| Objective:The human epithelial ovarian cancer cell lines of IL-17D genetransfection were subcutaneously inoculated into nude mice respectively, inorder to observe the growth rate of the subcutaneous transplanted tumorvolume; and by measuring the expression of MHC class I-related protein A(MICA)in nude mice, Preliminary to study the factors associated with theeffect of the IL-17D to promote tumor growth, so as to provide a new clue tothe immune escape mechanism, drug-resistant, diagnosis and treatment of theepithelial ovarian cancer.Methods:1The human epithelial ovarian cancer cell lines of SKOV3andOVCAR3with IL-17D gene transfection (SKOV3/17D and OVCAR3/17D)were cultured, as were SKOV3and OVCAR3cells in the blank loadtransfection (SKOV3/neo and OVCAR3/neo).2Establish subcutaneous transplanted model of human epithelial ovarycarcinoma in thirty-two nude mice, they were randomly divided into fourgroups:(1) SKOV3/17D group: subcutaneously inoculated SKOV3cells ofIL-17D gene transfection in nude mice,(2) SKOV3/neo group:subcutaneously inoculated SKOV3cells of blank load transfection in nudemice,(3) OVCAR3/17D group：subcutaneously inoculated OVCAR3cellsof IL-17D gene transfection in nude mice,(4) OVCAR3/neo group：subcutaneously inoculated OVCAR3cells of blank load transfection in nudemice.3The tumor formation rate and the tumor volume were observed, thenthe growth curves of the subcutaneous transplanted tumor were figured.4IL-17D mRNA in tumor tissues of four groups nude mice bearing ovary carcinoma was determined by RT-PCR analysis; The serum levels of sMICAand mMICA expressed on tumor cells was assessed by ELISA and Flowcytometry analysis. Use SPSS13.0statistical analysis software to analyzeexperimental data. P<0.01indicates that the difference had a statisticalsignificance.Results:1The formation rate of transplanted tumor was100%,The growth of tumorin mice inoculted with SKOV3/17D and OVCAR3/17D cells was significantlyfaster compared to that of two corresponding control groups (P<0.01).2IL-17D gene was expressed in each transplanted tumor tissues,theexpression in mice inoculted with SKOV3/IL-17D and OVCAR3/17D cellswas significantly higher compared to that of two corresponding control groups(P<0.01，table1).3Flow cytometry analysis suggsteed that mMICA in each transplantedtumor tissues was expressed, the expressions were showed by meanfluorescence intensity(MFI):3.1The MFI value of MICA in SKOV3/17D group was203.02±3.03,comparing the control group SKOV3/neo285.90±1.76, which showed theexpression of MICA in SKOV3/17D group was obviously lower thanSKOV3/neo group, The difference was statistically significant (P<0.01).3.2The MFI value of MICA in OVCAR3/17D group was177.05±3.69,comparing the control group OVCAR3/neo265.12±2.29, which showed theexpression of MICA in OVCAR3/17D group was obviously lower thanOVCAR3/neo group, The difference was statistically significant (P<0.01).4The serum levels of sMICA(ng/ml) measured by ELISA was expressedin X±s:4.1The serum levels of sMICA in the group of SKOV3/17D andSKOV3/neo were (28.85±0.36) ng/ml and (23.58±0.50) ng/ml respectively,which suggested the level of sMICA in SKOV3/17D group was higher than inSKOV3/neo group(P<0.01).4.2The serum levels of sMICA in the group of OVCAR3/17D and OVCAR3/neo were (34.05±0.42) ng/ml and (27.24±0.51) ng/ml respectively,which suggested the level of sMICA in OVCAR3/17D group was higher thanin OVCAR3/neo group(P<0.01).Conclusion: IL-17D obviously promotes the subcutaneous transplantedtumors of human epithelial ovarian cancer in nude mice, the mechanism mustbe related to the immune kill pathway of NKG2D-MICA, IL-17D candown-regulated MICA on the surface of cancer cells, and enhanced theexpression of sMICA, which leads to the Killing activity of NK cell to cancercells decreases, in this way, tumor can escape from immune surveillance, andacquire tumor immune escape, so we think IL-17D plays an important part inthe progression of human epithelial ovarian cancer. |
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