摘要
目的 探讨miR-558对肺癌细胞增殖、迁移及凋亡的影响。方法 收集医院2020年5月至8月收治的35例肺癌患者的癌组织及其癌旁组织样本,并取对数生长期的人肺癌细胞A549、人支气管上皮细胞BEAS-2B,采用实时荧光定量聚合酶链反应(qPCR)法检测circ_0001017和miR-558表达水平。取对数生长期的A549细胞进行转染,并分为空质粒(pcDNA)组(A组)、pcDNA-circ_0001017组(B组)、pcDNA-circ_0001017+miR-NC组(C组)、pcDNA-circ_0001017+miR-558组(D组)。采用双荧光素酶报告实验检测circ_0001017与miR-558的靶向关系;采用四甲基偶氮唑盐(MTT)法、平板克隆形成实验、Transwell实验、流式细胞术分别检测细胞的活性、克隆、迁移、凋亡情况;采用qPCR法检测circ_0001017、miR-558表达水平;采用免疫印迹(Westernblot)法检测B细胞淋巴瘤因子-2蛋白(Bcl-2)和Bcl-2相关X蛋白(Bax)的表达水平。结果 与癌旁组织比较,肺癌组织中circ_0001017的表达水平显著降低(P<0.05),miR-558的表达水平显著升高(P<0.05);与BEAS-2B细胞比较,A549细胞中circ_0001017的表达水平显著降低(P<0.05),miR-558的表达水平显著升高(P<0.05)。miR-558过表达可显著降低野生型载体(WT)-circ_0001017的荧光素酶活性(P<0.05)。与A组比较,B组细胞的增殖抑制率、凋亡率、circ_0001017和Bax表达水平均显著升高(P<0.05),细胞克隆形成数、迁移细胞数均显著减少(P<0.05),miR-558和Bcl-2表达水平均显著降低(P<0.05);与C组比较,D组细胞的增殖抑制率、凋亡率、circ_0001017和Bax表达水平均显著降低(P<0.05),细胞克隆形成数、迁移细胞数均显著增多(P<0.05),Bcl-2和miR-558表达水平均显著升高(P<0.05)。结论 过表达miR-558会减弱过表达circ_0001017对肺癌细胞的增殖、克隆形成、迁移的抑制作用及促细胞凋亡作用。
Objective To investigate the effect of miR-558 on the proliferation,migration and apoptosis of lung cancer cells.Methods The cancer tissue and adjacent tissue specimens of 35 patients with lung cancer admitted to the hospital from May to August 2020 were collected.The human lung cancer cells A549 and human bronchial epithelial cells BEAS-2B in logarithmic growth phase were selected,to detect the expression levels of circ_0001017 and miR-558 by the real-time fluorescence quantitative polymerase chain reaction(qPCR)method.The A549 cells in logarithmic growth phase were selected for transfection and divided into the empty plasmid(pcDNA)group(group A),pcDNA-circ_0001017 group(group B),pcDNA-circ_0001017+miR-NC group(group C),and pcDNA-circ_0001017+miR-558 group(group D).The dual-luciferase reporter assay was used to detect the targeting relationship between circ_0001017 and miR-558.The MTT assay,plate clone formation assay,Transwell assay and flow cytometry were used to detect the cell activity,cloning,migration,and apoptosis,respectively.The qPCR method was used to detect the expression levels of circ_0001017 and miR-558.The Western blot was used to detect the expression levels of B-cell lymphoma-2 protein(Bcl-2)and Bcl-2-associated X protein(Bax).Results Compared with those in the adjacent tissues,the expression level of circ_0001017 in the lung cancer tissue significantly decreased(P<0.05),while the expression level of miR-558 significantly increased(P<0.05).Compared with those in the BEAS-2B cells,the expression level of circ_0001017 in A549 cells significantly decreased(P<0.05),while the expression level of miR-558 significantly increased(P<0.05).Overexpression of miR-558 could significantly decrease the luciferase activity of wild-type vector(WT)-circ_0001017(P<0.05).Compared with those in group A,the proliferation inhibition rate,the apoptosis rate,and the expression levels of circ_0001017 and Bax in group B significantly increased(P<0.05),while the number of cell clone formation and cell migration significant
作者
郭淑娟
郑成玲
高晓盼
孙晓彤
GUO Shujuan;ZHENG Chenging;GAO Xiaopan;SUN Xiaotong(Sunshine Union Hospital,Weifang,Shandong,China 261000;Department of Anesthesiology,Weifang Medical College,Weifang,Shandong,China 261000)
出处
《中国药业》
CAS
2024年第11期57-61,共5页
China Pharmaceuticals
基金
山东省自然科学基金[ZR2019PH037]。
