山东医药2017，Vol.57Issue(23)：8-11,4.DOI:10.3969/j.issn.1002-266X.2017.23.003
转染CDK2-shRNA对黑色素瘤B16-F1细胞达巴卡嗪敏感性的影响及其机制
Effect of transfection of CDK2-shRNA on chemotherapy sensitivity of melanoma B16-F1 cells to dacarbazine
摘要
Abstract
Objective To explore the effects of cyclin-dependent protein kinase 2-short hairpin RNA (CDK2-shRNA) on the chemotherapy sensitivity of B16-F1 cells to dacarbazine (DTIC) and its mechanism.Methods B16-F1 cells were divided into the observation group, empty plasmid group, and control group.The recombinant lentiviral vector pUL-CDK2-shRNA was transfected into the observation group, the recombinant plasmid pUL-NC-shRNA was transfected into the empty plasmid group, and the control group was not transfected with any plasmid.After 12 hours, every group was treated with 250 μmol/L dacarbazine for 72 h.MTT assay was used to detect the absorbance values in the three groups.Colony forming assay was used to detect colony forming rate (CFR).AnnexinV-FITC/PI double staining method was used to detect the apoptosis.Western blotting was used to detect the protein expression levels of Bcl-2, Bax, and Caspase-3 in each group, then we calculated Bax/Bcl-2 ratio.Results The absorbance values in the observation group, the empty plasmid group, and the control group were 0.221±0.019, 0.684±0.049, and 0.699±0.051, respectively.The CFR in the above three groups were 33.00%±1.80%, 67.50%±7.26%, and 69.83%±5.25% respectively.The apoptosis rates in the above three groups were 43.6%±4.1%, 17.6%±3.8%, and 15.1%±4.0%.The expression ratios of Bax/Bcl-2 in the above three groups were 2.56±0.28, 0.84±0.04, and 0.86±0.11, respectively.The relative expression of Caspase-3 was 0.92±0.07, 0.69±0.08, and 0.57±0.05, respectively.Significant differences were found in the absorbance value, CFR, apoptosis rate, and expression of Bax/Bcl-2 and Caspase-3 between the observation group and the empty plasmid group, the control group (all P<0.05).Conclusion The transfection of CDK2-shRNA can improve the chemotherapy sensitivity of melanoma B16-F1 cells to DTIC, and the mechanism may be that B16-F1 melanoma cells transfected with CDK2-shRNA in advance can increase the expression levels of Bax/Bcl-2 and Caspase-3, and then promote the apoptosis of melanoma B16-F1 cells.关键词
黑色素瘤/周期素依赖性蛋白激酶2/短发夹RNA/达卡巴嗪/药物疗法/耐药性/细胞凋亡Key words
melanoma/cyclin-dependent kinase 2/short hairpin RNA/dacarbazine/drug therapy/drug resistance/apoptosis分类
医药卫生引用本文复制引用
晋佳路,朱仁书,谢育媛,刘红春..转染CDK2-shRNA对黑色素瘤B16-F1细胞达巴卡嗪敏感性的影响及其机制[J].山东医药,2017,57(23):8-11,4.基金项目
河南省科技计划项目(122300410193) （122300410193）
河南省高等学校青年骨干教师资助计划项目(2011GGJS-262). （2011GGJS-262）
