Mechanism Analysis How Interleukin-17D Up-regulate The Production Of SMICA

Objective: To observe the effect of rhIL-17 A on cell growth in SKOV3 cells by treating the culture cells with rh IL-17D;and by measuring the expression of MHC class I-related protein A(MICA)in the culture supernatants,Preliminary to study the factors associated with the effect of the IL-17 D to promote tumor growth and to discuss the role of IL-17 D in regulating the production of sMICA,so as to provide a new clue to the immune escape mechanism,diagnosis and treatment of the epithelial ovarian cancer.Methods:1 The human epithelial ovarian cancer cell lines of SKOV3 were cultured.2 SKOV3 cells were cultured with 0,10,50 and 100 ng/mL of recombinant human IL-17D(RhIL-17D)for 24 h,and the expression of sMICA was detected by ELISA.the growth condition of SKOV3 cells was observed.3 Real-time quantitative polymerase chain reaction(qRT-PCR)were used to detect the expression of MMP-9 after SKOV3 cells were incubated with 50 ng/mL Rh IL-17 D for 24 h.4 Western blot was used to detect the expression of MMP-9,p65 and phosphorylation-p65(P-p65).5 When helenalin,a NF-Kb inhibitor,was added to SKOV3 cells medium before rhIL-17 D treatment,MMP9 protein expression was significantly decreased(P,0.05,Independent Student's t-test).Results: The effect of RhIL-17 D on growth in SKOV3 cells After SKOV3 cells cultured with 0,10,50 and 100 ng/m L of recombinant human IL-17D(RhIL-17D)for 24 h,the growth condition of SKOV3 cells was observed,we find 50 ng/mL RhIL-17 D group is greater than other groups;2?The serum levels of sMICA in the group of0,10,50,100ng/mLRhIL-17 D were(162.09±11.28)pg/ml,(176.91±6.04)pg/ml,(165.91±8.12)pg/ml,(166.64±10.00)pg/ml respectively,which suggested the level of sMICA in50ng/mLRhIL-17 D group was higher than in other groups(P<0.05);3?The expressions of MMP9 mRNA in control groups were 170 times as much as 50ng/mLRhIL-17 D groups,compared with control group,significant differences were found in each groups(P<0.05);Addition of RhIL-17 Dresulted in a significant increase in the production of MMP9 in SKOV3 cells;4?RhIL-17 D also increased the MMP-9 and P-p65 protein levels.When helenalin,a NF-Kb inhibitor,was added to SKOV3 cells medium before rhIL-17 D treatment,MMP9 protein expression was significantly decreased.Conclusion:IL-17 D obviously promotes the growth of SKOV3 cells the mechanism must be related to the immune kill pathway of NKG2D-MICA,IL-17 D may up-regulate the production of sMICA by activating the nuclear factor-kappa B(NF-?B)-MMP-9 signal pathway.