Su Chongyu, Shi Kang, Cheng Xu, Han Yi, Li Yunsong, Yu Daping, Liu Zhidong
1 Department of Thoracic Surgery, Beijing Chest Hospital, Beijing Tuberculosis and Thoracic Tumor Research Institute, Capital Medical University, Beijing, China.
Technol Cancer Res Treat. 2018 Jan 1;17:1533033818790490. doi: 10.1177/1533033818790490.
We aimed to detect the role of LINC00472 via regulating miR-24-3p and death effector domain-containing DNA-binding protein in lung adenocarcinoma.
Long noncoding RNA, microRNA, and messenger RNA levels were determined using reverse transcription quantitative polymerase chain reaction. The expression of death effector domain-containing DNA-binding protein was determined using Western blot assay. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and colony formation assay were conducted to explore the proliferation of cells. The cell apoptosis was tested by flow cytometry assay. Target relationships between miR-24-3p, death effector domain-containing DNA-binding protein, and LINC00472 were validated by dual-luciferase reporter gene assay.
LINC00472 and death effector domain-containing DNA-binding protein were found to be underexpressed, whereas miR-24-3p was found overexpressed in lung adenocarcinoma cell lines and tissues. Both LINC00472 and death effector domain-containing DNA-binding protein can bind to miR-24-3p. Overexpression of LINC00472 led to higher death effector domain-containing DNA-binding protein level, demoting cell proliferation while promoting apoptosis. Overexpression of miR-24-3p reduced death effector domain-containing DNA-binding protein level, which facilitated cell proliferation and inhibited cell apoptosis, as well as to some extent restrained the effects of LINC00472. The high expression of miR-24-3p in tumor cells was negatively related to LINC00472 and death effector domain-containing DNA-binding protein, whereas the expression of LINC00472 and that of death effector domain-containing DNA-binding protein were positively correlated.
Our findings suggested that LINC00472 contributed to the increase in lung adenocarcinoma cell apoptosis and the inhibition of proliferation via regulating miR-24-3p/ DEDD, which might provide a novel insight into potential therapeutic approach for lung adenocarcinoma.
我们旨在通过调控miR-24-3p和含死亡效应结构域的DNA结合蛋白来检测LINC00472在肺腺癌中的作用。
使用逆转录定量聚合酶链反应测定长链非编码RNA、微小RNA和信使RNA水平。使用蛋白质免疫印迹法测定含死亡效应结构域的DNA结合蛋白的表达。进行3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法和集落形成试验以探究细胞增殖情况。通过流式细胞术检测细胞凋亡。通过双荧光素酶报告基因试验验证miR-24-3p、含死亡效应结构域的DNA结合蛋白和LINC00472之间的靶向关系。
发现LINC00472和含死亡效应结构域的DNA结合蛋白在肺腺癌细胞系和组织中表达下调，而miR-24-3p表达上调。LINC00472和含死亡效应结构域的DNA结合蛋白均可与miR-24-3p结合。LINC00472的过表达导致含死亡效应结构域的DNA结合蛋白水平升高，降低细胞增殖同时促进细胞凋亡。miR-24-3p的过表达降低了含死亡效应结构域的DNA结合蛋白水平，促进细胞增殖并抑制细胞凋亡，并且在一定程度上抑制了LINC00472的作用。肿瘤细胞中miR-24-3p的高表达与LINC00472和含死亡效应结构域的DNA结合蛋白呈负相关，而LINC00472和含死亡效应结构域的DNA结合蛋白的表达呈正相关。
我们的研究结果表明，LINC00472通过调控miR-24-3p/DEDD促进肺腺癌细胞凋亡并抑制增殖，这可能为肺腺癌的潜在治疗方法提供新的见解。
