LncRNA CASC9通过调控miR-423-5p/NRSN2轴促进口腔鳞状细胞癌细胞增殖并抑制细胞凋亡

LncRNA CASC9 Promotes Proliferation and Inhibits Apoptosis of Oral Squamous Cell Carcinoma Cells by Regulating miR-423-5p/NRSN2 Axis

口腔鳞状细胞癌(OSCC)是一种侵袭性强的头颈部恶性肿瘤,严重威胁着人类的身体健康。该研究旨在探讨长链非编码RNA肿瘤易感候选者9(Lnc RNA CASC9)调控mi R-423-5p/neurensin-2(NRSN2)轴对OSCC细胞增殖和凋亡的影响。采用qRT-PCR检测OSCC癌组织及HSC-3、CAL-27、SCC-15细胞中CASC9水平;将SCC-15细胞分为对照组、sh-NC-1组、sh-CASC9组、miR-NC组、miR-423-5p mimics组、anti-miR-NC组、anti-miR-423-5p组、sh-NC组、sh-NRSN2组、shCASC9+anti-miR-NC组、sh-CASC9+anti-miR-423-5p组,qRT-PCR检测CASC9和miR-423-5p表达,CCK-8、流式细胞术、Western blot分别检测细胞增殖、凋亡及NRSN2蛋白表达情况;裸鼠体内移植瘤实验观察CASC9对肿瘤生长的影响;RNA pull down及双荧光素酶报告基因实验检测miR-423-5p与CASC9、NRSN2的靶向关系。结果显示,在OSCC组织和细胞中CASC9呈高表达,且在SCC-15细胞中CASC9表达量最高,故以SCC-15细胞为研究对象;沉默CASC9可抑制SCC-15细胞增殖、促进凋亡并抑制裸鼠体内肿瘤的形成(P<0.05);CASC9靶向负调控miR-423-5p表达,miR-423-5p靶向负调控NRSN2表达;抑制miR-423-5p可促进SCC-15细胞增殖、抑制凋亡;下调NRSN2可抑制SCC-15细胞增殖、促进凋亡;抑制miR-423-5p表达减弱了沉默CASC9对OSCC细胞所发挥的作用(P<0.05)。总之,沉默CASC9可能通过靶向上调mi R-423-5p来抑制NRSN2表达进而抑制SCC-15细胞的增殖,促进细胞凋亡。

OSCC(oral squamous cell carcinoma)is a highly invasive malignant tumor of the head and neck,which seriously threatens human health.The purpose of the study was to investigate the effects of LncRNA CASC9(long chain non coding RNA tumor susceptible candidate 9)regulating the miR-423-5 p/NRSN2(neuren­sin-2)axis on the proliferation and apoptosis of OSCC cells.qRT-PCR(quantitative real-time PCR)was used to de­tect the expression level of CASC9 in cancer tissues of 21 OSCC patient and hSC-3,CAL-27,SCC-15 cell;divide SCC-15 cells into control group,sh-NC-1 group,sh-CASC9 group,miR-NC group,miR-423-5p mimics group,anti-miR-NC group,anti-miR-423-5p group,sh-NC group,sh-NRSN2 group,sh-CASC9+anti-miR-NC group and sh-CASC9+anti-miR-423-5p group,qRT-PCR was used to detect the expression of CASC9 and miR-423-5p,cell proliferation,apoptosis and the expression of NRSN2 protein were detected by CCK-8,flow cytometry and West­ern blot,respectively;the effect of CASC9 on tumor growth was observed by tumor xenograft experiment in nude mice;RNA pull down and the dual luciferase reporter gene experiment were used to detect the targeting relations hip of CASC9 and miR-423-5p,miR-423-5p and NRSN2.The results show that,CASC9 was highly expressed in OSCC tissues and cells,and the expression of CASC9 was the highest in SCC-15 cells,therefore,SCC-15 cells were used as the research object;silencing CASC9 could inhibit the proliferation of SCC-15,promote apoptosis and inhibit tumor formation in nude mice(P<0.05);CASC9 targeted and negatively regulated the expression of miR-423-5p,and miR-423-5p targeted and negatively regulated the expression of NRSN2;inhibition of miR-423-5p could promote the proliferation and inhibit apoptosis of SCC-15 cells,and down-regulation of NRSN2 could inhibit the proliferation and promote apoptosis of SCC-15 cells;inhibition of miR-423-5p expression weakened the effect of silencing CASC9 on OSCC cells(P<0.05).In short,silencing of CASC9 inhibits the proliferation and promotes apoptosis of SCC-15 cells by targeting up-regulation of miR-423-5p to inhibit the expression of NRSN2.