microRNA沉默SMYD3基因对乳腺癌细胞Wnt/β-Catenin通路及c-Myc基因影响的研究

Effects of Silencing SMYD3 Gene on Wnt/β-catenin Pathway and c-Myc Gene in Breast Cancer Cells in vitro

目的研究沉默SMYD3基因后,乳腺癌细胞MDA-MB-231中Wnt/β-catenin通路和c-Myc基因的变化,探讨其可能机制。方法构建携带绿色荧光蛋白基因的SMYD3-microRNA真核表达质粒载体,脂质体法稳定转染MDA-MB-231细胞,实时定量PCR检测SMYD3、Wnt10b、β-catenin、c-Myc mRNA的表达。结果流式细胞检测转染效率达到95%以上,实验组SMYD3、Wnt10b、c-Myc mRNA水平低于空白对照组(P<0.05),β-catenin水平高于空白对照组(P<0.05);阴性对照组与空白对照组相比无明显变化。结论沉默SMYD3基因可直接或通过Wnt/β-catenin通路下调c-Myc基因的表达,从而减弱肿瘤细胞的生长及侵袭转移能力,为乳腺癌的临床治疗提供了新的基因靶点。

Objective To observe the alteration of Wnt/β - catenin pathway and c - Myc gene after silencing SMYD3 gene in breast cancer cells MDA - MB - 231 and explore the possible mechanism. Methods Eukaryotie expression plasmids of mieroRNA targeting SMYD3, which carried the green fluorescent protein gene, were constructed and stably transfacted into MDA - MB -231 cells. The expression levels of SMYD3, Wnt10b,β - catenin, e - Mye mRNA were detected by realtime PCR. Results The transfection efficiency detected by Flow cytometrie was up to above 95%. SMYD3, Wnt10b, e - Myc mRNA expression levels in the group with silencing SMYD3 gene were lower than those in the control group （ P 〈 0.05 ） , while β - eatenin mRNA expression level in the group with silencing SMYD3 gene was higher than that in the control group（P 〈 0.05）. Conclusion Silencing SMYD3 gene can be directly or through Wnt/β - catenin pathway down - regulate e - Myc gene expression, thus weakening the growth and invasion of breast cancer cell. We will have a new target gene for the clinical therapy of breast cancer.