长链非编码RNA HAGLROS在食管鳞癌组织中的表达及其对食管鳞癌细胞生物学行为的影响

Expression and biological function of lncRNA HAGLROS in the esophageal squamous cell carcinoma

目的探讨长链非编码RNA HAGLROS在食管鳞癌组织中的表达及其与食管鳞癌患者临床病理参数之间的关系,分析HAGLROS对食管鳞癌细胞增殖及转移的影响。方法运用逆转录-聚合酶链反应(RT-PCR)检测HAGLROS在42对食管鳞癌组织与癌旁食管正常组织及食管上皮细胞Het1A及食管鳞癌细胞EC109和KYSE30中的表达。运用小干扰RNA(siRNA)技术在食管鳞癌细胞系中抑制HAGLROS的内源性表达,RT-PCR检测沉默效率。采用CCK-8及Transwell法分析敲减HAGLROS后对食管鳞癌细胞体外增殖及迁移能力的影响。结果食管鳞癌组织中HAGLROS的表达明显高于癌旁正常食管组织,差异有统计学意义(t=2.913,P=0.006)。HAGLROS的表达与食管鳞癌患者的淋巴结转移及TNM分期呈现明显的正相关(t=2.866,P=0.007;t=3.882,P=0.001);HAGLROS的表达与食管鳞癌患者的组织学分化程度无明显相关性(t=1.156,P=0.254)。HAGLROS在KYSE30与EC109细胞中表达明显高于在食管上皮细胞系Het-1A的表达(t=8.384,P=0.001;t=6.767,P=0.003)。在EC109与KYSE30细胞中瞬时转染针对HAGLROS的siRNA,能够有效抑制HAGLROS的表达。CCK-8细胞活性实验的结果表明,2种食管鳞癌细胞在转染siHAGLROS的96 h后的增殖速率与对照组相比明显降低。Transwell实验结果显示,敲减HAGLROS后,2种食管鳞癌细胞的迁移能力与对照组相比明显下降。结论HAGLROS在食管鳞癌组织中表达明显增加,与食管鳞癌患者的淋巴结转移及TNM分期具有较强的相关性。HAGLROS在食管鳞癌细胞系中表达较高,敲减HAGLROS后,食管鳞癌细胞的增殖及迁移能力均受到明显的抑制,提示HAGLROS在食管鳞癌的发生、发展中起到促癌的作用,有望成为食管鳞癌靶向治疗的新靶点。

Objective To investigate the expression of long non-coding RNA HAGLROS in the esophageal squamous cell carcinoma tissues and its correlations with the clinicopathological characteristics of esophageal squamous cell carcinoma patients,and to analyze the effects of HAGLROS on the proliferation and migration of esophageal squamous cell carcinoma cells.Methods Reverse transcription polymerase chain reaction(RT-PCR)was used to examine the expression of HAGLROS in forty-two cases of esophageal squamous cell carcinoma tissues and their corresponding non-tumoral esophagus tissues,and the expression of HAGLROS in the esophageal epithelial cell Het1A,esophageal squamous cell carcinoma cell lines EC109 and KYSE30.Small-interfering RNA(siRNA)technology was utilized to inhibit the endogenous expression of HAGLROS in the esophageal squamous cell carcinoma cell lines,and RT-PCR was conducted to confirm the inhibition efficiencies.CCK-8 and transwell experiments were performed to determine the role of HAGLROS in the esophageal squamous cell carcinoma cell proliferation and migration.Results HAGLROS expression was significantly elevated in the esophageal squamous cell carcinoma tissues compared to the matched normal esophageal tissues(t=2.913,P=0.006).There was no relationship between HAGLROS expression and the differentiation grade of esophageal squamous cell carcinoma patients(t=1.156,P=0.254),while HAGLROS level was positively correlated to the lymph node metastasis and TNM stage of esophageal squamous cell carcinoma patients(t=2.866,P=0.007;t=3.882,P=0.001).Besides,the expression of HAGLROS was higher in the YSE30 and EC109 cells than the normal esophagus epithelial cells Het-1A(t=8.384,P=0.001;t=6.767,P=0.003).SiRNAs targeting HAGLROS were transfected into EC109 and KYSE30 cells and obviously inhibited HAGLROS expression in the esophageal squamous cell carcinoma cells.Silencing of HAGLROS decreased two kinds of esophageal squamous cell carcinoma cells proliferation at 96 h,as evidenced by CCK-8 assays.Transwell experiments demonstrated that knockdown of HAGLROS weakened the migration abilities of two kinds of esophageal squamous cell carcinoma cells.Conclusion HAGLROS expression was upregulated in the esophageal squamous cell carcinoma tissues and correlated with the lymph node metastasis and advanced TNM stage of esophageal squamous cell carcinoma patients.HAGLROS was elevated in esophageal squamous cell carcinoma cell lines and inhibition of HAGLROS repressed cell growth and metastasis in vitro.Overall,these results indicated that HAGLROS plays a critical role in esophageal squamous cell carcinoma progression and may serve as a novel target for esophageal squamous cell carcinoma treatment.