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    抑制USP22表达对胶质瘤细胞顺铂耐药的影响

    Effect of inhibiting the expression of USP22 on cisplatin resistance in glioma cells

      摘要
    • 摘要:
      目的 探讨泛素特异性肽酶22（ubiquitin specific peptidase 22，USP22）对胶质瘤细胞顺铂耐药的作用。
      方法 采用顺铂剂量梯度爬升筛选方法诱导顺铂耐药胶质瘤细胞株U251/CP2，转染USP22小干扰RNA沉默胶质瘤顺铂耐药细胞株U251/CP2中USP22的表达，RT-PCR检测细胞USP22 mRNA的表达水平，流式细胞术检测转染USP22-shRNA对细胞凋亡的影响，CCK-8法检测细胞的活力及耐药指数，Western blotting检测USP22的蛋白水平。
      结果 成功构建了顺铂耐药胶质瘤细胞株U251/CP2、USP22基因沉默细胞株U251-shUPS22和U251/CP2-shUSP22。抑制USP22的表达可以明显提高U251/CP2-shUSP22细胞的凋亡率（P < 0.05），顺铂上调USP22蛋白在U251和U251/CP2细胞中的表达（P < 0.01）。U251-shUPS22、U251/CP2和U251/CP2-shUSP22各组细胞的顺铂半数抑制浓度分别为（0.56±0.07）μg/mL、（73.73±2.74）μg/mL和（51.25±1.09）μg/mL，与对照U251组（1.23±0.13）μg/mL比较差异均有统计学意义（P < 0.01）。
      结论 抑制USP22表达可增强胶质瘤细胞对顺铂的敏感性，且可在一定程度上逆转耐药细胞株的顺铂耐药，提示USP22是胶质瘤顺铂耐药的潜在分子机制之一。

       

      Abstract:
      Objective To investigate the effects of inhibiting the expression of ubiquitin specific peptidase 22(USP22) on cisplatin resistance in glioma cells.
      Methods The cisplatin-resistant glioma cell line(U251/CP2) was induced by gradual elevation of cisplatin concentration.The expression of USP22 in glioma cell line U251/CP2 was silenced using the transfection of USP22 small interfering RNA.The expression level of USP22 mRNA, cell apoptosis, viability and drug resistance index and protein level of USP22 were detected using RT-PCR, flow cytometry, CCK-8 and Western blotting, respectively.
      Results The cisplatin-resistant glioma cell line of U251/CP2 and USP22 gene silencing cell lines of U251-shUPS22 and U251/CP2-shUSP22 were successfully constrcucted.The inhibition of USP22 expression could significantly increase the apoptosis rate of U251/CP2-shUSP22 cells(P < 0.01), and cisplatin could up-regulate the expression of USP22 protein in U251 and U251/CP2 cells(P < 0.01).The IC50 values to cisplatin of U251-shUPS22, U251/CP2, U251/CP2-shUSP22 and U251 cells were (0.56±0.07)μg/mL, (73.73±2.74)μg/mL, (51.25±1.09)μg/mL and (1.23±0.13)μg/mL, respectively.
      Conclusions The inhibition of USP22 expression can enhance the chemosensitivity to cisplatin in glioma cells, and reverse the resistance of cisplatin in resistant cell line.It is suggested that USP22 may be one of the potential molecular mechanisms of cisplatin resistance in glioma.

       

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