miR-499a-5p通过靶向Pten减轻嗜铬细胞瘤细胞PC12低氧损伤  

miR-499a-5p alleviates hypoxic damage of pheochromocytoma cells through targeting at Pten

作　　者：张丽阳 孙军[1] 陈迪 ZHANG Li-yang;SUN Jun;CHEN Di(Department of Neurology,Nanyang Central Hospital,Nanyang 473000,China)

机构地区：[1]南阳市中心医院神经内科,河南南阳473000

出　　处：《基础医学与临床》2022年第12期1873-1878,共6页Basic and Clinical Medicine

基　　金：河南省科学技术厅基金(192102310349)。

摘　　要：目的研究miR-499a-5p对化学性低氧诱导的嗜铬细胞瘤细胞(PC12)损伤的影响。方法将PC12细胞分为对照组、低氧组(CoCl_(2)处理24 h)、低氧+miR-NC组和miR-mimic转染组(分别转染miR-NC和miR-499a-5p mimic后,用CoCl_(2)处理24 h)。用RT-qPCR检测miR-499a-5p表达;用MTT法检测细胞活性;用试剂盒检测乳酸脱氢酶(LDH)活性;用流式细胞测量术检测各组细胞凋亡;用Western blot检测磷酸酶与张力蛋白同源物(PTEN)蛋白的表达。用荧光素酶报告基因检测Pten是否为miR-499a-5p的直接作用靶标。结果与对照组比较,低氧组中miR-499a-5p表达和细胞活性降低(P<0.01或P<0.001),LDH活性和细胞凋亡增加(P<0.01或P<0.001);过表达miR-499a-5p可提高低氧条件下PC12细胞的存活率并抑制其凋亡(P<0.05或P<0.01)。Pten是miR-499a-5p的直接作用靶标。结论miR-499a-5p通过靶向Pten减轻PC12细胞低氧损伤。Objective To study the effect of miR-499a-5p on injury induced by chemical hypoxia of pheochromocytoma cell(PC12).Methods PC12 cells were divided into control group,hypoxia group(treated with CoCl2 for 24 h),hypoxia+miR-NC or miR-mimic groups.Cell viability was detected by MTT assay;the expression of miR-499a-5p was detected by RT-qPCR;lactate dehydrogenase(LDH)activity was detected by kit;apoptosis was detected by flow cytometry;the expression of phosphatase and tensin homolog(PTEN)protein was detected by Western blot.The luciferase reporter gene was used to detect whether Pten was a direct target of miR-499a-5p.Results Compared with control group,the cell viability and miR-499a-5p expression decreased(P<0.01 or P<0.001),LDH activity and apoptosis increased(P<0.01 or P<0.001)in hypoxia group.Over-expression of miR-499a-5p enhanced survival rate and inhibited apoptosis of PC12 cells under hypoxia(P<0.05 or P<0.01).Pten was a direct target of miR-499a-5p.Conclusions miR-499a-5p alleviates hypoxic injury of PC12 cells through targeting at Pten.

关 键 词：miR-499a-5p 磷酸酶与张力蛋白同源物 PC12细胞 低氧损伤 

分 类 号：R966[医药卫生—药理学]