CAJ | 학술논문

目的探讨B 16F10-ESAT-6-gpi/IL-21瘤苗抑制小鼠黑色素瘤肺转移的效应和机制.方法将12只C57BL/6小鼠,分为瘤苗免疫组(给予B16F1 0-ESAT-6-gpi/IL-21瘤苗免疫)和对照组(未给予瘤苗免疫).B 16F10-ESAT-6-gpi/IL-21瘤苗免疫小鼠后,经尾静脉注射B16F10细胞,观察肺转移情况;采用细胞毒实验分析瘤苗免疫小鼠的细胞毒性T淋巴细胞活性;酶联免疫吸附试验检测免疫小鼠的血清γ-干扰素(IFN-γ)水平;逆转录聚合酶链反应方法检测瘤组织中转化生长因子β2(TGF-β2)、锌指增强子结合蛋白1(ZEB1)、丝裂原活化蛋白激酶4和β-肌动蛋白的表达;免疫荧光检测瘤组织中E-钙黏蛋白和N-钙黏蛋白的表达.结果瘤苗免疫鼠肺部肿瘤的结节数明显少于对照组.对照组和瘤苗免疫组小鼠肺脏重量分别为(406.3±27.12)mg和(285.8±19.01)mg,差异有统计学意义(P-0.005 4).瘤苗免疫组小鼠CD8+T细胞对靶细胞的杀伤率[(42.62±3.465)％]明显高于对照组[(22.29±1.804)％],差异有统计学意义(P =0.003 2).瘤苗免疫组和对照组小鼠血清中IFN-γ水平分别为(55.200±7.173) pg/ml和(6.435±1.339) pg/ml,差异有统计学竟义(P=0.0003).免疫鼠瘤组织TGF-β2、ZEB1、N-钙黏蛋白表达水平下降,而E-钙黏蛋白表达水平上升.结论 B16F10-ESAT-6-gpi/IL-21瘤苗免疫能抑制小鼠黑色素瘤肺转移,这与CD8+T细胞为主的细胞免疫应答有关,与瘤苗诱导高水平IFN-γ影响肿瘤组织上皮间质转换有关.
목적 탐토B 16F10-ESAT-6-gpi/IL-21류묘억제소서흑색소류폐전이적효응화궤제.방법 장12지C57BL/6소서,분위류묘면역조(급여B16F1 0-ESAT-6-gpi/IL-21류묘면역)화대조조(미급여류묘면역).B 16F10-ESAT-6-gpi/IL-21류묘면역소서후,경미정맥주사B16F10세포,관찰폐전이정황;채용세포독실험분석류묘면역소서적세포독성T림파세포활성;매련면역흡부시험검측면역소서적혈청γ-간우소(IFN-γ)수평;역전록취합매련반응방법검측류조직중전화생장인자β2(TGF-β2)、자지증강자결합단백1(ZEB1)、사렬원활화단백격매4화β-기동단백적표체;면역형광검측류조직중E-개점단백화N-개점단백적표체.결과 류묘면역서폐부종류적결절수명현소우대조조.대조조화류묘면역조소서폐장중량분별위(406.3±27.12)mg화(285.8±19.01)mg,차이유통계학의의(P-0.005 4).류묘면역조소서CD8+T세포대파세포적살상솔[(42.62±3.465)％]명현고우대조조[(22.29±1.804)％],차이유통계학의의(P =0.003 2).류묘면역조화대조조소서혈청중IFN-γ수평분별위(55.200±7.173) pg/ml화(6.435±1.339) pg/ml,차이유통계학경의(P=0.0003).면역서류조직TGF-β2、ZEB1、N-개점단백표체수평하강,이E-개점단백표체수평상승.결론 B16F10-ESAT-6-gpi/IL-21류묘면역능억제소서흑색소류폐전이,저여CD8+T세포위주적세포면역응답유관,여류묘유도고수평IFN-γ영향종류조직상피간질전환유관.
Objective To investigate the effect and mechanism of B16F10-ESAT-6-gpi/IL-21 tumor cell vaccine on pulmonary metastasis in mouse model of melanoma.Methods Twelve 8-week old female C57BL/6 mice were used in this study.The mice were injected with wild-type B16F10 cells through tail vein after immunization with B16F10-ESAT-6-gpi/IL-21 tumor cell vaccine,and the pulmonary metastasis was observed.The CD4 + and CD8 + T cells were isolated by magnetic activated cell sorting,and then used for the detection of CFSE/7-AAD cytotoxicity by flow cytometry.Serum from the mice immunized with tumorcell vaccine was used to detect IFN-γ expression by ELISA.The expression of TGF-β2,ZEB1,E-cadherin,and N-cadherin of tumor tissues was detected by RT-PCR and immunofluorescence,respectively.Results The mice vaccinated with B16F10-ESAT-6-gpi/IL-21 had significantly fewer nodules in the lung and lower lung weight [(285.8 ± 19.01) mg vs.(406.3 ± 27.12) mg],with lower levels of TGF-β2,ZEB1 and Ncadherin proteins but higher level of E-cadherin protein within the tumor tissue,as compared with the control mice.Meanwhile,the immunized mice had significantly increased CD8 + T cell killing activity [(42.62 ± 3.465) ％ vs.(22.29 ± 1.804) ％] and IFN-γexpression level [(55.200 ± 7.173) pg/ml vs.(6.435 ± 1.339) pg/ml] over the control mice.Conclusions The B16F10-ESAT-6-gpi/IL-21 vaccine can inhibit the metastasis of melanoma in the lung in vaccinated melanoma-bearing mice.This inhibitory effect is associated with CD8 + T cell immune response and a higher level of IFN-γ,which may influence on the mesenchymal-epithelial transition of tumor cells.