论文部分内容阅读
目的 研究导入NKG5SV基因对NK细胞肝癌杀伤作用的影响及其机理。方法 将NKG5SV和对照基因LacZ基因分别导入肝癌病人的NK细胞 ,K5 6 2细胞杀伤实验检测NK活性 ,MTT法检测不同NK细胞对肝癌细胞株和正常肝细胞株的杀伤 ,并进行形态学观察。DAPI荧光染色法和小片段DNA定量法检测和定量细胞凋亡。结果 肝癌病人NK细胞、NK LacZ细胞、NK NKG5SV细胞 ,健康人NK细胞的NK活性 (% )分别为 5 96± 0 38、6 0 3± 0 42 (P >0 0 5 )、2 7 6 7± 0 18(P <0 0 5 )、30 33± 0 83(P <0 0 5 ) 作用 72h后肝癌病人NK细胞、NK LacZ细胞、NK NKG5SV细胞对SMMC 772 1肝癌细胞株的杀伤活性 (% )分别为 45± 5 ,46± 9(P >0 0 5 )、6 9± 4(P <0 0 1)。SMMC 772 1细胞的凋亡细胞数 (个 /高倍视野 )和凋亡DNA量 (μg/板 )分别为 1 5± 0 3、1 4± 0 5 (P >0 0 5 )、1 9± 0 6 (P >0 0 5 ) ,70± 10、72± 9(P >0 0 5 )、76± 10 (P >0 0 5 )。结论 NKG5SV基因导入肝癌病人的NK细胞通过增加其细胞溶解作用而提高其肿瘤杀伤能力。
Objective To study the effect of the introduction of NKG5SV gene on the killing effect of NK cell carcinoma and its mechanism. Methods The NKG5SV and control gene LacZ gene were introduced into NK cells of HCC patients respectively. The K562 cell killing assay was used to detect NK activity. MTT assay was used to detect the killing of different NK cells to hepatoma cell lines and normal liver cell lines. Morphological observation was performed. DAPI fluorescent staining and small fragment DNA quantification were used to detect and quantify apoptosis. Results The NK activity (%) of NK cells, NK LacZ cells, NK NKG5SV cells, and healthy human NK cells were 5 96±0 38, 60 3±0 42 (P >0 0 5) and 2 7 6 7 respectively. The killing activity of NK cells, NK LacZ cells and NK NKG5SV cells against SMMC 772 1 hepatocellular carcinoma cell lines after 72 h exposure to ± 0 18 (P <0 0 5) and 30 33 ± 0 83 (P <0 05) 45 ± 5, 46 ± 9 (P > 0 0 5), and 6 9 ± 4 (P <0 0 1), respectively. The number of apoptotic cells (cells per high power field) and the amount of apoptotic DNA (μg/plate) of SMMC 772 1 cells were 1 5± 0 3, 1 4± 0 5 (P > 0 0 5 ), and 1 9± 0, respectively. 6 (P > 0 0 5), 70 ± 10, 72 ± 9 (P > 0 05), 76 ± 10 (P > 0 0 5). Conclusion The NKG5SV gene was introduced into NK cells of patients with liver cancer to increase its cytotoxicity by increasing its cytolysis.