H2O2诱导H9c2细胞凋亡中circNFIX表达及作用(1)
[摘要]目的 探討环状RNA NFIX(circNFIX)在H9c2细胞凋亡中的表达趋势和功能。方法 将H9c2细胞应用200 μmol/L的H2O2孵育0、1、3、6、12 h，应用荧光定量PCR方法检测H2O2诱导的H9c2细胞凋亡中circNFIX的表达趋势。将H9c2细胞分为空白对照组(未转染)、siRNA阴性对照组(转染siRNA阴性对照组)和siRNA组(转染siRNA)，应用荧光定量PCR方法检测siRNA对circNFIX抑制效果。将H9c2细胞分为对照组(未转染)、H2O2组(应用200 μmol/L H2O2孵育12 h)、H2O2+NC组(先转染siRNA阴性对照，余处理同H2O2组)和H2O2+siRNA组(先转染siRNA，余处理同H2O2组)，用TUNEL和Western blot方法检测敲低circNFIX对H9c2细胞凋亡的影响。结果 circNFIX表达趋势检测显示，随着H2O2孵育时间的延长，circNFIX的表达水平逐渐下降(F=23.677，P<0.01);转染siRNA可显著降低circNFIX的表达水平(F=424.70，t=24.44～25.97，P<0.01)。TUNEL和Western blot检测结果表明，转染siRNA使TUNEL阳性细胞率和Bax/Bcl-2表达水平显著降低(t=3.27～17.22，P<0.01)。结论 在H2O2诱导的H9c2细胞凋亡中circNFIX的表达水平减低，沉默circNFIX的表达抑制H9c2细胞凋亡。
[关键词] RNA，环状;过氧化氢;氧化性应激;细胞凋亡;心肌梗死
[中图分类号] R329.25 [文献标志码] A [文章编号] 2096-5532(2020)04-0389-05
doi：10.11712/jms.2096-5532.2020.56.091
[网络出版] http：//kns.cnki.net/kcms/detail/37.1517.R.20200519.1427.001.html;2020-05-20 08：53
EXPRESSION AND ROLE OF circNFIX IN H2O2-INDUCED H9C2 CELL APOPTOSIS
CUI Xianglun， JIANG Meiqing， YANG Weiwei， LIU Gege， SUN Shuqi， XU Wenhua
(Department of Inspection， The Medical Faculty of Qingdao University， Qingdao 266071， China)
[ABSTRACT]Objective To investigate the expression tendency and role of circular RNA NFIX (circNFIX) in H9c2 cell
apoptosis.Methods H9c2 cells were incubated with 200 μmol/L H2O2 for 0， 1， 3， 6， or 12 h. Quantitative real-time PCR (qPCR) was used to determine the expression tendency of circNFIX in H2O2-induced H9c2 cell apoptosis. H9c2 cells were divided into blank control group (without transfection)， siRNA negative control group (siRNA-transfected negative control group， NC group)， and siRNA group (siRNA-transfected group). qPCR assay was used to determine the inhibitory effect of siRNA on circNFIX. The H9c2 cells were divided into control group (without transfection)， H2O2 group (incubated with 200 μmol/L H2O2 for 12 h)， H2O2+NC group (first transfected with NC， followed by the same treatment as the H2O2 group)， and H2O2+siRNA group (first transfected with siRNA， followed by the same treatment as the H2O2 group). Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) and Western blot were used to determine the effect of circNFIX knockdown on H9c2 cell apoptosis. Results The testing results of circNFIX expression tendency suggested that the expression level of circNFIX was gradually down-regulated with increasing H2O2 incubation time (F=23.677，P<0.01); the expression level of circNFIX was significantly reduced after siRNA transfection (F=424.70，t=24.44-25.97，P<0.01). The results of TUNEL and Western blot indicated that siRNA transfection significantly reduced the proportion of TUNEL-positive cells and Bax/Bcl-2 levels (t=3.27-17.22，P<0.01).Conclusion In H2O2-induced H9c2 cell apoptosis， circNFIX expression is gradually down-regulated， and silencing circNFIX expression inhibits H9c2 cell apoptosis., 百拇医药(崔向伦 蒋梅青 杨维维 刘格格 孙淑琦 徐文华)
[关键词] RNA，环状;过氧化氢;氧化性应激;细胞凋亡;心肌梗死
[中图分类号] R329.25 [文献标志码] A [文章编号] 2096-5532(2020)04-0389-05
doi：10.11712/jms.2096-5532.2020.56.091
[网络出版] http：//kns.cnki.net/kcms/detail/37.1517.R.20200519.1427.001.html;2020-05-20 08：53
EXPRESSION AND ROLE OF circNFIX IN H2O2-INDUCED H9C2 CELL APOPTOSIS
CUI Xianglun， JIANG Meiqing， YANG Weiwei， LIU Gege， SUN Shuqi， XU Wenhua
(Department of Inspection， The Medical Faculty of Qingdao University， Qingdao 266071， China)
[ABSTRACT]Objective To investigate the expression tendency and role of circular RNA NFIX (circNFIX) in H9c2 cell
apoptosis.Methods H9c2 cells were incubated with 200 μmol/L H2O2 for 0， 1， 3， 6， or 12 h. Quantitative real-time PCR (qPCR) was used to determine the expression tendency of circNFIX in H2O2-induced H9c2 cell apoptosis. H9c2 cells were divided into blank control group (without transfection)， siRNA negative control group (siRNA-transfected negative control group， NC group)， and siRNA group (siRNA-transfected group). qPCR assay was used to determine the inhibitory effect of siRNA on circNFIX. The H9c2 cells were divided into control group (without transfection)， H2O2 group (incubated with 200 μmol/L H2O2 for 12 h)， H2O2+NC group (first transfected with NC， followed by the same treatment as the H2O2 group)， and H2O2+siRNA group (first transfected with siRNA， followed by the same treatment as the H2O2 group). Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) and Western blot were used to determine the effect of circNFIX knockdown on H9c2 cell apoptosis. Results The testing results of circNFIX expression tendency suggested that the expression level of circNFIX was gradually down-regulated with increasing H2O2 incubation time (F=23.677，P<0.01); the expression level of circNFIX was significantly reduced after siRNA transfection (F=424.70，t=24.44-25.97，P<0.01). The results of TUNEL and Western blot indicated that siRNA transfection significantly reduced the proportion of TUNEL-positive cells and Bax/Bcl-2 levels (t=3.27-17.22，P<0.01).Conclusion In H2O2-induced H9c2 cell apoptosis， circNFIX expression is gradually down-regulated， and silencing circNFIX expression inhibits H9c2 cell apoptosis., 百拇医药(崔向伦 蒋梅青 杨维维 刘格格 孙淑琦 徐文华)