ELF5基因对人卵巢癌细胞侵袭转移和大鼠移植瘤生长的抑制作用及其机制

发布时间：2018-03-16 08:58

本文选题：卵巢癌　切入点：ELF基因　出处：《山东医药》2017年13期 　论文类型：期刊论文

【摘要】：目的探讨ELF5基因对人卵巢癌细胞侵袭转移和大鼠移植瘤生长的抑制作用以及可能的机制。方法体外实验:人卵巢癌细胞随机分为重组质粒组、空质粒组、未经转染组,重组质粒组、空质粒组分别转染构建好的pcDNA3.1-ELF5+EGFP真核表达载体、空载质粒pcDNA3.1-EGFP,未经转染组未经任何转染。Q-PCR法检测各组ELF5 mRNA的相对表达量,Western blotting法检测基质金属蛋白酶MMP-2及MMP-9蛋白的相对表达量;Traswell试验评价ELF5对卵巢癌细胞侵袭转移的影响。体内试验:建立人卵巢癌NOD/SCID鼠左前肢腋下移植瘤模型,应用随机数字表法将大鼠分为3组,即重组质粒大鼠组(转染pcDNA3.1-ELF5+EGFP的人卵巢癌细胞)、空质粒大鼠组(转染空载质粒的人卵巢癌细胞)、未经转染大鼠组(未经转染的人卵巢癌细胞),将处于对数生长期的3组细胞2×104/m L接种于NOD/SCID小鼠左前肢腋下。对比3组小鼠的移植瘤质量和体积,计算各组小鼠肿瘤抑制率(首先对转移瘤行HE染色进行病理组织学检查以确定其致瘤性)。Western blotting法检测3组NOD/SCID小鼠移植瘤组织中MMP-2及MMP-9蛋白的相对表达量。结果人卵巢癌细胞转染pcDNA3.1-ELF5+EGFP真核表达载体后,可稳定表达,且于72 h后转染率较高;与空质粒组和未经转染组比较,重组质粒组MMP-2及MMP-9蛋白相对表达量降低(P均0.05)。Transell试验中,重组质粒组穿膜细胞数少于空质粒组及未经转染组(P均0.05)。重组质粒大鼠组抑瘤率达64.4%。重组质粒大鼠组与其余两组比较,其瘤组织中MMP-2及MMP-9蛋白相对表达量降低(P0.05),而空质粒大鼠组和未经转染大鼠组比较,P均0.05。结论 ELF5基因可能通过抑制MMP-2及MMP-9的表达而降低卵巢癌细胞的侵袭转移能力。
[Abstract]:Objective to investigate the inhibitory effect of ELF5 gene on invasion and metastasis of human ovarian cancer cells and the growth of transplanted tumor in rats. Methods Human ovarian cancer cells were randomly divided into three groups: recombinant plasmid group, empty plasmid group and untransfected group. The recombinant plasmid group and the empty plasmid group were transfected with the constructed eukaryotic expression vector of pcDNA3.1-ELF5 EGFP, respectively. The relative expression of ELF5 mRNA in the untransfected group was detected without any transfection. The relative expression of matrix metalloproteinase MMP-2 and MMP-9 protein in each group was detected by Western blotting. The invasion and metastasis of ELF5 to ovarian cancer cells were evaluated by Traswell test. Effect. In vivo test: establish the model of left forelimb axillary transplantation tumor in NOD/SCID mice of human ovarian cancer, The rats were divided into three groups by random digital table. The recombinant plasmid group (human ovarian cancer cells transfected with pcDNA3.1-ELF5 EGFP, empty plasmid rats) and untransfected rats (untransfected human ovarian cancer cells) will be in logarithmic growth phase. The 3 groups were inoculated with 2 脳 10 4 / mL cells under the left forelimb axilla of NOD/SCID mice. The tumor mass and volume of the transplanted tumor were compared in the three groups. The tumor inhibition rate of each group was calculated. First, the tumor metastasis was stained by HE staining to determine the relative expression of MMP-2 and MMP-9 protein in the transplanted tumor tissues of NOD/SCID mice. Results the expression of MMP-2 and MMP-9 protein in the transplanted tumor tissues of three groups of NOD/SCID mice was determined by the method of HE staining to determine the relative expression of MMP-2 and MMP-9 protein in the transplanted tumor tissues of the three groups. After transfection of pcDNA3.1-ELF5 EGFP eukaryotic expression vector into Nest cancer cells, The relative expression of MMP-2 and MMP-9 protein in the recombinant plasmid group was lower than that in the blank plasmid group and the untransfected group, and the relative expression of MMP-2 and MMP-9 protein in the recombinant plasmid group was lower than that in the blank plasmid group and the untransfected group. The number of perforated cells in the recombinant plasmid group was less than that in the blank plasmid group and the untransfected group (P < 0.05). The tumor inhibition rate in the recombinant plasmid group was 64.4.The recombinant plasmid group was compared with the other two groups. The relative expression of MMP-2 and MMP-9 protein in tumor tissue decreased P0.05, while that in blank plasmid group was 0.05 compared with that in untransfected group. Conclusion ELF5 gene may reduce the invasion and metastasis ability of ovarian cancer cells by inhibiting the expression of MMP-2 and MMP-9.
【作者单位】：徐州医科大学研究生院;徐州医科大学附属医院;
【分类号】：R737.31

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