摘要
目的研究长链非编码RNA(lncRNA)DHRS4-AS1对微小RNA(miR)-221的靶向调控及对肺癌细胞增殖、迁移、侵袭、凋亡的影响。方法实时荧光定量PCR(qPCR)检测肺癌组织中DHRS4-AS1和miR-221表达。在肺癌细胞A549中转染pcDNA3.1-DHRS4-AS1,噻唑蓝(MTT)和平板克隆实验检测细胞存活与克隆形成,流式细胞术检测细胞凋亡,Transwell小室法检测细胞迁移和侵袭,免疫印迹试验检测P21、E-钙黏蛋白(E-cadherin)、Caspase-3、基质金属蛋白酶-2(MMP-2)蛋白表达。双萤光素酶报告实验验证DHRS4-AS1与miR-221靶向。pcDNA3.1-DHRS4-AS1和miR-221共转染,观察过表达miR-221对DHRS4-AS1诱导的细胞A549增殖、凋亡、迁移、侵袭的影响。结果与癌旁组织比较,肺癌组织中DHRS4-AS1表达量明显减少,miR-221表达量显著增加(P<0.05)。过表达DHRS4-AS1增加凋亡率、P21、Caspase-3、E-cadherin蛋白表达,降低细胞存活率、MMP-2蛋白表达、克隆形成数、迁移细胞数、侵袭细胞数(P<0.05)。DHRS4-AS1靶向、调控miR-221。过表达miR-221能逆转DHRS4-AS1对细胞A549增殖、迁移、侵袭、凋亡的作用。结论DHRS4-AS1可靶向调控miR-221抑制肺癌细胞增殖、迁移和侵袭,诱导细胞凋亡。
Objective To investigate the targeting regulation of long non-coding RNA(lncRNA)DHRS4-AS1 on microRNA(miR)-221 and the proliferation,migration,invasion and apoptosis of lung cancer cells.Methods Reverse transcription quantitative real-time polymerase chain reaction(RT-qPCR)was used to detect the expressions of DHRS4-AS1 and miR-221 in lung cancer tissues.After transfecting with pcDNA3.1-DHRS4-AS1 or pcDNA3.1-NC into lung cancer cells A549,cell survival,colony formation and apoptosis were detected by MTT assay,colony formation assay and flow cytometry,respectively.Cell migration and invasion were examined by Transwell assay.Protein expressions of P21,E-c adherin,Caspase-3 and matrix metalloproteinase-2(MMP-2)were detected by Western blot.Dual luciferase reporting assay was performed to validate the targeting relationship between DHRS4-AS1 and miR-221.After co-transfecting pcDNA3.1-DHRS4-AS1 and miR-221 overexpression plasmid,the effects of overexpression of miR-221 on proliferation,apoptosis,migration and invasion of A549 cells induced by DHRS4-AS1 were observed.Results Compared with the adjacent normal tissue,DHRS4-AS1 was significantly downregulated and miR-221 was significantly upregulated in lung cancer tissue(P<0.05).Overexpression of DHRS4-AS1 significantly increased apoptotic rate and protein levels of P21,Caspase-3 and E-cadherin,but decreased survival rate,protein level of MMP-2 and numbers of colonies,migratory cells and invasive cells P<0.05).Overexpression of miR-221 could reverse the effects of DHRS4-AS1 on the proliferation,migration,invasion and apoptosis of A549 cells.Conclusion DHRS4-AS1 inhibits the proliferation,migration and invasion of lung cancer cells and induces cell apoptosis by targeting miR-221.
作者
马强
谌登珍
MA Qiang;CHEN Dengzhen(Department of Respiratory and Critical Care Medicine,Shangluo Central Hospital,Shaanxi,Shangluo 726000,China;不详)
出处
《河北医药》
2025年第6期925-929,共5页
Hebei Medical Journal
基金
陕西省重点研发计划项目(编号:2020SF-116)。
