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-L1shRNA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。
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-L1shRNA-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.
作者
周红建
李军
夏建国
ZHOU Hongjian;LI Jun;XIA Jianguo(Department of Neurosurgery,Hangzhou Xiaoshan District Hospital of Traditional Chinese Medicine,Hangzhou 311201,China)
出处
《中国现代应用药学》
CAS
CSCD
北大核心
2022年第9期1155-1161,共7页
Chinese Journal of Modern Applied Pharmacy
基金
萧山区重大科技攻关项目(2017309)。
