摘要
目的 研究前列腺癌细胞RNA转染树突状细胞(DCs)诱导的细胞毒T淋巴细胞(CTL)对前列腺癌细胞的特异性杀伤作用。方法联合应用粒细胞-巨噬细胞集落刺激因子(GM-CSF)、白细胞介素-4(IL-4)诱导培养小鼠骨髓单个核细胞向DCs分化,脂质体法转染RNA,流式细胞术测定转染后树突状细胞共刺激分子表达的变化,MTT法测定DCs对T淋巴细胞的刺激能力。ELISA法检测IFN-γ的分泌水平。LDH法测定DCs诱导的CTL对前列腺癌细胞的杀伤效果。结果用GM-CSF和IL-4诱导培养小鼠骨髓来源的单核细胞,体外诱导培养6 d后获得大量DCs;RNA转染后,DCs表面共刺激分子CD40、CD80、CD86和MHC-Ⅱ分子表达提高(P<0.05),刺激T淋巴细胞增殖能力明显提高(P<0.01);前列腺癌RNA转染的DCs诱导CTL对前列腺癌细胞和肝癌细胞的杀伤作用有明显差异(P<0.01)。结论前列腺癌细胞RNA转染DCs后诱导的CTL对前列腺癌有特异性杀伤作用,为进一步抗肿瘤疫苗的实验研究奠定了基础。
Objective To study RNA-DC vaccine inducing specific CTL against prostate cancer.Methods Mononuclear cells were induced into dendritic cells by cultured in GM-CSF and IL-4,RNA transfecting into DC by liposome.The surface molecules of DCs were analyzed by flow cytometry,IFN-γ levels in culture supernatant of DCs were measured by ELISA.The ability of DC to induce proliferation of autologous T lymphocytes was evaluated by MTT assay.The CTL antitumor activity in vitro induced by RNA-DC against prostate cancer cell was detected by LDH.Results After being cultured in GM-CSF and IL-4 for 6 days,A large number of morphologically typical dendritic cells were acquired.DCs highly expressed costimulatory molecules CD_（40）,CD_（80）,CD_（86） and MHC-Ⅱ after transfection（P0.05）,DC effectively stimulated proliferation of autologous T cells after transfection,the stimulating effect was significantly higher than those of untransfected DC.CTL induced by RNA modified DC had significantly higher cytotoxicity against RM-1 cells compared with induced by other DCs,and specifically kill RM-1 cell（P0.01）.Conclusion RNA-DC vaccine mediated by liposome could induce specific CTL against prostate cancer,and thus will lay a foundation for future research in producing the anti-tumor vaccine.
出处
《新疆医科大学学报》
CAS
2011年第6期577-581,共5页
Journal of Xinjiang Medical University
基金
新疆维吾尔自治区自然科学青年基金项目(2010211B22)
