重组人精氨酸酶对人脑胶质瘤细胞U87的杀伤作用研究

发布时间：2018-03-25 07:19

本文选题：重组人精氨酸酶　切入点：人脑胶质瘤　出处：《中国临床药理学杂志》2017年02期

【摘要】：目的研究重组人精氨酸酶对人脑胶质瘤细胞U87的杀伤作用。方法培养人脑胶质瘤细胞U87,将对数生长期人脑胶质瘤细胞U87随机分为12组:对照组仅加等量的培养液,其余11组实验组以倍比稀释的方式分别给予质量浓度为0.2×10~(-3),0.5×10~(-3),0.1×10~(-2),0.2×10~(-2),0.4×10~(-2),0.8×10~(-2),1.6×10~(-2),3.2×10~(-2),6.3×10~(-2),1.25×10-1,0.25 U·mL~(-1)的重组人精氨酸酶,用甲基噻唑蓝四氮唑盐(MTT)法检测各组人脑胶质瘤细胞U87细胞活力的变化;倒置显微镜观察U87细胞经重组人精氨酸酶作用后的形态学变化;Western blotting法检测精氨酸琥珀酸合成酶(ASS)的表达。重组人精氨酸酶作用于人脑胶质瘤细胞U87时,分成补充L-精氨酸组和不补充L-精氨酸组,用MTT法检测人脑胶质瘤细胞U87细胞活力的变化。结果不同质量浓度的重组人精氨酸酶作用U87细胞72 h后,U87细胞的活力均降低且呈明显的剂量依赖性,0.2×10~(-2),0.8×10~(-2),1.6×10~(-2),0.25 U·mL~(-1)实验组的细胞活力分别为75.12%,52.28%,44.09%,38.14%,差异有统计学意义(P0.05)。通过倒置显微镜观察,重组人精氨酸酶作用U87细胞72 h后,0.2×10~(-2),1.6×10~(-2),1.25×10-1U·mL~(-1)实验组的细胞密度随着药物浓度升高而显著降低,镜下可见U87细胞形态发生明显的变形,皱缩,坏死。Western blotting实验结果表明,U87细胞的ASS蛋白表达缺陷。当重组人精氨酸酶作用于U87细胞同时,补充部分L-精氨酸时的细胞活力为60.53%,未补充L-精氨酸组的细胞活力为47.48%,差异有统计学意义(P0.01)。结论由于人脑胶质瘤细胞U87细胞合成精氨酸的关键酶ASS蛋白表达缺陷,重组人精氨酸酶通过耗竭外源性精氨酸,对人脑胶质瘤细胞U87产生显著的杀伤效应。
[Abstract]:Objective to study the killing effect of recombinant human argininase on human glioma cell U87. Methods Human glioma cell U87 in logarithmic phase was randomly divided into 12 groups: the control group was treated with the same amount of culture medium. The other 11 experimental groups were given the recombinant human argininase with a mass concentration of 0.2 脳 10 ~ (-10) ~ (-3) and 0.5 脳 10 ~ (-1) U ~ (-1). The activity of U87 cells in each group was detected by MTT method. The activity of U87 cells in each group was detected by MTT- (0. 8 脳 10 ~ (-2)) 脳 10 ~ (-10) ~ (2) ~ (3) 脳 10 ~ (-10) ~ (2) ~ (2). The morphological changes of U87 cells treated with recombinant human argininase were observed by inverted microscope. The expression of arginine succinate synthase (ASS) in U87 cells was detected by Western blotting. Divided into L-arginine supplementation group and non-L-arginine supplementation group, The activity of U87 cells was detected by MTT assay. Results the activity of U87 cells decreased in a dose-dependent manner after 72 h treatment with different concentrations of recombinant human argininase. The activity of U87 cells decreased in a dose-dependent manner. The cell viability was 75.12 and 52.28 and 44.09 and 38.14, respectively. The difference was statistically significant (P 0.05). After 72 hours of treatment with recombinant human argininase, the cell density of U87 cells decreased significantly with the increase of drug concentration, and U87 cells were morphologically deformed and shrinked under microscope. The results of Western blotting assay showed that the expression of ASS protein was deficient in U87 cells. The cell viability of L- arginine supplementation group was 60.53, and that of non-L-arginine group was 47.48. The difference was statistically significant (P 0.01). Conclusion the key enzyme ASS protein expression of arginine synthesis in human glioma cell U87 is defective. Recombinant human argininase exerts significant killing effect on human glioma cell line U87 by exhausting exogenous arginine.
【作者单位】：南方医科大学南方医院药物临床试验中心;
【基金】：国家自然科学基金资助项目(U1401226)
【分类号】：R739.41

【相似文献】