南方医科大学学报2016，Vol.36Issue(10)：1423-1428,6.DOI:10.3969/j.issn.1673-4254.2016.10.22
RITA协同替莫唑胺抑制人脑胶质瘤U87细胞的生长
RITA combined with temozolomide inhibits the proliferation of human glioblastoma U87 cells
摘要
Abstract
Objective To observe the effect of RITA, a small molecule that targets p53, combined with temozolomide (TMZ) on proliferation, colony formation and apoptosis of human glioblastoma U87 cells and explore the underlying mechanism. Methods Cultured U87 cells were treated with RITA (1, 5, 10, 20μmol/L), TMZ, or RITA+TMZ (half dose) for 24, 48 or 72 h. MTS assay were used to detect the cell proliferation, and the cell proliferation rate and inhibitory rate were calculated. The effect of combined treatments was evaluated by the q value. The expressions of p53, p21 and other apoptosis-associated genes were detected by qRT-PCR and Western blotting;cell apoptosis was assayed using flow cytometry with Annexin V/PI double staining;colony formation of the cells was detected with crystal violet staining. Results MTS assay showed that RITA at the 4 doses more potently inhibited U87 cell viability than TMZ at 72 h (P=0.000) with inhibitory rates of 25.94%-41.38%and 3.84%-8.20%, respectively. RITA combined with TMZ caused a more significant inhibition of U87 cells (29.21%-52.11%) than RITA (P<0.01) and TMZ (P=0.000) alone. At the doses above 5μmol/L, the combined treatments with RITA+TMZ for 48 h resulted in q values exceeding 1.2 and showed an obvious synergistic effect of the drugs. Both RITA and TMZ, especially the latter, significantly increased the expressions of p53, p21, puma, and other apoptosis-associated genes to accelerate apoptosis and inhibit the growth and colony formation of U87 cells, and the effect was more obvious with a combined treatment. Conclusion RITA inhibits the growth of human glioblastoma cells and enhance their sensitivity to TMZ by up-regulating p53 expression, and when combined, RITA and TMZ show a synergistic effect to cause a stronger cell inhibition.关键词
人脑胶质瘤/RITA/替莫唑胺/肿瘤抑制Key words
human glioma/RITA/temozolomide/tumor inhibition引用本文复制引用
何小艳,冯小丽,宋鑫培,曾焕超,曹中栩,肖维威,张宝,吴清华..RITA协同替莫唑胺抑制人脑胶质瘤U87细胞的生长[J].南方医科大学学报,2016,36(10):1423-1428,6.基金项目
国家自然科学基金（81301911）；高等学校博士学科点专项科研基金（20124433120024） Supported by National Natural Science Foundation of China (81301911) （81301911）
