CCND1表达沉默促进胶质母细胞瘤细胞株对替莫唑胺的敏感性

发布时间：2018-05-23 09:24

本文选题：细胞周期蛋白D基因 + 胶质母细胞瘤　；参考：《第二军医大学学报》2017年12期

【摘要】：目的利用已构建的CCND1表达沉默及过表达的人源性胶质母细胞瘤细胞株SHG-44,筛选能够与CCND1协同抑制胶质母细胞瘤细胞增殖的化学治疗药物。方法用蛋白质印迹法检测CCND1表达沉默及过表达的人源性胶质母细胞瘤细胞株SHG-44中多药耐药基因MDR1的表达产物P-糖蛋白(P-gp)及凋亡因子Bcl-2、Caspase-3的表达。使用卡莫司汀(BCNU)、洛莫司汀(CCNU)、替莫唑胺(TMZ)3种化学治疗药物分别处理CCND1过表达或表达沉默的SHG-44细胞,筛选能够与CCND1表达沉默协同抑制肿瘤细胞生长的化学治疗药物,并在人源性胶质母细胞瘤细胞株系U251中进一步验证。结果蛋白质印迹结果示CCND1表达沉默能够下调P-gp、Bcl-2的表达,上调Caspase-3的表达(P均0.01)。BCNU(0.05、0.25μg/mL)、CCNU(20、80μg/mL)处理CCND1过表达或表达沉默的SHG-44细胞的第2、3、4、5天时,细胞生长曲线的差异均无统计学意义;而在药物处理后细胞培养的第4、5天,CCND1表达沉默SHG-44细胞的生长受到TMZ(9.1μg/mL)的抑制作用较亲代SHG-44细胞强(P0.05)。U251细胞实验证实CCND1表达沉默促进TMZ的化学治疗敏感性。结论 CCND1表达沉默联合TMZ较两者单独作用能更有效地抑制人源性胶质母细胞瘤细胞株SHG-44的增殖,提示CCND1可能参与TMZ化学治疗耐药机制。
[Abstract]:Objective to screen the chemotherapeutic drugs which can inhibit the proliferation of glioblastoma cells in cooperation with CCND1 by using SHG-44, a human glioblastoma cell line with silencing and overexpression of CCND1. Methods the expression of multidrug resistance gene (MDR1) and apoptosis factor Bcl-2Caspase-3 in human glioblastoma cell line SHG-44 were detected by Western blot. Three chemotherapeutic agents, Carmosine, Lomoxetine, and temozolidazole, were used to treat SHG-44 cells with overexpression or silencing of CCND1, respectively, and to screen chemotherapeutic drugs which could inhibit the growth of tumor cells synergistically with CCND1 expression silencing. It was further verified in human glioblastoma cell line U251. Results Western blotting showed that CCND1 expression silencing could down-regulate the expression of P-gpnc-Bcl-2, and there was no significant difference in the growth curve of SHG-44 cells with CCND1 overexpression or silencing on the 2nd day after treatment of CCND1 overexpression or silencing SHG-44 cells (P < 0.01, P = 0.05, P < 0.05 or 0.25 渭 g / mL). On the 5th day after drug treatment, the growth of SHG-44 cells was inhibited by TMZ(9.1 渭 g / mL. the results showed that CCND1 expression silencing promoted the chemotherapeutic sensitivity of TMZ. Conclusion CCND1 expression silencing combined with TMZ can inhibit the proliferation of human glioblastoma cell line SHG-44 more effectively than both alone, suggesting that CCND1 may be involved in the mechanism of TMZ chemotherapeutic resistance.
【作者单位】：第二军医大学长征医院神经外科上海市神经外科研究所;火箭军总医院普外科;
【分类号】：R739.4

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