摘要
目的观察微小RNA(miRNA,miR)-148a-3p与溶质载体家族7成员11(SLC7A11)在肺癌组织的表达及其对肿瘤细胞铁死亡的影响。方法选取河南省人民医院胸外科2020年6月至2023年6月收治的肺癌临床样本作为研究对象,癌旁组织作为对照研究对象。采用荧光定量聚合酶链反应(PCR)和蛋白质免疫印迹计数分析miR-148a-3p与SLC7A11的表达水平。人非小细胞肺癌细胞H1299分为miRNA对照组和miR-148a-3p组,采用慢病毒感染构建细胞系。采用细胞计数试剂盒(CCK-8)和克隆形成实验分析两组细胞的增殖能力;采用体外移植瘤分析两组细胞在裸鼠体内的体积和重量;采用蛋白质免疫印迹分析铁死亡相关蛋白谷胱甘肽过氧化物酶4表达水平;生物信息学和双荧光素酶报告基因分析miR-148a-3p靶基因,并分析靶基因表达水平。组间计量数据比较采用t检验。结果癌旁组织中miR-148a-3p表达水平(1.02±0.19)明显高于肺癌组织(0.54±0.19),差异有统计学意义(t=14.790,P<0.05)。癌旁组织中SLC7A11信使RNA(mRNA)表达水平(0.66±0.13)明显低于肺癌组织(1.48±0.17),差异有统计学意义(t=27.180,P<0.05)。miRNA对照组细胞吸光度(A)值(1.98±0.11)明显高于miR-148a-3p组细胞(1.48±0.19),差异有统计学意义(t=5.674,P<0.05)。miRNA对照组细胞克隆形成率[(85.07±4.50)%]明显高于miR-148a-3p组细胞[(53.68±8.89)%],差异有统计学意义(t=7.714,P<0.05)。miRNA对照组细胞肿瘤体积[(888.40±90.22)mm^(3)]明显高于miR-148a-3p组细胞[(609.78±102.92)mm^(3)],差异有统计学意义(t=6.438,P<0.05)。miRNA对照组细胞肿瘤重量[(5.34±0.75)g]明显高于miR-148a-3p组细胞[(2.28±0.71)g],差异有统计学意义(t=9.379,P<0.05)。SLC7A11是miR-148a-3p的靶基因。miRNA对照组细胞SLC7A11(0.78±0.07)明显高于miR-148a-3p组细胞(0.48±0.09),差异有统计学意义(t=7.580,P<0.05)。miRNA对照组细胞谷胱甘肽过氧化物酶4(1.09±0.09)明显高于miR-148a-3p组细胞(0.39±0.09),差异有统计学意义(t=15.480,P<0.05)。结论miR-148a-3p在肺癌组织中表达显著下调,通过SLC7A11调节肺癌细胞铁死亡,进而影响肺癌的生长和发展。
Objective To observe the expression of microRNA(miR)-148a-3p and solute carrier family 7 member 11(SLC7A11)in lung cancer tissues and their effects on ferroptosis of tumor cells.Methods Clinical samples of lung cancer patients admitted to our hospital from June 2020 to June 2023 were selected as the research objects,and adjacent tissues were selected as the control objects.The expression levels of miR-148a-3p and SLC7A11 were analyzed by fluorescence quantitative polymerase chain reaction(PCR)and Western blotting.Human non-small cell lung cancer cell line H1299 was divided into miRNA control group and miR-148a-3p group,and the cell line was constructed by lentiviral infection.The proliferation ability of the two groups was analyzed by cell counting kit-8(CCK-8)and colony formation experiment.The volume and weight of the two groups in nude mice were analyzed by in vitro xenograft.The expression level of ferroptosis-related protein glutathione peroxidase 4 was analyzed by Western blotting.The target gene of miR-148a-3p was analyzed by bioinformatics and dual luciferase reporter gene analysis,and the expression level of the target gene was analyzed.The t test was used for comparison of measurement data between groups.Results The expression level of miR-148a-3p in adjacent tissues(1.02±0.19)was significantly higher than that in lung cancer tissues(0.54±0.19,t=14.790,P<0.05).The mRNA expression level of SLC7A11 carrier family 7 member 11 in adjacent tissues(0.66±0.13)was significantly lower than that in lung cancer tissues(1.48±0.17,t=27.180,P<0.05).The absorbance(A)of the miRNA control group(1.98±0.11)was significantly higher than that of the miR-148a-3p group(1.48±0.19,t=5.674,P<0.05).The colony formation rate of the miRNA control group[(85.07±4.50)%]was significantly higher than that of the miR-148a-3p group[(53.68±8.89)%,t=7.714,P<0.05].The tumor volume of the miRNA control group[(888.40±90.22)mm^(3)]was significantly greater than that of the miR-148a-3p group[(609.78±102.92)mm^(3),t=6.438,P<0.05].The tumor weight of the miRNA control group[(5.34±0.75)g]was significantly greater than that of the miR-148a-3p group[(2.28±0.71)g,t=9.379,P<0.05].SLC7A11 is the target gene of miR-148a-3p.The SLC7A11 of the miRNA control group(0.78±0.07)was significantly higher than that of the miR-148a-3p group(0.48±0.09,t=7.580,P<0.05).The glutathione peroxidase 4 of the miRNA control group(1.09±0.09)was significantly higher than that of the miR-148a-3p group(0.39±0.09,t=15.480,P<0.05).Conclusion The expression of miR-148a-3p is significantly down-regulated in lung cancer tissues,and it regulates ferroptosis of lung cancer cells through SLC7A11 and affects the growth and development of lung cancer.
作者
魏文学
张全
黄锵文
魏立
Wei Wenxue;Zhang Quan;Huang Qiangwen;Wei Li(Department of Thoracic Surgery,Henan Provincial People’s Hospital,People’s Hospital of Zhengzhou University,School of Clinical Medicine,Henan University,Zhengzhou 450003,China)
出处
《中华实验外科杂志》
CAS
2024年第4期751-754,共4页
Chinese Journal of Experimental Surgery
作者简介
通信作者:魏立,Email:546696594@qq.com。
