乳腺癌细胞NK细胞活化性配体表达和CIK细胞杀伤敏感性研究

Research on Ligand Expression of NK Cell Activation in Breast Cancer Cells and Sensitivity of CIK Cells

目的研究乳腺癌细胞系MCF-7和MDA-MB-231细胞NK细胞活化性配体表达和细胞因子诱导的杀伤细胞(CIK)杀伤敏感性。方法采用流式细胞术检测MCF-7和MDA-MB-231细胞表面MICA、MICB、CD155和CD112的表达,PBMC在体外经γ-干扰素(IFN-γ),CD3抗体和IL-2诱导扩增为CIK细胞,乳酸脱氢酶释放法检测CIK细胞对MCF-7和MDA-MB-231细胞的杀伤活性。结果 MICA、MICB、CD155和CD112在MCF-7细胞表面的表达水平均明显高于MDA-MB-231细胞(P<0.01);培养15 d后,CIK细胞中CD3+CD56+细胞百分率显著增高(P<0.01);效靶比20:1时,CIK细胞对MDA-MB-231细胞的杀伤率高于其对MCF-7细胞的杀伤率,差异显著(P<0.01)。结论 CIK细胞对MCF-7和MDA-MB-231的杀伤活性不依赖于NK细胞活化性配体分子MICA、MICB、CD155和CD112的高表达;CIK过继免疫细胞治疗可能成为三阴乳腺癌的有效治疗手段。

Objective To study the ligand expressions of NK cell（ nature killer cell） activation in MCF-7 and MDA-MB-231 cells in breast carcinoma cell line and sensitivity of CIK（ cytokine-induced killer） cells. Methods The expressions of MICA,MICB,CD155 and CD112 on MCF-7 and MDA-MB-231 cells were detected by flow cytometry.CIK cells were obtained by the sequential ex vivo expansion of peripheral blood mononuclear cells（ PBMC） with interferon-gamma（ IFN-γ）,and anti-CD3 antibody and recombinant human interleukin（ IL）-2. The cytotoxicity of CIK cells against MCF-7 and MDA-MB-231 cells were detected by lactate dehydrogenase（ LDH） release assay. Results The expressions of MICA,MICB,CD155 and CD112 on MCF-7 cells were significantly higher than those on MDA-MB-231 cells（ P〈0. 01）; after 15 d of ex vivo expansion,the percentage of CD3 ＋ CD56 ＋ cell increased significantly（ P〈0. 01）;at E： T ratios of 20： 1,the cytotoxicity of CIK cells against MDA-MB-231 was significantly higher than that against MCF-7（ P〈0. 01）. Conclusion The cytotoxicity of CIK cells against MCF-7 and MDA-MB-231 cells is independent of overexpressions of MICA,MICB,CD155 and CD112. CIK adoptive immunity of cell therapy may be an effective approach for treatment of triple-negative breast cancer.