miR-520a-5p通过调控有氧糖酵解诱导前列腺癌细胞多西他赛耐药  

miR-520a-5p induces docetaxel resistance in prostate cancer cells by regulating aerobic glycolysis

作　　者：谢剑云[1] 陈伟东[1] Xie Jianyun;Chen Weidong(Department of Urology,The People's Hospital Affiliated to Fujian University of Traditional Chinese Medicine)

机构地区：[1]福建中医药大学附属人民医院泌尿外科,福州350004

出　　处：《重庆医科大学学报》2023年第8期909-915,共7页Journal of Chongqing Medical University

摘　　要：目的:探索mi R-520a-5p对前列腺癌增殖能力和多西他赛耐药性的影响及其作用机制。方法:使用RT-q PCR检测PC-3/DU145前列腺癌细胞及RWPE-1前列腺正常上皮细胞中mi R-520a-5p表达水平;使用免疫印迹检测mi R-520a-5p是否调控糖酵解关键酶己糖激酶2(hexokinase 2,HK2)和磷酸果糖激酶(phosphofructokinase,PFKM)表达水平;使用CCK-8实验检测mi R-520a-5p对前列腺癌细胞增殖能力和多西他赛耐药性的影响;使用ELISA评估培养上清中葡萄糖消耗水平和乳酸生成水平。依据转染物的不同,将所有细胞分成4组:minic组、minic NC组、inhibitor组、inhibitor NC组。结果:mi R-520a-5p在前列腺癌细胞中表达下调(P<0.001)。与minic NC组比较,minic组可显著降低PC-3和DU145前列腺癌细胞的增殖能力(P<0.001)、多西他赛半抑制浓度(IC_(50))、葡萄糖消耗水平和乳酸生成水平(P<0.001);与inhibitor NC组比较,inhibitor组可明显增强PC-3和DU145前列腺癌细胞的增殖能力(P<0.001)、葡萄糖消耗水平和乳酸生成水平(P<0.001)。Starbase数据库提示mi R-520a-5p与HK2和PFKM存在结合位点;免疫印迹结果显示,mi R-520a-5p对HK2和PFKM的表达存在负调控关系。使用2-DG抑制前列腺癌细胞糖酵解可恢复mi R-520a-5p对前列腺癌增殖能力的影响。结论:前列腺癌中mi R-520a-5p可能通过上调糖酵解关键酶HK2和PFKM表达水平增强前列腺癌细胞有氧糖酵解诱导多西他赛化疗耐药。Objective To explore the effects of miR-520a-5p on the proliferation and docetaxel resistance of prostate cancer cells and the potential mechanisms.Methods The expression levels of miR-520a-5p in prostate cancer cells(PC-3/DU145)and normal prostate epithelial cells(RWPE-1)were measured by RT-qPCR.Western blot was used to determine whether miR-520a-5p regulated the expression of the key glycolytic enzymes hexokinase 2(HK2)and phosphofructokinase(PFKM).The effects of miR-520a-5p on the proliferation and docetaxel resistance of prostate cancer cells were determined using cell counting kit-8.Glucose consumption and lactic acid production in the culture supernatant were assessed by enzyme-linked immunosorbent assay.According to the transfection,all cells were divided into 4 groups:minic group,minic NC group,inhibitor group and inhibitor NC group.Results miR-520a-5p was significantly down-regulated in prostate cancer cells(P<0.001).Compared with the mimic NC group,the mimic group showed significant decreases in the proliferative ability(P<0.001),half-maximal inhibitory concentration(IC50)of docetaxel,glucose consumption level,and lactic acid production level(P<0.001)of PC-3 and DU145 prostate cancer cells.Compared with the inhibitor NC group,the inhibitor group showed significantly increased proliferative ability(P<0.001)and glucose consumption and lactic acid production levels(P<0.001)of PC-3 and DU145 prostate cancer cells.Starbase database analysis indicated that there were binding sites between miR-520a-5p and HK2 and PFKM.Western blot results showed that miR-520a-5p negatively regulated the expression of HK2 and PFKM.Inhibiting the glycolysis of prostate cancer cells by 2-DG could recover the effect of miR-520a-5p on the proliferative ability of prostate cancer.Conclusion The low expression of miR-520a-5p in prostate cancer may up-regulate the key glycolytic enzymes HK2 and PFKM to enhance aerobic glycolysis,thereby inducing resistance to docetaxel in prostate cancer cells.

关 键 词：前列腺癌 有氧糖酵解 多西他赛 化疗耐药 

分 类 号：R737.25[医药卫生—肿瘤]