文章摘要
闫 婧,张小平,张 新,武志宏,王秀华.miR-1204表达对非小细胞肺癌细胞增殖凋亡、上皮间质转化和MAPKs信号通路的影响[J].,2022,(23):4435-4440
miR-1204表达对非小细胞肺癌细胞增殖凋亡、上皮间质转化和MAPKs信号通路的影响
Effects of miR-1204 Expression on Proliferation and Apoptosis, Epithelial Mesenchymal Transition and MAPKs Signaling Pathway in Non-Small Cell Lung Cancer Cells
投稿时间:2022-05-31  修订日期:2022-06-27
DOI:10.13241/j.cnki.pmb.2022.23.007
中文关键词: 非小细胞肺癌  miR-1204  细胞增殖  细胞凋亡  上皮间质转化
英文关键词: Non-small cell lung cancer  miR-1204  Cell proliferation  Cell apoptosis  Epithelial mesenchymal transition
基金项目:山西省基础研究计划项目(20150211068)
作者单位E-mail
闫 婧 山西医科大学附属吕梁医院呼吸与危重症医学科 山西 吕梁 033000 yanjing121314@163.com 
张小平 山西医科大学附属吕梁医院呼吸与危重症医学科 山西 吕梁 033000  
张 新 山西医科大学附属吕梁医院呼吸与危重症医学科 山西 吕梁 033000  
武志宏 山西医科大学附属吕梁医院呼吸与危重症医学科 山西 吕梁 033000  
王秀华 山西医科大学附属吕梁医院呼吸与危重症医学科 山西 吕梁 033000  
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中文摘要:
      摘要 目的:探究微小RNA-1204(miR-1204)表达对非小细胞肺癌细胞增殖凋亡、上皮间质转化(EMT)和丝裂原活化蛋白激酶(MAPKs)信号通路的影响。方法:将人非小细胞肺癌细胞A549随机分为miR-1204组(转染miR-1204mimic质粒)、NC组(转染空载质粒)和对照组(仅加转染试剂)。采用四甲基偶氮唑盐(MTT)法检测细胞增殖情况,采用流式细胞仪检测细胞凋亡情况,采用实时荧光定量聚合酶链式反应(RT-qPCR)检测细胞E-钙黏蛋白(E-cad)、N-钙黏蛋白(N-cad)和波形蛋白(Vim)mRNA的表达水平。采用RT-qPCR和蛋白免疫印迹(WB)法检测细胞p-P38、P38、p-ERK、ERK、p-JNK、JNK mRNA和蛋白的表达水平。结果:培养12、24、48 h,miR-1204组细胞增殖抑制率均高于对照组和NC组同期(P<0.05),对照组与NC组同期的细胞增殖抑制率比较差异无统计学意义(P>0.05)。但三组细胞随着培养时间延长,细胞增殖抑制率均增加,两两时间点组内比较均有差异(P<0.05)。miR-1204组细胞凋亡率高于对照组和NC组(P<0.05)。miR-1204组E-cad mRNA的表达水平高于对照组和NC组(P<0.05),N-cad、Vim mRNA的表达水平低于对照组和NC组(P<0.05)。miR-1204组p-P38、p-ERK、p-JNK mRNA和蛋白的表达水平均低于对照组和NC组(P<0.05)。结论:上调miR-1204的表达可以抑制非小细胞肺癌细胞的增殖,促进其凋亡,还可以抑制其EMT,该作用可能是通过抑制MAPKs信号通路实现的。
英文摘要:
      ABSTRACT Objective: To investigate the effect of microRNA-1204 (miR-1204) expression on proliferation and apoptosis, epithelial mesenchymal transition (EMT) and mitogen activated protein kinases (MAPKs) signaling pathway in non-small cell lung cancer cells. Methods: Human non-small cell lung cancer A549 cells were randomly divided into miR-1204 group (transfected with miR-1204mimic plasmid), NC group (transfected with empty plasmid) and control group (only with transfection reagent). Cell proliferation was detected by methyl thiazolyl tetrazolium (MTT) assay. Cell apoptosis was detected by flow cytometry, and the expression levels of E-cadherin (E-cad), N-cadherin (N-cad) and vimentin (Vim) mRNA were detected by real-time fluorescent quantitative polymerase chain reaction (RT-qPCR). The expression levels of p-P38, P38, p-ERK, ERK, p-JNK and JNK mRNA and protein were detected by RT-qPCR and Western blot (WB). Results: 12, 24, and 48 h after culture, the cell proliferation inhibition rate in the miR-1204 group was higher than that in the control group and NC group at the same period (P<0.05). There was no significant difference in the cell proliferation inhibition rate between the control group and the NC group at the same period (P>0.05). However, with the prolongation of culture time, the cell proliferation inhibition rate in the three groups increased, and there were differences between the two time points in the groups (P<0.05). The cell apoptosis rate in the miR-1204 group was higher than that in the control group and NC group (P<0.05). The expression level of E-cad mRNA in the miR-1204 group was higher than that in the control group and NC group (P<0.05), the expression levels of N-cad and Vim mRNA in the miR-1204 group were lower than those in the control group and NC group (P<0.05). The expression levels of p-P38, p-ERK and p-JNK mRNA and protein in the miR-1204 group were lower than those in control group and NC group (P<0.05). Conclusion: Up regulating the expression of miR-1204 can inhibit the proliferation and promote the apoptosis of non-small cell lung cancer cells, and also inhibit their EMT, this effect may be achieved by inhibiting MAPKs signaling pathway.
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