Wu Yun, Shen Qianwen, Chen Xiaoyu, Wu Yue, Niu Yuxu, Lv Fanzhen
Department of Thoracic Surgery, Shanghai Key Laboratory of Clinical Geriatric Medicine, Huadong Hospital Affiliated with Fudan University, Shanghai 200040, P.R. China.
Department of Radiation Oncology, Shanghai Key Laboratory of Clinical Geriatric Medicine, Huadong Hospital Affiliated with Fudan University, Shanghai 200040, P.R. China.
Oncol Lett. 2020 Dec;20(6):286. doi: 10.3892/ol.2020.12149. Epub 2020 Sep 23.
Aberrant expression of microRNAs (miRNAs or miRs) is associated with a number of human diseases, including lung cancer. Although numerous differentially expressed miRNAs have been identified in lung cancer via microarray and sequencing methods, to the best of our knowledge, only a small portion of these miRNAs have been experimentally verified. In the present study, miR-1301-3p expression levels in lung tumor tissues and lung cancer cells were measured by reverse transcription-quantitative PCR (RT-qPCR) and by analyzing previously published data. Cell Counting Kit-8 and Transwell assays were used to analyze the function of miR-1301-3p in lung cancer tissues and cells. Bioinformatics analysis, RT-qPCR, western blotting and a dual-luciferase reporter assay were performed to investigate the mechanism of miR-1301-3p in lung cancer cells. It was identified that miR-1301-3p is an upregulated miRNA in lung cancer via analyzing previously published microarray and The Cancer Genome Atlas-lung squamous cell carcinoma project data, and the upregulation of miR-1301-3p was confirmed in collected clinical samples and cells. Inhibition of miR-1301-3p suppressed lung cancer cell proliferation and migration. In addition, miR-1301-3p inhibition upregulated E-cadherin, an epithelial cell maker, and downregulated vimentin, a mesenchymal cell marker. Using bioinformatics analysis, it was revealed that polymerase I and transcript release factor (PTRF) is a target of miR-1301-3p. RT-qPCR, western blotting and dual-luciferase reporter assays demonstrated that PTRF is targeted by miR-1301-3p in lung cancer cells. The rescue experiments indicated that silencing PTRF could attenuate the inhibition of cell proliferation and migration induced by miR-1301-3p inhibitor in lung cancer cells. Furthermore, a strong negative correlation between miR-1301-3p and PTRF mRNA was identified in clinical samples. In summary, the present data highlight the involvement of miR-1301-3p in the proliferation and migration of lung cancer cells, indicating that miR-1301-3p may be a promising biomarker for lung cancer.
微小RNA（miRNA或miR）的异常表达与包括肺癌在内的多种人类疾病相关。尽管通过微阵列和测序方法已在肺癌中鉴定出众多差异表达的miRNA，但据我们所知，其中只有一小部分miRNA经过了实验验证。在本研究中，通过逆转录定量PCR（RT-qPCR）以及分析先前发表的数据，测定了肺肿瘤组织和肺癌细胞中miR-1301-3p的表达水平。使用细胞计数试剂盒-8和Transwell实验分析miR-1301-3p在肺癌组织和细胞中的功能。进行生物信息学分析、RT-qPCR、蛋白质印迹和双荧光素酶报告基因实验，以研究miR-1301-3p在肺癌细胞中的作用机制。通过分析先前发表的微阵列和癌症基因组图谱-肺鳞状细胞癌项目数据，确定miR-1301-3p是肺癌中上调的miRNA，并且在收集的临床样本和细胞中证实了miR-1301-3p的上调。抑制miR-1301-3p可抑制肺癌细胞的增殖和迁移。此外，抑制miR-1301-3p可上调上皮细胞标志物E-钙黏蛋白，并下调间充质细胞标志物波形蛋白。通过生物信息学分析发现，聚合酶I和转录释放因子（PTRF）是miR-1301-3p的靶标。RT-qPCR、蛋白质印迹和双荧光素酶报告基因实验表明，在肺癌细胞中PTRF是miR-1301-3p的靶标。拯救实验表明，沉默PTRF可减弱miR-1301-3p抑制剂对肺癌细胞增殖和迁移的抑制作用。此外，在临床样本中发现miR-1301-3p与PTRF mRNA之间存在强烈的负相关。总之，本研究数据突出了miR-1301-3p参与肺癌细胞的增殖和迁移，表明miR-1301-3p可能是一种有前景的肺癌生物标志物。
