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目的探讨p53靶向小分子RITA对不同p53状态人脑胶质瘤细胞的生长抑制作用。方法基因测序确定实验所用U251和U87细胞中p53基因状态;MTS法检测0μmol/L、1μmol/L、4μmol/L、10μmol/L RITA作用于U251和U87细胞0 h、24 h、72 h、120 h、168 h的增殖情况;qRT-PCR法和Western blot法分别检测抑癌基因p53、p21的mRNA和蛋白质表达水平;计算抑制率和IC50。结果基因测序结果表明实验所用U251细胞表达突变型p53基因而U87表达野生型;RITA对2种细胞的增殖均有显著抑制作用,且对U251的抑制作用更强;5μmol/L RITA作用下,U251中p53、p21表达均显著增加,U87中p21表达增加,p53 mRNA表达变化不明显,但p53蛋白累积显著增加;RITA作用于U251、U87细胞24 h和120 h的IC50分别为123.870μmol/L、776.682μmol/L和2.817μmol/L、7.147μmol/L。结论RITA能有效促进人脑胶质瘤细胞U251和U87中p53和p21基因表达,发挥高效生长抑制作用,且含突变型p53的U251比含野生型p53的U87效果显著。
Objective To investigate the inhibitory effect of p53 targeted small molecule RITA on the growth of human glioma cells with different p53 status. METHODS: The gene status of p53 gene in U251 and U87 cells was determined by gene sequencing. MTS assay was performed on U251 and U87 cells at 0, 24, 72 and 120 h after RTA treatment with 0, 1, 4 and 10 μmol / h and 168 h respectively. The mRNA and protein expression levels of the tumor suppressor genes p53 and p21 were detected by qRT-PCR and Western blot respectively. The inhibition rates and IC50 were calculated. Results The gene sequencing results showed that U251 cells expressed mutant p53 gene and U87 expressed wild type. RITA significantly inhibited the proliferation of both cell lines and inhibited U251. Under the action of 5 μmol / L RITA, U251 The expression of p53 and p21 were significantly increased in U87, the expression of p21 was increased in U87, but the expression of p53 mRNA was not significantly changed, but the accumulation of p53 protein was significantly increased. The IC50 of RITA on U251 and U87 cells at 24 h and 120 h were 123.870 μmol / L, 776.682 micromol / L and 2.817 micromol / L, 7.147 micromol / L. Conclusions RITA can effectively promote the expression of p53 and p21 genes in human glioma cells U251 and U87 and exert a highly efficient growth inhibitory effect. U251 with mutant p53 has a significantly higher effect than U87 with wild-type p53.