LUCAT1/微小RNA-181a-5p对喉癌细胞增殖和化疗敏感度的影响

Effect of LUCAT1/microRNA-181a-5p on proliferation and chemotherapy sensitivity of laryngeal cancer cells

目的探讨LUCAT1/微小RNA(miR)-181a-5p对喉癌细胞增殖和化疗耐药的影响及其机制。方法选取2017年1月至2022年1月我院手术治疗的65例喉癌组织和癌旁组织作为研究对象。采用荧光定量聚合酶链反应(PCR)分析两组组织LUCAT1和miR-181a-5p的表达水平;采用生物信息学和双荧光素酶报告基因分析LUCAT1和miR-181a-5p的关系。人喉癌细胞系Hep-2采用顺铂诱导传声顺铂耐药喉癌细胞系(Hep-2/R)。采用LUCAT1短发卡RNA(shRNA)和对照长链非编码RNA(lncRNA)慢病毒感染Hep-2/R,分别为LUCAT1KD组和对照组。采用细胞计数试剂盒(CCK-8)和体外移植瘤实验分析两组细胞增殖和耐药的变化;采用流式细胞术分析顺铂对两组细胞凋亡的影响。组间计量资料比较采用t检验。结果耐药喉癌组织中LUCAT1表达水平(2.19±0.32)明显高于非耐药喉癌组织(1.61±0.20),差异有统计学意义(t=8.967,P<0.05)。耐药喉癌组织中miR-181a-5p表达水平(1.10±0.13)明显低于非耐药喉癌组织(1.56±0.22),差异有统计学意义(t=9.907,P<0.05)。LUCAT1KD组细胞吸光度(A)值(1.54±0.09)明显低于对照组(1.96±0.06),差异有统计学意义(t=9.164,P<0.05)。经顺铂治疗后,LUCAT1KD组细胞成瘤体积[(486.33±33.84)mm^(3)]明显低于对照组[(732.33±79.13)mm^(3)],差异有统计学意义(t=7.001,P<0.05)。LUCAT1KD组细胞成瘤质量[(4.43±0.64)g]明显低于对照组[(6.18±0.43)g],差异有统计学意义(t=5.575,P<0.05)。经顺铂处理后,LUCAT1KD组细胞凋亡率[(27.09±4.18)%]明显低于对照组[(9.03±1.68)%],差异有统计学意义(t=9.812,P<0.05)。LUCAT1 KD组细胞miR-181a-5p表达水平(1.46±0.16)明显高于对照组(1.06±0.10),差异统计学意义(t=5.090,P<0.05)。结论LUCAT1在喉癌组织中高表达,通过吸附结合miR-181a-5p,调控喉癌细胞顺铂耐药敏感性。

Objective To investigate the effect of LUCAT1/microRNA(miR)-181a-5p on the pro-liferation and chemotherapy resistance of laryngeal cancer cells and its molecular mechanism.Methods A total of 65 cases of laryngeal cancer tissues and adjacent tissues treated surgically in our hospital from January 2017 to January 2022 were selected as the research subjects.The expression levels of LUCAT1 and miR-18la-5p were detected by fluorescence quantitative polymerase chain reaction(PCR).The relation-ship between LUCAT1 and miR-18la-5p was analyzed by bioinformatics and double Luciferase reporter genes.The human laryngeal cancer cell line Hep-2 was induced by cisplatin to establish cisplatin resistant laryngeal cancer cell lines(Hep-2/R).Hep-2/R cells were infected with LUCAT1 short hairpin RNA(shRNA)and control long non-coding RNA(IncRNA)Lentivirus as LUCAT1 KD group and control group,respectively.The cell proliferation and drug resistance were analyzed by cell counting kit-8(CCK-8)assay and in vitro tumor transplantation experiments.The apoptosis rate was analyzed by flow cytometry.The comparison of measurement data between groups was conducted using t-test.Results The expression level of LUCAT1 in drug resistant laryngeal cancer tissue(2.19±0.32)was significantly higher than that in non-drug resistant laryngeal cancer tissue(1.61±0.20,t=8.967,P<0.05).The expression level of miR-18la-5p in drug-resistant laryngeal cancer tissue(1.10±0.13)was significantly lower than that in non-drug-resistant laryngeal cancer tissue(1.56±0.22,t=9.907,P<0.05).The absorbance(A)value of LUCATI KD group(1.54±0.09)was significantly lower than that of the control group(1.96±0.06,t=9.164,P<0.05).After treatment with cisplatin,the tumor volume of LUCAT1 KD group[(486.33±33.84)mm^(3)]was significantly smaller than that of the control group[(732.33±79.13)mm^(3),t=7.001,P<0.05].The tumor mass of LUCAT1 KD group[(4.43±0.64)g]was significantly lower than that of the control group[(6.18±0.43)g,t=5.575,P<0.05].After treatment with cisplatin,the apoptosis rate of LUCATi KD group[(27.09±4.18)%]was significantly lower than that of the control group[(9.03±1.68)%,t=9.812,P<0.05].The expression level of miR-181a-5p in LUCAT1 KD group(1.46±0.16)was significantly higher than that in the control group(1.06±0.10,t=5.090,P<0.05).Conclusion LUCATI is highly expressed in laryngeal cancer tissue and regulates the sensitivity of laryngeal cancer cells to cisplatin resistance through adsorption binding to miR-18la-5p.