华西口腔医学杂志2017，Vol.35Issue(6)：576-582,7.DOI:10.7518/hxkq.2017.06.003
体外沉默二牛龙牛儿基转移酶Ⅰ对舌癌细胞迁移和侵袭的影响
Effects of geranylgeranyltransferaseⅠ gene silencing by RNA interference on the migration and invasion of tongue carcinoma
摘要
Abstract
Objective RNA interference was used to silence geranylgeranyltransferaseⅠ(GGTase-Ⅰ)in vitro and to study the effect of GGTase-Ⅰ on the migration and invasion of tongue squamous cancer cells.Methods Three small interfering RNAs (siRNA) were designed according to the GGTase-Ⅰ sequence by Genebank and were transfected into tongue squamous cancer cells Cal-27 to knock down GGTase-Ⅰ expression. The tested cells were divided into three groups, as follows: the RNA-interfered groups (GGTase-Ⅰ siRNA1,GGTase-Ⅰ siRNA 2, GGTase-Ⅰ siRNA 3), a negative control group (disrupted by random sequence NC-siRNA), and a blank control group. GGTase-Ⅰ and RhoA gene expressions were examined by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. The optimum interference group was screened by qRT-PCR and Western blot and was assigned as the experimental group. Matrix metalloproteinase (MMP)-2 and MMP-9 protein expressions were examined by Western blot. GTP-RhoA expression of protein was examined by GST-pull down. The migration and invasion abilities were analyzed by wound healing assay and Transwell motility assay.Results GGTase-Ⅰ mRNA and protein expression in Cal-27 decreased significantly after transfection of GGTase-I siRNA (P<0.05). No significant difference of RhoA gene expression was detected. MMP-2, MMP-9, and GTP-RhoA protein expressions decreased significantly (P<0.05). The migration and invasion abilities were inhibited (P<0.05).Conclusion To inhibit GGTase-Ⅰ expression, the migration and invasion abilities of tongue squa-mous cancer cells should also be inhibited. Further studies on GGTase-Ⅰ may provide novel effective molecular targets for tongue squamous cancer cells.关键词
二牛龙牛儿基转移酶Ⅰ/RhoA/舌癌/迁移/侵袭/基质金属蛋白酶-2/基质金属蛋白酶-9Key words
geranylgeranyltransferaseⅠ/RhoA/tongue squamous cancer/migration/invasion/matrix metallo-proteinase-2/matrix metalloproteinase-9分类
医药卫生引用本文复制引用
陈正岗,郭大伟,王莹,王树人,李敬华,韩金宏,王奇民,童磊,刘文君,杨芳,郭庆圆..体外沉默二牛龙牛儿基转移酶Ⅰ对舌癌细胞迁移和侵袭的影响[J].华西口腔医学杂志,2017,35(6):576-582,7.基金项目
National Natural Science Foundation of China (81372908) （81372908）
Qingdao Municipal Commission of Health and Family Planning (2014-WJZD009, 2013-WSZD011). 国家自然科学基金面上项目(81372908) （2014-WJZD009, 2013-WSZD011）
青岛市卫计委计划项目(2014-WJZD009,2013-WSZD011) （2014-WJZD009,2013-WSZD011）
