摘要
目的探究长链非编码(lncRNA)RPA3-AS1影响人心肌细胞凋亡的作用机制。方法采用实时定量聚合酶链反应(qPCR)技术检测43例急性心肌缺血患者和43例体检健康者全血样本中RPA3-AS1的表达。分别转染载有RPA3-AS1序列的质粒和阴性对照质粒至人心肌细胞系AC16,设为实验组和对照组。采用qPCR确定转染效率。细胞增殖实验(MTT法)和流式细胞术分别检测两组AC16细胞的活力和凋亡情况。生物信息学方法预测RPA3-AS1的作用机制。qPCR和Western blot法分别检测RPA3-AS1靶基因的表达。结果与体检健康者相比,急性心肌缺血患者全血中RPA3-AS1的表达明显降低,差异有统计学意义(P<0.01)。与对照组相比,实验组AC16细胞中RPA3-AS1的表达显著升高,差异有统计学意义(P<0.01),细胞活力显著增加,差异有统计学意义(P<0.05),细胞凋亡率显著降低,差异有统计学意义(P<0.05)。RPA3-AS1的靶标可能是微小RNA-203a-3p(miR-203a-3p),miR-203a-3p的靶基因可能是Bcl-2相关抗凋亡蛋白3(BAG3)。与对照组相比,实验组AC16细胞中miR-203a-3p的表达显著降低,差异有统计学意义(P<0.01),BAG3基因的表达显著升高,差异有统计学意义(P<0.01)。结论RPA3-AS1可能通过影响miR-203a-3p/BAG3分子轴,促进人心肌细胞的活力并抑制其凋亡,从而保护人心肌细胞。
Objective To explore the mechanism of long non coding RNA(lncrna)rpa3-as1 on human cardiomyocyte apoptosis.Methods Real-time quantitative polymerase chain reaction(qPCR)was used to detect the expression of RPA3-AS1 in the whole blood samples of 43 patients with acute myocardial ischemia and 43 healthy individuals.The plasmid carrying the RPA3-AS1 sequence and the negative control plasmid were respectively transfected into the human cardiomyocyte cell line AC16,named the experimental group and the control group,and the transfection efficiency was determined by qPCR.Cell proliferation test(MTT method)and flow cytometry were used to detect the viability and apoptosis of two groups of human cardiomyocytes.Bioinformatics methods predict the mechanism of action of RPA3-AS1.QPCR and Western blot methods were used to detect the expression of RPA3-AS1 target genes.Results Compared with healthy people who underwent physical examination,the expression of RPA3-AS1 in the whole blood of patients with acute myocardial ischemia was significantly reduced,and the difference was statistically significant(P<0.01).Compared with the control group,the expression of RPA3-AS1 in the AC16 human cardiomyocyte cell line of the experimental group was significantly increased,and the difference was statistically significant(P<0.01),cell viability was significantly increased,and the difference was statistically significant(P<0.05),and the apoptosis rate was significantly reduced,and the difference was statistically significant(P<0.05).The target of RPA3-AS1 may be miR-203a-3p,and the target gene of miR-203a-3p may be Bcl-2 associated athanogene 3(BAG3).Compared with the control group,the expression of miR-203a-3p in the AC16 human cardiomyocyte cell line of the experimental group was significantly reduced,and the difference was statistically significant(P<0.01),and the expression of BAG3 gene was significantly increased,and the difference was statistically significant(P<0.01).Conclusion RPA3-AS1 may regulate the miR-203a-3p/BAG3 molecular axis
作者
黄锐
王剑
谭慧
周利平
杨化冰
刘娟
HUANG Rui;WANG Jian;TAN Hui;ZHOU Liping;YANG Huabing;LIU Juan(Department of Cardiovascular Medicine,Lichuan People′s Hospital,Lichuan,Hubei 445400,China;Department of Pharmacy of Western Medicine,Enshi Central Hospital,Enshi,Hubei 445000,China)
出处
《国际检验医学杂志》
CAS
2021年第14期1730-1734,共5页
International Journal of Laboratory Medicine
基金
2019年湖北省卫生计生委中医药科研面上项目(ZY2019Z001)。
