长链非编码RNA SPRY4-IT1在食管鳞癌中的表达及对细胞生长的影响*

摘要

Objective:This study aimed to clarify the correlation of SPRY4-IT1 expression with the clinicopathological character-istics and prognosis of patients with esophageal squamous cell carcinoma (ESCC), as well as the role of SPRY4-IT1 in promoting ES-CC cell growth. Methods:Quantitative real-time polymerase chain reaction for SPRY4-IT1 expression was performed on 50 paired can-cerous and adjacent non-cancerous esophageal specimens. Small interfering RNA was used to suppress SPRY4-IT1 expression to fur-ther explore its role in tumor progression. Cell viability was tested in vitro by MTT assay (OD=490 nm), and cell apoptosis and cell cy-cle were investigated by flow cytometry. Results:We found markedly elevated SPRY4-IT1 expression in cancerous tissues compared with adjacent non-cancerous tissues (90%, P<0.01). Relative SPRY4-IT1 expression levels were correlated with some clinicopathologi-cal characteristics, such as tumor size (χ2=5.333, P=0.021), elevated TNM (2009) stage classi fi cation (χ2=5.556, P=0.018), and de-creased overall survival rates (χ2=5.296, P=0.021). SPRY4-IT1 expression level was not correlated with patient age, gender, smoking status, or alcohol consumption (all P>0.05). Further experiments showed that SPRY4-IT1 expression levels were significantly higher in three ESCC cell lines than in the normal human esophageal epithelial cell line Het-1A. In vitro assays of the ESCC cell line KYSE30 demonstrated that knockdown of SPRY4-IT1 expression by small interfering RNA reduced cell growth, mediated cell cycle arrest at the G0-G1 phase, and promoted cell apoptosis (all P<0.01). Conclusion:SPRY4-IT1 was overexpressed in ESCC tissues and ESCC cell lines and promoted the growth of ESCC cells. The dysregulated expression of long non-coding RNA SPRY4-IT1 may play an important role in the process of ESCC development and may be developed as a useful biomarker for the diagnosis and prognosis of ESCC.%目的：探讨长链非编码RNA SPRY4-IT1与食管鳞状细胞癌（ESCC）的临床病理及预后的相关性，以及对细胞生长的影响。方法：收集2008年1月至2009年12月南京医科大学附属南京医院肿瘤外科50例ESCC手术切除标本（包括癌组织和癌旁组织），荧光实时定量PCR（qRT-PCR）检测50例ESCC中SPRY4-IT1的表达情况，分析其与临床病理及预后的关系，用小干扰RNA（siRNA）干扰SPRY4-IT1后MTT法检测其对细胞生长的影响，流式细胞检测对细胞凋亡和周期的影响。结果：45例（90%）标本中SPRY4-IT1呈阳性表达，SPRY4-IT1在癌组织中的表达量显著高于癌旁组织（t=5.377，P<0.01）。SPRY4-IT1的相对表达水平与肿瘤大小、临床分期相关（P均<0.05）。SPRY4-IT1在ESCC细胞株中表达量高于正常食管上皮细胞。KYSE30细胞中干扰SPRY4-IT1的表达后可明显减慢细胞生长，阻滞细胞周期并促进细胞凋亡（P<0.01）。结论：SPRY4-IT1在ESCC组织中显著高表达，并且能促进ESCC细胞生长，可能成为ESCC诊断和判断预后的重要分子标记物。