长链非编码RNA SNHG16 靶向微小RNA-7-5p对喉癌细胞增殖、迁移、侵袭的影响

The effect of SNHG16 regulate on proliferation,migration and invasion of laryngeal carcinoma cells by targeting miR-7-5p

目的探讨长链非编码RNA SNHG16对喉癌细胞增殖、迁移、侵袭的影响及其作用机制。方法选取2016年10月至2018年12月重庆医科大学附属第三医院收治的25例喉癌病人,于术中切取喉癌组织及其相应癌旁组织并冻存。体外培养喉癌Hep-2细胞,Hep-2细胞,按照随机数字表法分为si-NC组(转染无意义干扰序列si-NC)、si-SNHG16组(转染si-SNHG16)、si-SNHG16+anti-miR-NC组(共转染si-SNHG16与anti-miR-NC)、si-SNHG16+anti-miR-7-5p组(共转染si-SNHG16与anti-miR-7-5p)。采用实时荧光定量PCR(qRT-PCR)检测喉癌及其癌旁组织中SNHG16、微小RNA-7-5p(miR-7-5p)的表达。采用四甲基偶氮唑盐微量酶反应比色法(MTT法)、Transwell实验检测抑制SNHG16表达后Hep-2细胞的增殖、迁移及侵袭能力的变化。生物信息学预测SNHG16的靶标miRNA,双荧光素酶报告实验验证SNHG16与miR-7-5p的靶向作用。抑制miR-7-5p表达验证SNHG16对Hep-2细胞增殖、迁移、侵袭的作用。结果喉癌组织中SNHG16的表达水平较癌旁组织升高(P<0.05),miR-7-5p的表达水平明显降低(P<0.05);与si-NC组相比,si-SNHG16组抑制SNHG16的表达后可明显抑制Hep-2细胞的增殖能力[48 h:(0.95±0.09)比(0.47±0.05),P<0.05],迁移[(124±12)比(51±5),P<0.05]及侵袭细胞数[(115±11)比(43±4),P<0.05]减少;SNHG16可靶向结合miR-7-5p;抑制miR-7-5p可逆转抑制SNHG16对Hep-2细胞增殖、迁移、侵袭的抑制作用。结论抑制SN⁃HG16表达可负性调控miR-7-5p的表达而减弱喉癌细胞增殖、迁移及侵袭能力。

Objective To investigate the effects of long non-coding RNA SNHG16(LncRNA SNHG16)on proliferation,migration and invasion of laryngeal carcinoma cells and its mechanism.Methods Twenty-five patients with laryngeal cancer admitted to the Third Affiliated Hospital of Chongqing Medical University from October 2016 to December 2018 were selected,and the laryngeal can⁃cer tissue and its corresponding adjacent tissues were cut and frozen during the operation.Laryngeal carcinoma Hep-2 cells and Hep-2 cells were cultured in vitro,and they were randomly divided into si-NC group(transfected with meaningless interference sequence si-NC),si-SNHG16 group(transfected with si-SNHG16),and si-SNHG16+anti-miR-NC group(co-transfection si-SNHG16 and anti-miRNC),si-SNHG16+anti-miR-7-5p group(co-transfection si-SNHG16 and anti-miR-7-5p).The expression of SNHG16 and miR-7-5p in laryngeal carcinoma tissues was detected by qRT-PCR.MTT assay and Transwell assay were used to detect the changes of proliferation,migration and invasion of Hep-2 cells after inhibiting the expression of SNHG16.Bioinformatics was used to predict target miRNAs for SNHG16,and dual luciferase reporter experiments validated the targeting of SNHG16 and miR-7-5p.Inhibition of miR-7-5p expression verified the effect of SNHG16 on proliferation,migration and invasion of Hep-2 cells.Results The expression level of SNHG16 in la⁃ryngeal carcinoma was significantly increased(P<0.05),and the expression level of miR-7-5p was significantly decreased(P<0.05).Compared with the si-NC group,the inhibition of SNHG16 expression in the si-SNHG16 group significantly inhibited the proliferation of Hep-2 cells[48 h:(0.95±0.09)vs.(0.47±0.05),P<0.05],and the number of migration[(124±12)vs.(51±5),P<0.05]and invasion cells decreased significantly[(115±11)vs.(43±4),P<0.05].SNHG16 could negatively regulate the expression of miR-7-5p.Inhibition of miR-7-5p reversed the inhibitory effect of SNHG16 on proliferation,migration and invasion of Hep-2 cells.Conclusion Inhibition of SNHG16 expression negatively regulates the expression of miR-7-5p and attenuates the proliferation,migration and invasion ability of laryngeal carcinoma cells.