长链非编码RNA MT1JP通过FBXW7调控口腔鳞癌细胞生物学活性

Long non-coding RNA-MT1JP regulates the biological activity of oral squamous cell carcinoma cells via FBXW7

目的:探讨长链非编码RNA-MT1JP(long non-coding RNA-MT1JP,LncRNA-MT1JP)通过FBXW7对口腔鳞癌细胞增殖、凋亡、迁移、侵袭等生物学活性的影响。方法:qRT-PCR检测LncRNA-MT1JP在Cal-27、SCC-4、TSCCA口腔鳞癌细胞系和正常口腔上皮细胞系HOK中的表达,将TSCCA细胞系分成LncRNA-MT1JP基因沉默组和对照组,随后采用Lipofectamine TM 2000分别转染LncRNA-MT1JP-shRNA及对照序列LncRNA-MT1JP-NC;MTT法检测两组TSCCA细胞增殖;Transwell小室实验检测两组TSCCA细胞系迁移、侵袭能力;流式细胞仪检测两组TSCCA细胞系凋亡能力;Western blot法检测FBXW7基因蛋白的表达。结果:与正常口腔上皮细胞系HOK比较,LncRNA-MT1JP的相对表达量在口腔鳞癌细胞系Cal-27、SCC-4及TSCCA中显著增高(P<0.01)。TSCCA细胞系在转染24及48 h后,MT1JP基因沉默组与对照组比较OD值无显著的统计学差异(P>0.05);但在72及96 h后OD值显著降低(P<0.05)。与对照组比较,MT1JP基因沉默组TSCCA细胞迁移与侵袭数目显著降低(P<0.01)、凋亡率以及FBXW7基因蛋白相对表达量均显著升高(P<0.05)。结论:LncRNA-MT1JP在口腔鳞癌细胞系呈高表达,MT1JP基因沉默有效抑制了TSCCA细胞系的增殖、迁移、侵袭,促进其凋亡,该作用可能与MT1JP基因沉默后上调抑癌基因FBXW7的表达有关。

Objective:To investigate the long non-coding RNA-MT1JP(LncRNA-MT1JP)to regulate the biological activity of oral squamous cell carcinoma cells by FBXW7.Methods:qRT-PCR was used to detect the expression of LncRNA-MT1JP in Cal-27,SCC-4,TSCCA oral squamous cell carcinoma cell line and normal oral epithelial cell line HOK.The TSCCA cell line was divided into LncRNA-MT1JP gene silencing group and control group.Subsequently,Lipfetamine TM 2000 was used to transfect LncRNA-MT1JP-shRNA and control sequence LncRNA-MT1JP-NC respectively.MTT assay was used to detect the proliferation of TSCCA cells.Transwell chamber assay was used to detect the migration and invasion ability of TSCCA cell lines.The ability of TSCCA cell line to apoptosis was detected by flow cytometry.The expression of FBXW7 gene protein was detected by Western blot.Results:Compared with the normal oral epithelial cell line HOK,the relative expression of LncRNA-MT1JP was significantly increased in oral squamous cell carcinoma cells Cal-27,SCC-4 and TSCCA(P<0.01).There was no statistically significant difference in OD between the MT1JP gene silencing group and the control group after transfection for 24 and 48 h(P>0.05),but the OD value was significantly decreased after 72 and 96 h(P<0.05).Compared with the control group,the number of migration and invasion of TSCCA cells in the MT1JP gene silencing group was significantly decreased(P<0.01).The apoptotic rate and the relative expression of FBXW7 gene protein were significantly increased(P<0.05).Conclusion:LncRNA-MT1JP is highly expressed in oral squamous cell carcinoma cell line.MT1JP gene silencing effectively inhibits the proliferation,migration and invasion of TSCCA cell line and promotes its apoptosis.This effect may be related to up-regulation of tumor suppressor gene FBXW7 after silencing of MT1JP gene.