LncRNA ANRIL靶向miR-99a的表达调控鼻咽癌细胞生物学行为的影响

Effect of LncRNA ANRIL Targeting miR-99a on the Biological Behavior of Nasopharyngeal Carcinoma Cells

目的LncRNA ANRIL靶向miR-99a的表达调控鼻咽癌细胞生物学行为的影响及其机制。方法qPCR检测ANRIL和miR09a在鼻咽癌组织中的表达情况和ANTIL在不同鼻咽癌细胞株中的表达情况：分析ANRIL和鼻咽癌患者的临床病理资料之间的关联；双荧光素酶报告基因检测ANRIL与miR-99a之间的相互作用：MTF增殖实验和克隆形成实验检测抑制ANRIL后鼻咽癌细胞的增殖能力的变化情况，流式细胞术检测抑制ANRIL后对鼻咽癌细胞凋亡能力的变化情况；Western印迹检测抑制ANRIL后cyclinDl及E2F1等蛋白的表达情况：裸鼠体内成瘤实验检测抑制ANRIL后鼻咽癌细胞在裸鼠体内的成瘤情况。结果与正常组织相比，ANRIL在鼻咽癌组织中的表达水平上调，与其他鼻咽癌细胞株相比，CNEl细胞中ANRIL表达最高，miR-99a的表达水平最高；ANRIL的表达与鼻咽癌的病理分期相关以及淋巴结转移情况有关，随着分期越高，ANRIL在鼻咽癌组织中表达越高，淋巴结转移的患者中ANRIL的表达也相对较高；双荧光素酶实验证实ANRIL能与miR-99a的3’UTR特异性结合，可以调控miR-99a的表达与活性；抑制ANRIL的表达后可以抑制鼻咽癌细胞的增殖能力，同时可以促进其凋亡能力；抑制ANRIL后，cyelinDl和E2F1蛋白的激活水平相应下调，抑制ANRIL后鼻咽癌细胞在裸鼠中的生长情况受到抑制。结论ANRIL可以调控miR-99a的表达影响鼻咽癌细胞的增殖和迁移能力。

Objective To investigate the effect of LncRNA ANRIL targeting miR-99a on the biological behavior of nasopharyngeal carcinoma cells and its mechanism. Methods The expression of ANRIL and miR-99a in different nasopharyngeal carcinoma cell lines was detected by qPCR. The association between ANRIL and clinicopathological data of patients with nasopharyngeal carcinoma was analyzed. The interaction between ANRIL and miR-99a was detected by Dual-Luciferase Re- porter （DLRTM） Assay System. MTr proliferation assay and clone formation assay were used to detect the proliferation of nasopharyngeal carcinoma cells after ANRIL inhibition. The effect of ANRIL on the apoptotic ability of nasopharyngeal carcinoma cells was detected by flow cytometry. Western blotting was used to detect the expression of cyclinD1 and E2F1 after inhibiting ANRIL. The tumor formation of nude mice was detected in nude mice after inhibiting ANRIL. Results Compared with other nasopharyngeal carcinoma cell lines, the expression of ANRIL was the highest and the expres- sion level of miR-99a was the highest in CNE1 cells. The expression of ANRIL was correlated with the pathological stage of nasopharyngeal carcinoma and lymph node metastasis. The higher the ex- pression of ANRIL in nasopharyngeal carcinoma tissues, the higher the expression of ANRIL in pa- tients with lymph node metastasis. Dual-luciferase assay confirmed that ANRIL could bind to the3＇UTR of miR-99a and regulate the expression and activity of miR-99a. Inhibition of ANRIL expression can inhibit the proliferation of nasopharyngeal carcinoma cells; at the same time it promoted the apoptosis. Inhibition of ANRIL expression, cyclinD1 and E2 F1 protein expression; inhibition of ANRIL after nasopharyngeal carcinoma cells in nude mice growth was inhibited. Conclusion AN- RIL can regulate the expression of miR-99a and affect the proliferation and migration of nasopharyn- zeal carcinoma cells.