长链非编码RNA HOXA远端转录本在口腔鳞状细胞癌中的表达及生物学意义

Expression and biological significance of long non-coding RNA HOXA transcript at the distal tip in oral squamous cell carcinoma

目的:探讨长链非编码RNA HOXA远端转录本(HOXA transcript at the distal tip,HOTTIP)在口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)中的表达及对OSCC细胞生物学功能的影响。方法:收集2020年1月至2021年6月在郑州大学第一附属医院接受手术治疗的52例OSCC患者,采用RT-qPCR检测HOTTIP在OSCC组织和癌旁组织以及人OSCC细胞株HSC-4、SCC-9和CAL-27和人正常口腔角质细胞株HOK中的表达水平,并分析其与OSCC患者临床病理资料及总生存期(overall survival,OS)的关系。采用CCK-8、流式细胞术、Transwell和Western blot分析下调HOTTIP或共下调HOTTIP和miR-637对CAL-27细胞增殖、凋亡、迁移、侵袭和上皮间质转化(epithelial-mesenchymal transition,EMT)能力的影响。采用双荧光素酶报告基因实验分析HOTTIP和miR-637的靶向关系。结果:与癌旁组织相比,OSCC组织中HOTTIP的表达水平明显升高(P<0.05),且与肿瘤大小、TNM分期、分化程度、淋巴结转移及OS相关(P<0.05)。与人正常口腔角质HOK细胞相比,HSC-4、SCC-9和CAL-27细胞中HOTTIP的表达明显升高(P<0.05)。下调HOTTIP后,CAL-27细胞的增殖、迁移、侵袭及EMT能力明显减弱,细胞凋亡率明显升高(P<0.05)。HOTTIP与miR-637具有靶向关系,且下调HOTTIP后,CAL-27细胞中miR-637表达明显升高(P<0.05)。同时下调HOTTIP和miR-637后,CAL-27细胞的增殖、迁移、侵袭与EMT能力明显增强,细胞凋亡率明显降低(P<0.05)。结论:HOTTIP在OSCC组织和细胞中高表达,可靶向miR-637调控OSCC细胞的增殖、凋亡、迁移、侵袭和EMT能力。

Objective:To investigate the expression of long non-coding RNA HOXA transcript at the distal tip(HOTTIP)in oral squamous cell carcinoma(OSCC)and examine its effect on the biological function of OSCC cells.Methods:A total of 52 patients with OSCC who received surgical treatment in The First Affiliated Hospital of Zhengzhou University from January 2020 to June 2021 were collected.The expression levels of HOTTIP in OSCC and adjacent tissues,human OSCC cell lines HSC-4,SCC-9 and CAL-27,and human normal oral keratinocyte line HOK were detected by quantitative real-time polymerase chain reaction(RT-qPCR),and its correlation with clinicopathological data and overall survival(OS)of OSCC patients was analyzed.The effects of down-regulating HOTTIP or co-down-regulating HOTTIP and miR-637 on the proliferation,apoptosis,migration,invasion,and epithelial-mesenchymal transition(EMT)of CAL-27 cells were analyzed using Cell Counting Kit-8(CCK-8),flow cytometry,Transwell assays,and Western blot.The targeting relationship between HOTTIP and miR-637 was analyzed by dual luciferase reporter gene assays.Results:The expression level of HOTTIP in OSCC tissues was significantly increased compared with adjacent tissues(P<0.05)and was correlated with tumor size,TNM stage,degree of differentiation,lymph node metastasis,and OS(P<0.05).The expression of HOTTIP in HSC-4,SCC-9,and CAL-27 cells significantly increased compared with normal HOK cells(P<0.05).After down-regulation of HOTTIP expression,the proliferation,migration,invasion,and EMT abilities of CAL-27 cells were significantly weakened,whereas the apoptosis rate significantly increased(P<0.05).HOTTIP had a targeting relationship with miR-637;after down-regulating HOTTIP,the expression of miR-637 in CAL-27 cells significantly decreased(P<0.05).After the simultaneous downregulation of HOTTIP and miR-637,the proliferation,migration,invasion,and EMT abilities of CAL-27 cells were significantly enhanced,whereas the apoptosis rate significantly decreased(P<0.05).Conclusions:HOTTIP is highly expressed in OSCC tissues and cells and can target miR-637 to regulate the proliferation,apoptosis,migration,invasion,and EMT of OSCC cells.