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引用本文:周红建,李军,夏建国.顺铂诱导脑胶质瘤U87细胞的PD-L1表达及耐药机制的研究[J].中国现代应用药学,2022,39(9):1155-1161.
ZHOU Hongjian,LI Jun,XIA Jianguo.Study on Expression of PD-L1 and Resistance Mechanism on Glioma U87 Cells Induced by Cisplatin[J].Chin J Mod Appl Pharm(中国现代应用药学),2022,39(9):1155-1161.
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顺铂诱导脑胶质瘤U87细胞的PD-L1表达及耐药机制的研究
周红建, 李军, 夏建国
杭州市萧山区中医院神经外科, 杭州 311201
摘要:
目的 研究PD-L1的表达对U87细胞的顺铂(cisplatin,CDDP)耐药性的影响。方法 建立U87对CDDP耐药细胞株(U87/CDDP),用不同浓度的CDDP处理24 h后,CCK-8检测U87/CDDP细胞活力以验证耐药性。成功建立U87/CDDP细胞株后转染PD-L1 shRNA,用CCK-8、BrdU染色、流式细胞仪检测U87/CDDP细胞的增殖和凋亡。并分别用qRT-PCR检测PD-L1、Bax、caspase-3、Survivin mRNA的表达,Western blotting检测PD-L1、Bax、caspase-3、cleaved-caspase-3和P53蛋白的表达。结果CCK-8结果表明,U87/CDDP细胞成功建立。用相同浓度的CDDP处理24 h后,转染PD-L1 shRNA的U87/CDDP细胞的细胞增殖率比未转染的U87/CDDP细胞低,细胞凋亡率高。同时,与对照组细胞相比,在转染PD-L1 shRNA并给予CDDP处理的U87/CDDP细胞中,Bax、caspase-3 mRNA和Bax、cleaved-caspase-3和P53蛋白的表达显著增加,PD-L1、Survivin mRNA和PD-L1蛋白的表达显著降低。结论 通过抑制U87细胞中PD-L1的表达可以逆转脑胶质瘤U87细胞对CDDP的耐药性。
关键词:  脑胶质瘤  U87细胞  顺铂  PD-L1  耐药性
DOI:10.13748/j.cnki.issn1007-7693.2022.09.004
分类号:R965.2
基金项目:萧山区重大科技攻关项目(2017309)
Study on Expression of PD-L1 and Resistance Mechanism on Glioma U87 Cells Induced by Cisplatin
ZHOU Hongjian, LI Jun, XIA Jianguo
Department of Neurosurgery, Hangzhou Xiaoshan District Hospital of Traditional Chinese Medicine, Hangzhou 311201, China
Abstract:
OBJECTIVE To investigate the effect of PD-L1 expression on the cisplatin(CDDP) resistance of U87 cells. METHODS U87 CDDP drug-resistant cell line(U87/CDDP) was established and the cell viability was estimated by CCK-8 to verify drug resistance after treated with different concentration of CDDP for 24 h. Then the U87/CDDP cells were transferred PD-L1 shRNA, and CCK-8, BrdU staining, flow cytometry was performed to detected the proliferation and apoptosis. Meanwhile, the mRNA expression of PD-L1, Bax, caspase-3, Survivin was detected by qRT-PCR, while the protein expression of PD-L1, Bax, caspase-3, cleaved-caspase-3, P53 was detected by Western blotting. RESULTS The result of CCK-8 showed that U87/CDDP was established successfully. After treated with same concentration of CDDP 24 h, the proliferation of PD-L1 shRNA-U87/CDDP cells was decreased and the apoptosis was increased compared with non transfected U87/CDDP cells. Meanwhile, compared to the control group, the mRNA expression of Bax, caspase-3 and the protein expression of Bax, cleaved-caspase-3 and P53 was significantly increased, while the mRNA expression of PD-L1, Survivin and the protein expression of PD-L1 was significantly decreased in PD-L1 shRNA transfected and CDDP treated U87/CDDP cells. CONCLUSION The tolerance of CDDP on U87 cells can reverse by down-regulating the expression of PD-L1.
Key words:  glioma  U87 cells  cisplatin  PD-L1  tolerance
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