Cbl-b-SiRNA转染淋巴细胞免疫杀伤MFC胃癌细胞的实验研究

Experimental study on immune function against MFC gastric cancer cells of lymphocyte transfected with Cbl-b-SiRNA in vitro

目的探讨cbl-b(Casitas B cell lymphoma-b)特异性小干扰RNA转染小鼠淋巴细胞后,能否增强其对MFC胃癌细胞的主动免疫杀伤作用。方法设计筛选cbl-b-SiRNA,小鼠脾脏淋巴细胞体外培养扩增,以脂质体为载体将cbl-b-SiRNA导入淋巴细胞,Real time PCR验证对cbl-b mRNA的表达抑制效率,用CCK-8试剂盒检测转染后的淋巴细胞对MFC胃癌细胞的杀伤作用。结果 cbl-b-SiRNA转染的淋巴细胞较未转染淋巴细胞,对MFC胃癌细胞的杀伤作用明显增强,高达(74.09±1.77)%。其中混合培养48 h的淋巴细胞杀伤率较72h增强。结论通过脂质体法可高效率将cbl-b-SiRNA转入淋巴细胞,且转染后的淋巴细胞对MFC胃癌细胞的主动免疫杀伤作用明显增强。

Objective To investigate whether lymphocytes transfected with cbl-b（Casitas B cell lymphoma-b）specific small interfering RNA are capable of improving the active immunization against MFC gastric cancer cells in vitro.Methods Cbl-b-siRNAs were designed and screened,spleen lymphocytes were cultured in vitro.The small interfering RNA（siRNA）targeting cbl-b was transfected into lymphocytes by lipofectamine TM2000.cbl-b mRNA expression was examined by real-time PCR.The cytotoxicity of lymphocytes against MFC gastric cancer cells were examined by CCK-8 assay kit in vitro.Results The lymphocytes,which were transfected with cbl-b-SiRNA,could significantly enhance the killing effect on the the MFC gastric cancer cells,up to（74.09±1.77）%.The lymphocytes,which mixed culture 48 h with gastric cancer cells has increased killing rate than that 72 h.Conclusion Cbl-b-siRNA can be transfered into spleen lymphocytes with Lipidosome efficiently,and active immunization of lymphocytes after transfection against MFC gastric cancer cells was significantly enhanced.