摘要
目的探讨长链非编码RNA HOTAIR在非小细胞肺癌(NSCLC)组织及4株细胞系(A549、SPC-A1、SK-MES-1和16HBE)中的表达,并分析其对细胞侵袭和迁移能力的影响。方法通过定量反转录PCR(qRT-PCR)检测HOTAIR在38例NSCLC组织及4株细胞系中的表达水平,并进一步利用过表达和RNA干扰技术探讨HOTAIR的生物学功能。通过分别转染pcDNA-HOTAIR或si-HOTAIR来上调或下调HOTAIR的表达,以空载体(pcDNA3.1-NC)和阴性对照(si-NC)作为对照组,用qRT-PCR检测转染效率。用MTT法和Transwell法评估异常表达的HOTAIR对NSCLC细胞增殖、迁移和侵袭能力的影响。结果 38例NSCLC组织中HOTAIR相对表达水平为24.48±59.55。与正常支气管上皮细胞系16HBE相比,HOTAIR在SPCA1和SK-MES-1细胞中相对高表达,而在A549细胞中相对低表达。转染HOTAIR siRNA 48h后,A549和SPC-A1细胞中HOTAIR表达下调;转染pcDNA3.1-HOTAIR 48h后,A549细胞中HOTAIR表达上调。MTT实验显示,通过RNAi技术来抑制HOTAIR的表达,对NSCLC细胞的增殖能力无明显影响。Transwell实验显示,通过转染si-HOTAIR来下调HOTAIR表达可抑制癌细胞的迁移和侵袭(P<0.05);相反,过表达HOTAIR可以显著促进癌细胞的迁移和侵袭(P<0.05)。结论 HOTAIR在NSCLC中异常高表达,能显著增强NSCLC细胞的迁移和侵袭能力,提示可能与不良预后相关。
Objective To investigate the expression profile of long noncoding RNA HOTAIR in non-small cell lung cancer （NSCLC） tissues and four different cell lines （A549, SPC-A1, SK-MES-1 and 16HBE）, and to study its biological functions in NSCLC carcinogenesis and progression, especially on cell migration and invasion. Methods Quantitative reverse-transcription PCR （qRT- PCR） was performed to detect the relative expression of HOTAIR in thirty-eight NSCLC tissues and four cell lines（ A549, SPC-A1, SK-MES-1 and 16HBE）. To further explore its biological function, techniques of overexpression and RNA interference（RNAi） were applied, pcDNA-HOTAIR or si-HOTAIR was transfected to up or down-regulate HOTAIR expression in NSCLC cells respectively, and the transfection efficiency was evaluated by qRT-PCR. MTT assay and Transwell assay were performed to evaluate the effect of ectopic HOTAIR expression on proliferation, migration and invasion potential of NSCLC cells. Results The relative expression of HOTAIR in 38 cases of NSCLC tissues was 24. 48~59. 55. Compared with normal bronchial epithelium cell 16HBE, there was a relatively high ex- pression of HOTAIR in SPC-A1 and SK-MES-1 cells, and a relatively low expression in A549 cell. The HOTAIR expression was down- regulated in A549 and SPC-A1 cells at 48h after transfection of HOTAIR siRNA, and up-regulated in A549 cells at 48h after transfection of pcDNA3.1-HOTAIR. MTT assay indicated that RNAi-mediated suppression of HOTAIR had little effect on cell proliferation,while Transwell assay indicated that inhibition of HOTAIR by si-HOTAIR could repress cell migration and invasion （P〈 0.05） ;con- versely, overexpression of HOTAIR could significantly promote cell migration and invasion（P〈0. 05）. Conclusion The expression of HOTAIR is significantly up-regulated in NSCLC, which can promote cell migration and invasion. HOTAIR may be a new biomarker of poor prognosis in NSCLC.
出处
《临床肿瘤学杂志》
CAS
2014年第8期684-689,共6页
Chinese Clinical Oncology
基金
国家自然科学基金资助项目(81070620)
