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目的:观察苦参碱(Matrine)对人胶质瘤细胞U251生长抑制作用及作用前后U251细胞对NK细胞杀伤敏感性的变化。方法:CCK-8法和台盼蓝染色法检测苦参碱对U251细胞生长抑制作用和细胞存活率;流式细胞仪法检测作用前后U251细胞周期变化及表面NKG2D配体(MICA/B、ULBP1-3)和HLA-Ⅰ类分子表达;乳酸脱氢酶释放法检测苦参碱作用前后U251细胞对NK细胞杀伤敏感性的变化。结果:苦参碱能抑制U251细胞生长,随着药物浓度增大和作用时间延长,抑制率逐渐升高、存活率逐渐下降;苦参碱作用后细胞发生G1/S期阻滞;细胞表面NKG2D各配体表达率明显升高,与作用前相比差异有统计学意义(P<0.05),HLA-Ⅰ类分子的表达率无明显变化(P>0.05);效靶比5:1、10:1、20:1时,作用前后U251细胞对NK细胞杀伤敏感性差异均有统计学意义(P<0.05)。结论:苦参碱能抑制U251细胞的生长,改变其周期,上调U251细胞表面NKG2D各配体表达率,增强其对NK细胞的杀伤敏感性。
OBJECTIVE: To observe the inhibitory effect of Matrine on the growth of human glioma U251 cells and the changes of U251 cell killing susceptibility to NK cells before and after treatment. METHODS: CCK-8 and trypan blue staining were used to detect the growth inhibitory effect and cell survival rate of matrine on U251 cells. Flow cytometry was used to detect the change of U251 cell cycle and surface NKG2D ligands (MICA/B, ULBP1) before and after treatment. -3) and HLA-I molecules expression; lactate dehydrogenase release assay to detect the effect of matrine on U251 cell killing susceptibility to NK cells. RESULTS: Matrine inhibited the growth of U251 cells. With the increase of drug concentration and prolonged action time, the inhibition rate gradually increased and the survival rate gradually decreased. After matrine exposure, G1/S phase arrest occurred; The expression rate of ligand was significantly increased compared with that before the effect (P<0.05). The expression rate of HLA-I molecules was not significantly changed (P>0.05); the ratio of effect to target was 5:1, 10: At 1 and 20:1, the sensitivity of U251 cells to NK cell killing before and after treatment was statistically significant (P<0.05). CONCLUSION: Matrine can inhibit the growth of U251 cells, change its cycle, up-regulate the expression rate of NKG2D ligands on U251 cells, and increase its sensitivity to NK cells.