摘要
背景:研究发现肿瘤相关基因以及肿瘤基因通路的改变在肿瘤的形成与发展中起重要作用。目的:探讨siRNA干扰技术沉默FABP-5基因表达对人宫颈癌SiHa细胞生物学特性的影响。方法:以人宫颈癌SiHa细胞为研究对象,根据FABP-5 mRNA编码序列设计并合成干扰si RNA序列,瞬时转染SiHa细胞。用RT-PCR和Western blot法分别从mRNA水平和蛋白水平检测FABP-5基因的表达;流式细胞技术检测各组细胞周期的变化;CCK-8法测定细胞体外增殖能力;细胞侵袭小室法检测各组细胞的体外侵袭力;TUNEL染色检测各组细胞的凋亡情况。结果与结论:与转染空病毒阴性对照组和未转染空白对照组比较,FABP-5 siRNA组FABP-5 mRNA和蛋白表达明显下降,细胞生长速度明显减慢,细胞周期阻滞在G_0/G_1期,S期细胞数减少,细胞侵袭力显著下降,细胞凋亡率明显升高。结果说明siRNA技术可以成功诱导FABP-5基因沉默,从而影响宫颈癌SiHa细胞的的生长、增殖和凋亡。
BACKGROUND： Cancer related genes and pathways play a critical role in formation and development of cancer. OBJECTIVE： To investigate the silencing epidermal fatty acid binding protein 5（FABP-5） by siRNA interference on biological characteristics of the cervical carcinoma cell line SiHa. METHODS： Design and synthesis of siRNA interference sequence used to transiently transfect SiHa cells was performed according to FABP-5 mRNA coding sequence. mRNA and protein expressions of FABP-5 were detected by RT-PCR and western blot assay, respectively. Meanwhile, cell cycle, proliferation, invasion and apoptosis were detected by flow cytometry, cell counting bit-8 assay, Boyden chamber assay, and TUNEL staining, respectively. RESULTS AND CONCLUSION： FABP-5 mRNA and protein expressions, cell growth speed, cell number at S phase（cell cycle was arrested at the G_0/G_1 phase） and cell invasion were significantly decreased, while the cell apoptosis was significantly increased in FABP-5 siRNA group compared with the negative control and blank control groups. Our findings indicate that the specific silencing FABP-5 gene expression by siRNA interference can inhibit SiHa cervical cell growth and proliferation, but accelerate cell apoptosis.
出处
《中国组织工程研究》
CAS
北大核心
2016年第15期2218-2224,共7页
Chinese Journal of Tissue Engineering Research
