摘要
目的:研究微小RNA-422a(miR-422a)靶向调控人性别决定区Y框6(SOX6)对过氧化氢(H 2O 2)刺激下大鼠肾上腺嗜铬细胞瘤细胞株PC12(具有神经细胞特性)损伤的作用。方法:采用real-time PCR检测H 2O 2处理后PC12细胞中miR-422a表达的变化,在PC12细胞中转染miR-422a mimics,MTT法测定细胞活力,试剂盒检测乳酸脱氢酶(LDH)漏出率,流式细胞术测定细胞凋亡水平。靶基因预测软件预测SOX6可能是miR-422a的靶基因,萤光素酶报告基因实验鉴定靶向关系。在PC12细胞中共转染miR-422a mimics和SOX6过表达载体,检测过表达SOX6对miR-422a mimics调控H 2O 2条件下PC12细胞活力、LDH漏出率和凋亡的影响。结果:H 2O 2处理后PC12细胞中miR-422a表达水平降低(P<0.05)。H 2O 2处理后的PC12细胞的活力降低,LDH漏出率和细胞凋亡率升高(P<0.05)。miR-422a mimics能够提高H 2O 2条件下PC12细胞活力,降低LDH漏出率和凋亡率(P<0.05)。miR-422a靶向负调控SOX6表达。SOX6过表达载体可以逆转miR-422a对PC12细胞活力提高和LDH漏出率、细胞凋亡率降低的作用(P<0.05)。结论:miR-422a通过靶向抑制SOX6可降低H 2O 2对PC12细胞的损伤作用。
AIM:To investigate the effects of microRNA-422a(miR-422a)on the damage of rat adrenal gland pheochromocytoma PC12 cells induced by hydrogen peroxide(H 2O 2).METHODS:The expression of miR-422a in the PC12 cells treated with H 2O 2 was detected by real-time PCR.After miR-422a mimics were transfected into PC12 cells,the cell viability was measured by MTT assay,the lactate dehydrogenase(LDH)leakage rate was detected,and the apoptosis was analyzed by flow cytometry.Target gene prediction software was used to predict that sex-determining region Y box 6(SOX6)may be the target gene of miR-422a.Luciferase reporter assay was used to identify the targeting relationship.miR-422a mimics and SOX6 over-expression vector were co-transfected into the PC12 cells.The effects of SOX6 over-expression on the viability,LDH leakage rate and apoptosis of PC12 cells treated with H 2O 2 and transfected with miR-422a mimics were evaluated.RESULTS:The expression of miR-422a in the PC12 cells was decreased after treatment with H 2O 2(P<0.05).The viability of PC12 cells treated with H 2O 2 was decreased,and the LDH leakage rate and apoptotic rate were increased.Transfection with miR-422a mimics enhanced the viability of PC12 cells treated with H 2O 2,and the leakage rate of LDH and apoptotic rate of the PC12 cells were reduced.The expression of SOX6 was negatively regulated by miR-422a.SOX6 over-expression reversed the effects of miR-422a on PC12 cell viability,LDH leakage and apoptosis.CONCLUSION:miR-422a reduces the damage of PC12 cells induced by H 2O 2 via targeting SOX6.
作者
张广华
铁婷婷
巨虎
许常林
肖宗宇
苑乐
ZHANG Guang-hua;TIE Ting-ting;JU Hu;XU Chang-lin;XIAO Zong-yu;YUAN Le(Department of Neurosurgery,Affiliated Hospital of Qinghai University,Xining 810001,China;Department of Neurology,Affiliated Hospital of Qinghai University,Xining 810001,China;Teaching Management Department,Affiliated Hospital of Qinghai University,Xining 810001,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2019年第12期2235-2240,共6页
Chinese Journal of Pathophysiology
基金
青海大学附属医院中青年科技基金项目(No.ASRF-2015-ZD-01)
