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目的筛选成功转染Id1 miRNA真核表达载体乳腺癌MDA-MB-23 1细胞系,观察对乳腺癌细胞的对Id 1、MMP 9表达的影响。方法构建Id1miRNA真核表达载体,转染乳腺癌MDA-MB-231细胞株以杀稻瘟霉素筛选稳定表达的细胞株。经RT-PCR方法鉴定其对Id1、MMP9表达的影响。结果经过筛选,获得转染Id1 miRNA真核表达载体稳定表达的MDA-MB-231细胞系。经RT-PCR方法鉴定,结果表明转染Id1 miRNA真核表达载体的乳腺癌MDA-MB-231细胞株其Id 1、MMP 9表达水平低于对照组,且Id 1、MMP 9表达水平正相关。未转染组与阴性对照转染组无明显差异。结论本实验成功筛选出稳定表达Id1 miRNA的乳腺癌细胞株,证实Id1 miRNA真核表达载体构建成功干扰有效,并且Id1与mmp9表达相关,为将来进一步研究他们之间的生物学行为差异,或详细了解其分子作用机制奠定了基础。
OBJECTIVE: To screen the breast cancer cell line MDA-MB-23 1 successfully transfected with Id1 miRNA and investigate the effect on the expression of Id 1 and MMP 9 in breast cancer cells. Methods The eukaryotic expression vector of Id1miRNA was constructed and transfected into breast cancer cell line MDA-MB-231 to screen the stable cell line with blastomycin. The effect of Id1 and MMP9 on the expression of Id1 and MMP9 was identified by RT-PCR. Results After screening, the MDA-MB-231 cell line transfected with Id1 miRNA eukaryotic expression vector was obtained. The results of RT-PCR showed that the expression of Id 1 and MMP 9 in breast cancer cell line MDA-MB-231 transfected with Id1 miRNA eukaryotic expression vector was lower than that in control group, and the expression of Id 1 and MMP 9 was positively correlated . There was no significant difference between untransfected group and negative control group. Conclusion The experiment successfully screened Id1 miRNA stable expression of breast cancer cell lines confirmed that Id1 miRNA eukaryotic expression vector constructed successfully interfere with effective, and Id1 and mmp9 expression, for further study of their biological behavior differences between, or in detail Understand the molecular mechanism of action laid the foundation.