微小RNA-203通过靶点LASP1对喉癌细胞侵袭性及凋亡的调控作用研究

Study on the effects of microRNA-203 on the invasion and apoptosis of laryngeal cancer cells via targeting LASP1

目的:探讨微小RNA-203(microRNA-203)在喉癌发生中的作用及潜在机制,阐明microRNA-203与LASP1的关系及相互作用。方法:通过实时定量聚合酶连反应(qRT-PCR)检测microRNA-203在喉癌及癌周组织中的表达。分别通过Transwell检测法和流式细胞计数检测microRNA-203对喉癌细胞的侵袭和凋亡的调控作用。双荧光素酶报告基因检测使microRNA-203与LASP1达到结合状态。通过qRT-PCR和Western blot方法检测转染microRNA-203类似物或microRNA-203抑制剂后喉癌细胞中mRNA和蛋白质水平的LASP1表达。最后通过拯救实验来检测microRNA-203是否通过其靶点LASP1来调控喉癌的进展。结果:microRNA-203在喉癌组织和细胞内有低度表达。人喉癌细胞Hep-2中敲除microRNA-203可以促进其侵袭性,并抑制其凋亡。通过双荧光素酶报告基因检测发现LASP1是microRNA-203基因的靶点。LASP1的表达由microRNA-203基因负相调节。拯救实验表明,microRNA-203基因通过其靶点LASP1对喉癌细胞的侵袭和凋亡进行调控。结论:低度表达的microRNA-203可以通过抑制LASP1来促进喉癌细胞的侵袭并抑制其凋亡。

Objective:To explore the role of microRNA-203 in laryngeal cancer and its underlying mechanism and clarify the relationship between microRNA-203 and LASP1.Method:microRNA-203 expression in laryngeal cancer tissues and paracancerous tissues was detected by quantitative real time-polymerase chain reaction(qRTPCR).The regulatory effects of microRNA-203 on invasion and apoptosis of laryngeal cancer cells were detected by Transwell assay and flow cytometry,respectively.Dual-luciferase reporter gene assay was performed to access the binding condition of microRNA-203 and LASP1.Both mRNA and protein levels of LASP1 in laryngeal cancer cells were detected after transfection with microRNA-203 mimic or microRNA-203 inhibitor by qRT-PCR and Western blot.Rescue experiments were finally performed to detect whether microRNA-203 regulates laryngeal cancer development via targeting LASP1.Result:microRNA-203 was lowly expressed in laryngeal cancer tissues and cell lines.Knockdown of microRNA-203 in Hep-2 cells can promote the invasiveness and inhibit apoptosis of laryngeal cancer cells.Subsequently,LASP1 was predicted to be the target gene of microRNA-203,which was further verified by dual-luciferase reporter gene assay.LASP1 expression was negatively regulated by microRNA-203.Furthermore,rescue experiments showed that microRNA-203 regulates invasion and apoptosis of laryngeal cancer cells via targeting LASP1.Conclusion:Low expression of microRNA-203 could promote the invasion and inhibit apoptosis of laryngeal cancer cells viainhibiting LASP1.microRNA-203 and LASP1 both play a very important role in the development of laryngeal cancer.