靶向抑制MAP3K1对小鼠乳腺癌细胞生物学行为的影响  

Effect of targeted inhibition of MAP3K1 gene on the biological behavior of mouse breast cancer cells

作　　者：王胜洁[1,2] 刘春[2] 汪海峰[1] 吴刘成[2] 彭晓清[2] 邵义祥[2] 

机构地区：[1]南京医科大学康达学院细胞生物学与组织胚胎学教研室,江苏连云港222000 [2]南通大学实验动物中心,江苏南通226001

出　　处：《黑龙江畜牧兽医》2016年第11期26-31,290,共7页Heilongjiang Animal Science And veterinary Medicine

基　　金：江苏省高校自然科学研究项目(14KJB180016);江苏省高校自然科学研究项目(16KJD180005);南京医科大学康达学院科研发展基金项目(KD2015KYJJYB001)

摘　　要：为了在细胞学水平研究靶向抑制MAP3K1(mitogen-activated protein kinase kinase kinase 1)基因对小鼠乳腺癌4T1细胞黏附、运动及侵袭能力等生物学行为的影响,试验用体外构建靶向"沉默"MAP3K1基因的amiRNA(artificial microRNA)干扰载体,以X-treme GENE HP转染4T1细胞(干扰组转染amiRNA干扰载体,对照组转染空载体,空白组未转染质粒)对MAP3K1基因mRNA及蛋白表达水平进行检测,以测定干扰效率;分别以黏附试验检测小鼠乳腺癌4T1细胞的黏附能力,用划痕-愈合试验与Transwell侵袭试验测定小鼠乳腺癌4T1细胞的运动与侵袭能力。结果表明:小鼠乳腺癌4T1细胞转染MAP3K1 amiRNA-3干扰载体与空载体后,干扰4T1细胞MAP3K1 mRNA水平及蛋白水平均被有效抑制,干扰效率高达70%(P<0.01)。试验组4T1细胞黏附能力极显著低于对照组(P<0.01);干扰组4T1细胞的运动与侵袭能力均显著低于对照组(P<0.05)。说明构建的amiRNA干扰载体能够有效抑制靶基因MAP3K1的表达,并能够显著抑制小鼠乳腺癌4T1细胞的黏附、运动与侵袭能力;靶向抑制MAP3K1基因能够广泛地影响小鼠乳腺癌4T1细胞的生物学行为。To study the effect of targeted inhibition of mitogen- activated protein kinase kinase kinase 1 （MAF3K1）gene on the adhesion, mi- gration, invasion abilities, and other biological behaviors of mouse 4T1 breast cancer cells at the cytological level, an artificial microRNA （amiRNA） interference vector targeting silent MAP3K1 gene was constructed in vitroin the test. X - tremeGENE HP was used to transfecttbe 4T1 cells, thereinto,tbe interferencegroup was transfected with amiRNA interference vector, the control group was transfected with empty vector, and the blank group had non -trandected plasmid. TheMAP3K1 gene mRNA and protein expression levels were detected to determine the inter- ference efficiency. A cell adhesion experiment was used to detect the adhesion ability of the 4T1 cells, and the scratch - healing and Transwell experiments were used to determine the migratory and invasive ability of the 4T1 cells. The results showed that after the 4T1 cells were transfect- ed with MAP3K1 amiRNA -3 interference vector and empty vector, the expression levels of MAF3K1 gene mRNA and protein of the 4T1 cells were effectively inhibited, and the interference efficiency was up to 70% （ P 〈 0.01 ）. The adhesion ability of 4T1 cells in the experimental group was extremely significantly lower （ P 〈 0.01 ） than that in the control group. The migratory and invasive abilities of 4T1 cells in the inter- ference group were significantly lower （P 〈 0.05 ） than those in the control group. The results indicate that the constructed MAP3 K1 amiRNA interference vector can effectively inhibit the expression of target gene MAIr3K1, and can significantly inhibitthe adhesion, migration and inva- sion abilities of mouse 4T1 breast cancer cells. The targeted inhibition of MAIr3KI gene can widely influence the biological behaviors of mouse 4T1 breast cancer cells.

关 键 词：RNA干扰 MAP3K1 乳腺癌 4T1细胞 黏附能力 运动与侵袭能力 

分 类 号：S865.13[农业科学—野生动物驯养] Q813[农业科学—畜牧兽医]