摘要
目的探讨沉默癌基因c-myc对人成釉细胞瘤细胞株hTERT^+-AM的基质金属蛋白酶(MMP)表达及细胞侵袭力的影响。方法利用慢病毒载体Lenti-shc-myc-eGFP/puro稳定转染hTERT^+-AM细胞,成功构建c-myc沉默稳转细胞株。利用定量反转录聚合酶链反应(RT-PCR)检测细胞中c-myc、MMP-2、MMP-9的mRNA表达情况。蛋白印迹法(Western blot)检测细胞中c-myc、MMP-2、MMP-9的蛋白表达情况,利用Transwell实验检测其迁移侵袭能力的变化。所有数据采用SPSS 13.0统计软件包进行统计学分析。结果沉默c-myc基因后hTERT^+-AM细胞中c-myc的mRNA相对表达量为0.41±0.02,下降约60%(LSD-t=-0.591,P<0.001),而其蛋白的表达也下调约50%(LSD-t=-0.461,P<0.001),MMP-2、MMP-9的mRNA相对表达量分别为0.79±0.05和0.76±0.01,均下降约20%(LSD-t_(MMP-2)=-0.206,LSD-t_(MMP-9)=-0.242,P<0.001),而MMP-2、MMP-9的蛋白表达均约下调了50%(LSD-t_(MMP-2)=-0.257,LSD-t_(MMP-9)=-0.261,P<0.001),细胞迁移(±s=24.9±4.5,LSD-t=-17.000,P=0.021)、侵袭(±s=10.8±4.1,LSD-t=-22.250,P=0.011)能力均下降。结论 c-myc基因参与人成釉细胞瘤AM局部侵袭的调节,沉默c-myc基因后,继而下调MMP-2、MMP-9,抑制AM细胞的迁移和侵袭能力,c-myc可能成为人成釉细胞瘤AM侵袭防治的新靶点。
Objective To investigate the effect of c-myc silence on MMPs expression and invasion of immortalized ameloblastoma cell line hTERT^＋-AM. Methods Functional characteristics of immortalized ameloblastoma cell line hTERT^＋-AM with stable c- myc silence included migration, invasion, and regulation of matrix metalloproteinases MMP-2, MMP-9 by qRT-PCR and Western blotting. All data were analyzed by SPSS 13.0 software package. Results After gene silencing of c-myc in hTERT＋-AM cells, the mRNA relative expression of c-myc, MMP-2 and MMP-9 was （0.41 ± 0.02）, （0.79 ± 0.05） and （0.76 ± 0.01 ） respectively, declined by about 60% （LSD-t =-0.591, P 〈 0.001 ） and 20% （LSD-tMMp.2 =-0.206, LSD-tMMe-9 =-0.242, P 〈 0.001 ）, the protein expression of c-myc, MMP-2 and MMP-9 was reduced by about 50% （LSD-te-mtc=-0.461, LSD-tMMp-2 =-0.257, LSD-tMMP.9 =-0.261, P 〈 0.001） , the capacity of migration （x±s =24.9 ＋ 4.5,LSD-t=-17.0OO,P=O.021 ） and invasion （x±s = 10.8±4.1 ,LSD-t=-22.250,P = 0.011 ） was decreased. Conclusions Silence of c-myc gene significantly inhibits MMPs expression and cell invasive ability of hTERT^＋-AM cells, suggesting c- myc may be a new target for ameloblastoma treatment.
出处
《中华口腔医学研究杂志（电子版）》
CAS
2017年第5期266-272,共7页
Chinese Journal of Stomatological Research(Electronic Edition)
基金
国家自然科学基金(81072229)
广东省自然科学基金(2014A030313025)
