Roa Wilson, Brunet Bryan, Guo Linghong, Amanie John, Fairchild Alysa, Gabos Zsolt, Nijjar Tirath, Scrimger Rufus, Yee Don, Xing James
Alberta Laboratory for Environmental Cancer Risk & Assessment, Alberta, Canada.
Clin Invest Med. 2010 Apr 1;33(2):E124. doi: 10.25011/cim.v33i2.12351.
Small non-coding microRNAs (miRNAs) are key components of cancer development and are considered as potential biomarkers for cancer diagnosis and treatment monitoring. This study investigated miRNA expression profiles of human cancer cells in order to develop a screening method for lung cancer.
A series of lung cancer related miRNAs (miR-21, miR-145, miR-155, miR-205, miR-210, miR-92, miR-17-5p, miR-143, miR-182, miR-372, let-7a) were selected as candidates for miRNA expression profiles of human lung cancer cell lines (A549, SK-mes-1). MicroRNA u6 was the endogenous control. Cancer cell lines for positive controls; breast MCF-7, prostate Du-145, and glioblastoma U118. The negative control was normal lung fibroblast cell line MRC-5. RT-PCR was performed on StepOnePlus (Applied Biosystem, USA). MiRNA expressions of malignant cells were compared with normal fibroblast cells as well as endogenous control (u6) using the thermal cycle at threshold. Assessment of miRNA expression profiles were then performed using agglomerative hierarchical cluster analysis software (SPSS13, USA).
We demonstrated that miR-21, miR-182 and let7-5a were over-expressed, and miR-145 and miR-155 were under-expressed in all cancer cell lines. Combined with the cluster analysis we were able to clearly distinguish cell lines for normal fibroblasts, breast cancer, prostate cancer, glioblastoma, and lung cancer.
There is potential utility of screening for lung cancer with miRNA expression profiles. Future work will focus on the sensitivity of such miRNA expression profiles in screening sputum for lung cancer, which can be performed in real time.
小型非编码微小RNA（miRNA）是癌症发展的关键组成部分，被视为癌症诊断和治疗监测的潜在生物标志物。本研究调查了人类癌细胞的miRNA表达谱，以开发一种肺癌筛查方法。
选择一系列与肺癌相关的miRNA（miR-21、miR-145、miR-155、miR-205、miR-210、miR-92、miR-17-5p、miR-143、miR-182、miR-372、let-7a）作为人类肺癌细胞系（A549、SK-mes-1）miRNA表达谱的候选物。微小RNA u6作为内参。癌细胞系作为阳性对照；乳腺癌MCF-7、前列腺癌Du-145和成胶质细胞瘤U118。阴性对照为正常肺成纤维细胞系MRC-5。在StepOnePlus（美国应用生物系统公司）上进行逆转录聚合酶链反应（RT-PCR）。使用阈值处的热循环将恶性细胞的miRNA表达与正常成纤维细胞以及内参（u6）进行比较。然后使用凝聚层次聚类分析软件（美国SPSS13）进行miRNA表达谱评估。
我们证明，在所有癌细胞系中miR-21、miR-182和let7-5a过表达，而miR-145和miR-155表达不足。结合聚类分析，我们能够清楚地区分正常成纤维细胞、乳腺癌、前列腺癌、成胶质细胞瘤和肺癌的细胞系。
利用miRNA表达谱筛查肺癌具有潜在的实用性。未来的工作将集中于这种miRNA表达谱在肺癌痰筛查中的敏感性，这可以实时进行。
