摘要
目的通过检测结肠癌细胞株HCT-8及长春新碱(VCR)耐药细胞株HCT-8/VCR中微小核糖核酸(miRNA)的表达差异,分析miRNA与结肠癌细胞耐药的关系。方法采用TRIzol方法提取结肠癌细胞株HCT-8和耐药细胞株HCT-8/VCR的总RNA,用紫外分光光度计法检测提取的RNA纯度并计算其浓度,琼脂糖凝胶电泳确定其完整性,反转录后采用实时荧光定量聚合酶链反应法检测HCT-8和HCT-8/VCR细胞中hsa-miRNA-126-3p、hsa-miRNA-7974及hsa-miRNA-6847-5p的相对含量。结果人结肠癌敏感细胞株HCT-8、结肠癌耐药细胞株HCT-8/VCR在260 nm处总RNA分别为763、245 mg·L^(-1),A260/A280比值分别为1.96、2.10;HCT-8/VCR细胞中的hsa-miRNA-126-3p、hsa-miRNA-7974和hsa-miRNA-6847-5p的相对含量均高于HCT-8细胞(P<0.01,P<0.001)。结论 miRNA的异常表达与结肠癌细胞VCR耐药存在一定关系。
Objective To analyze the relationship between microRNA（ miRNA） and the drug resistance of colon cancer cell by detecting the expression of miRNA in human colon cancer cell line HCT-8 and vincristine（ VCR）-resistant HCT-8（ HCT-8 /VCR）. Methods Total RNA in HCT-8 and HCT-8 /VCR cells was extracted by TRIzol,the purity of RNA was measured by ultraviolet spectrophotometer,and the integrality of RNA was identified by agarose gel eletrophoresis,then reverse transcription was performed. The relative expression levels of hsa-miRNA-126-3p,hsa-miRNA7974 and hsa-miRNA-6847-5p in HCT-8 and HCT-8/VCR were detected by quantitative real-time polymerase chain reaction. Results The total RNA of HCT-8 and HCT-8 / VCR at 260 nm was 763 mg·L^（-1） and 245 mg·L^（-1） respectively,the A260/ A280 of RNA was 1. 96 and 2. 10 respectively. The relative expression of hsa-miRNA-126-3p,hsa-miRNA7974 and hsa-miRNA-6847-5p in HCT-8 / VCR were significantly higher than those in HCT-8（ P 〈 0. 01,P 〈 0. 001）. Conclusion The abnormal expression of miRNA was associated with VCR resistance of colon cancer cells.
出处
《新乡医学院学报》
CAS
2017年第3期170-172,共3页
Journal of Xinxiang Medical University
