环状RNA hsa_circ_0012779在鼻咽癌中的表达及其对细胞生物学行为影响的机制研究

Mechanism of circular RNA hsa_circ_0012779 expression in nasopharyngeal carcinoma and its influence on cell biological behavior

背景与目的:环状RNA(circular RNA,circRNA)在多种肿瘤的发展过程中发挥重要的调节作用。然而,circRNA在鼻咽癌中的异常表达和生物学功能仍不清楚。本研究旨在探讨hsa_circ_0012779对鼻咽癌细胞生物学行为的影响及其分子机制。方法:通过实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction,RTFQPCR)检测正常人鼻咽上皮细胞NP69及鼻咽癌细胞系CNE2、5-8F、HNE1、SUNE1中hsa_circ_0012779的表达。采用细胞计数试剂盒-8(cell counting kit-8,CCK-8)实验和transwell侵袭实验检测hsa_circ_0012779对鼻咽癌细胞增殖和侵袭能力的影响。采用蛋白质印迹法(Western blot)检测hsa_circ_0012779对ELAV样蛋白1(ELAV like protein 1,ELAVL1)表达的影响,通过RNA下拉实验验证hsa_circ_0012779与ELAVL1的直接结合。结果:hsa_circ_0012779在鼻咽癌组织和细胞中高表达,敲减hsa_circ_0012779能抑制鼻咽癌细胞增殖和侵袭,hsa_circ_0012779与RNA结合蛋白ELAVL1结合促进其表达并且共定位于细胞质中。同时,敲减hsa_circ_0012779对鼻咽癌细胞的影响能被ELAVL1过表达逆转。结论:hsa_circ_0012779促进ELAVL1的表达,进而促进鼻咽癌细胞增殖和侵袭,影响鼻咽癌的发生、发展。

Background and purpose:Circular RNA(circRNA)plays an important regulatory role in the development of a variety of tumors.However,the abnormal expression and biological function of circRNA in nasopharyngeal carcinoma remain unclear.This study aimed to explore the effect of hsa_circ_0012779 on the biological behavior of nasopharyngeal carcinoma cells and its molecular mechanism.Methods:The expression of hsa_circ_0012779 in human immortalized nasopharyngeal epithelial cell line NP69 and nasopharyngeal carcinoma cell lines CNE2,5-8F,HNE1 and SUNE1 was detected by real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR).Cell counting kit-8(CCK-8)assay and transwell invasion assay were used to detect the effect of hsa_circ_0012779 on the proliferation and invasion ability of nasopharyngeal carcinoma cells.The protein level of ELAV like protein 1(ELAVL1)in NPC cells with hsa_circ_0012779 knockdown was detected by Western blot.The binding of hsa_circ_0012779 and ELAVL1 was verified by RNA pull-down assay.Results:hsa_circ_0012779 was highly expressed in nasopharyngeal carcinoma tissues and cells.Knockdown hsa_circ_0012779 could inhibit the proliferation and invasion of nasopharyngeal carcinoma cells.hsa_circ_0012779 bound to RNA-binding protein ELAVL1 to promote its expression and colocalization in cytoplasm.In the meanwhile,the effect of knockdown hsa_circ_0012779 on nasopharyngeal carcinoma cells could be reversed by the overexpression of ELAVL1.Conclusion:hsa_circ_0012779 promote the expression of ELAVL1 and thus promote the proliferation and invasion of nasopharyngeal carcinoma,and influence the occurrence and development of nasopharyngeal carcinoma.