The purpose of this experiment is to confirm OSTP promote ovarian cancer cell and sensitied the effect of promote ovarian cancer apoptosis of paclitaxel by IKK/NF- κ B pathway.The results of this experiment will provide the experimental basis for clinical ovarian cancer therapy of OSTP.Method: 1. Synthesised OSTP 2. Culturing ovarian cancer A2780 cell lines in vitro,randomly divided the ovarian cancer A2780 cell lines in logarithmic phase into 1640 blank control group, DMSO group, OSTP group and The OSTP taxol group.Treat cells for 24 h,then extract the cellular proteins.Testing the expression of IKKβ, Phosphorylation of IKK(p- IKKβ), NF-κB(p65),Phosphorylation of NF-κB(p- NF-κB) by Western Blot. 3. Culturing ovarian cancer A2780 cell lines in vitro,randomly divided the ovarian cancer A2780 cell lines in logarithmic phase into 1640 blank control group, OSTP 15 min group, OSTP 30 min group, OSTP 1h group, OSTP 3h group, OSTP 6h group and OSTP 12 h group. Extract the cellular proteins respectively in 15min、30min、1h、3h、6h and 12 h.Testing the expression of IKKβ, Phosphorylation of IKK(p-IKKβ), NF-κB(p65), Phosphorylation of NF-κB(p- NF-κB) by Western Blot.Result: 1. The successful synthesis of OSTP. 2. Compared with control group, the expression of IKKβ in OSTP15 min group, 30 minutes group, 1 h group, 3 h group, 6 h group, 12 h group have no significant difference(p>0.05). The expression of pIKKβ(Tyr188)is lower than control group in OSTP15 min group, 30 minutes group, 12 h group(p<0.01); However the expression of p- IKKβ(Tyr188)in OSTP1 h group,3h group and 6h group is higher than control group(p<0.01);The lowest IKKβ(Tyr188)expression is OSTP 15 min group,and the highest IKKβ(Tyr188)expression is OSTP3 h group. 3. The expression of NF-κB(p65) in OSTP 1h group, 3h group, 6 h group, 12 h group are lower than control group(p<0.01).After OSTP treated for 1h,the expression of NF- κ B( p65) Gradually reduce(p<0.01). The expression of NF-κB(p65 Ser311)phosphorylated protein in OSTP 15 min group, 30 min group, 1h group,3h group,6h group,12 h group are lower than control group(p<0.01). The lowest NF-κB(p65Ser311)phosphorylated protein expression is OSTP 30 min group. 4. The expression of IKKβ in taxol group is lower than control group. Compared with control group,the expression of IKKβ in OSTP group have no significant difference(p>0.05).Compared with taxol group,the expression of IKKβ in combined drugs group have no significant difference(p>0.05). The expression of IKKβ( Tyr188)phosphorylated protein in OSTP group and taxol group is lower than control group(p<0.01). The expression of IKKβ(Tyr188)phosphorylated protein in combined drugs group is lower than OSTP group and taxol group(p<0.01). 5. The expression of NF-κB(p65)in OSTP group and taxol group are lower than control group(p<0.01). The expression of NF-κB(p65)in combined drugs group is lower than OSTP group and taxol group(p<0.01). The expression of NF-κB(p65 Ser311)phosphorylated protein in OSTP group and taxol group is lower than control group(p<0.01). The expression of NF-κB(p65 Ser311)phosphorylated protein in combined drugs group is lower than OSTP group and taxol group(p<0.01).Conclusion: 1. OSTP have the ablity to promote Ovarian cancer A2780 cell apoptosis,the mechanism of it may be related to the reduction of IKKβ(Tyr188)phosphorylated protein、NF-κB(p65)、NF-κB(p65Ser311)phosphorylated protein in IKK/NF-κB pathway. 2. OSTP have the ability to sensitization paclitaxel induced ovarian cancer A2780 cell apoptosis,the mechanism of it may be related to the reduction of IKKβ(Tyr188)phosphorylated protein、NF-κB(p65)、NF-κB(p65 Ser311)phosphorylated protein in IKK/NF-κB pathway.