miR-182-5p调控FHIT对喉鳞癌细胞增殖、凋亡、迁移、侵袭的影响

目的探讨miR-182-5p调控脆性组氨酸三联体(fragile histidine triad,FHIT)对喉鳞癌细胞的增殖、凋亡、迁移、侵袭的影响及其作用机制。方法选取喉鳞癌组织和癌旁正常组织标本各39例,将anti-miR-NC、anti-miR-182-5p、miR-NC、miR-182-5p载体质粒分别转染至FD-LSC-1喉鳞癌细胞中,分别记为anti-miR-NC组、anti-miR-182-5p组、miR-NC组、miR-182-5p组;将anti-miR-182-5p质粒分别与si-NC、si-FHIT载体质粒共转染至FD-LSC-1细胞中,分别记为anti-miR-182-5p+si-NC组、anti-miR-182-5p+si-FHIT组;转染均采用脂质体法。实时荧光定量PCR(RT-qPCR)检测喉癌组织、癌旁组织中的FHIT及miR-182-5p表达水平;蛋白质印迹(Western Blot)法检测FHIT、细胞周期素D1(CyclinD1)、p21、B细胞淋巴瘤/白血病-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、上皮钙黏蛋白(E-cadherin)、基质金属蛋白酶2(MMP-2)的表达;四甲基偶氮唑盐比色法(MTT)检测细胞活性;流式细胞术检测细胞凋亡;Transwell检测细胞迁移和侵袭;双荧光素酶报告基因检测miR-182-5p和FHIT的靶向关系。结果与癌旁组织相比,喉鳞癌组织中miR-182-5p表达水平显著升高,FHIT表达水平显著降低。抑制miR-182-5p表达可降低细胞活性和迁移、侵袭数量,提高细胞凋亡率,降低CyclinD1、Bcl-2、MMP-2表达水平,提高p21、Bax、E-cadherin表达水平。miR-182-5p可靶向调控FHIT,抑制FHIT能逆转抑制miR-182-5p对喉鳞癌细胞FD-LSC-1增殖、迁移、侵袭的抑制和凋亡促进作用。结论抑制miR-182-5p表达可抑制喉鳞癌细胞增殖、迁移和侵袭,促进细胞凋亡,其机制可能与FHIT表达相关。

Effect of miR-182-5p on Proliferation,Apoptosis,Migrationand Invasion of Laryngeal Squamous Carcinoma Cells via FHIT

Objective To investigate the effect of miR-182-5p on proliferation,apoptosis,migration and invasion of laryngeal squamous cell carcinoma cells and its mechanism.Methods The anti-miR-NC,anti-miR-182-5p,miR-NC,and miR-182-5p were transfected into FD-LSC-1 cells,respectively,and recorded as anti-miR-NC group,anti-miR-182-5p group,miR-NC group,miR-182-5p group,respectively.Anti-miR-182-5p plasmid was co-transfected into FD-LSC-1 cells with si-NC and si-FHIT,respectively,and recorded as anti-miR-182-5p+si-NC group,anti-miR-182-5p+si-FHIT group.Transfection was performed by liposome method.Real-time quantitative PCR(RT-qPCR)was used to detect miR-182-5p expression.Western Blot was used to detect expressions of fragile histidine triad(FHIT),Cyclin D1(Cyclin D1),p21,B-cell lymphoma/leukemia-2(Bcl)-2),Bcl-2 related X protein(Bax),epithelial cadherin(E-cadherin),matrix metalloproteinase 2(MMP-2).Tetramethylazozolium salt colorimetric assay(MTT)was used to detect cell viability.Flow cytometry was used to detect apoptosis.Transwell was used to detect cell migration and invasion;dual luciferase reporter gene detects the targeting relationship between miR-182-5p and FHIT.Results Compared with paracancer,the expression of miR-182-5p in laryngeal squamous cell carcinoma was significantly increased,and the expression of FHIT was significantly decreased.Inhibition of miR-182-5p expression reduced cell viability,migration and invasion,increased apoptosis rate,decreased expression of CyclinD1,Bcl-2 and MMP-2,and increased expression of p21,Bax and E-cadherin.miR-182-5p targeted regulation of FHIT,and inhibition of FHIT could reverse the inhibition of miR-182-5p on the proliferation,migration,invasion and apoptosis of FD-LSC-1 cells.Conclusion Inhibition of miR-182-5p expression can inhibit the proliferation,migration and invasion of laryngeal squamous carcinoma cells and promote cell apoptosis.The mechanism may be related to the expression of FHIT.