miR-30d在舌鳞状细胞癌组织中表达及反义miR-30d对细胞增殖、侵袭能力的影响

Expression of miR-30d in Tongue Squamous Cell Carcinoma and Effect of Antisense miR-30d on Cell Proliferation and Invasion

目的:探讨miR-30d在舌鳞状细胞癌组织中表达及反义miR-30d对细胞增殖、侵袭能力的影响。方法:选取2013年3月~2015年4月在我院行根治性手术治疗的TSCC患者68例,实时荧光定量PCR检测TSCC组织和癌旁正常组织中miR-30d基因表达,培养舌癌SCC-4细胞,分为ASO-miR-30d组、ASO-阴性对照组和空白对照组,检测各组细胞中miR-30d基因表达,MTT法检测各组细胞增殖能力,Transwell法检测各组细胞迁移和侵袭能力。结果:TSCC组织中miR-30d相对表达量(1.89±0.21),显著高于癌旁正常组织的(1.31±0.16),差异有统计学意义(P=0.000);TSCC组织中miR-30d相对表达量与TNM分期、分化程度和颈淋巴结转移有关(P<0.05);与空白对照组和ASO-阴性对照组比较,ASO-miR-30d组细胞中miR-30d相对表达量降低,差异有统计学意义(P<0.05);MTT实验结果显示,与空白对照组和ASO-阴性对照组比较,ASO-miR-30d组24h、48h、72h和96h时细胞A值均降低,差异均有统计学意义(P<0.05);与空白对照组和ASO-阴性对照组比较,ASO-miR-30d组迁移细胞数和侵袭细胞数均减少,差异均有统计学意义(P<0.05)。结论:miR-30d在TSCC组织中呈高表达,反义miR-30d可有效减少TSCC细胞增殖,抑制细胞迁移和侵袭能力。

Objective.. To investigate the expression of miR--30d in tongue squamous cell carcinoma （TSCC） and effect of antisense miR--30d on cell proliferation and invasion. Methods.. Sixty--eight cases of patients with TSCC underwent radical surgery were selected from March 2013 to April 2015. The expressions of miR--30d in TSCC tis- sues and adjacent normal tissues were detected by using real--time fluorescent quantitative PCR. SCC--4 ceils were cultured and were divided into ASO miR 30d group, ASO negative control group, and blank control group. The expression of miR 30d gene in each group was detected. The cell proliferation in each group was detected by MTT assay. The migration and invasion of cells in each group were detected by Transwell method. Results： The relative expression level of miR--30d in TSCC tissues was 1.89±0.21, which was higher than the adjacent normal tissues （1.31±0. 16）, and the difference was statistically significant （P = 0. 000）. The relative expression level of miR--30d in TSCC tissues was correlated with TNM stage, differentiation, and cervical lymph node metastasis （P 〈0.05）. Compared with the blank control group and the ASO--negative control group, the relative expression lev- el of miR--30d in ASO--miR--30d group was significantly decreased （P〈0.05）. The results of MTT showed that the absorbance A values at 24 h, 48 h, 72 h, and 96 h in ASO--miR--30d group were significantly decreased （P〈0.05）. Compared with the blank control group and the ASO --negative control group, the number of migrating cells and the number of invasive cells in ASO--miR--30d group were decreased （P〈0.05）. Conclusion： miR--30d was highly ex- pressed in TSCC tissues. Antisense miR-- 30d could effectively reduce TSCC cell proliferation and inhibit cell migra- tion and invasion.