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小干扰RNA介导的Snail1基因沉默对转移性乳腺癌细胞系的影响

The Effect of Snail1 Gene Silencing by siRNA in Metastatic Breast Cancer Cell Lines.

作者信息

Aletaha Mansoor, Mansoori Behzad, Mohammadi Ali, Fazeli Mehdi, Baradaran Behzad

机构信息

Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.

Dept. of Pathobiology, Shiraz University, Shiraz, Iran.

出版信息

Iran J Public Health. 2017 May;46(5):659-670.

Abstract

BACKGROUND

Breast cancer is the most common diagnosed cancer among women in the world. Snail1 plays a role in the development of the invasive phenotypes of cancer, neural cell differentiation, cell division and apoptosis in tumor cells. Traces of snail1 in metastasis of breast cancer to bone are observed. The aim of this study was to investigate the effect of specific snail1 siRNAs on the proliferation, migration, induction of apoptosis and cell cycle arrest of MDA-MB-468 cells.

METHODS

In 2015, this experimental study was performed on the MDA-MB-468 cell lines in Immunology Research Center, Tabriz University of Medical Sciences. After the design and construction of siRNA, transfection was performed with transfection reagent. The expression levels of mRNA and protein were measured by qRT-PCR and western blot analysis, respectively. The survival of cells was determined by using MTT assay cells, apoptosis using Tunel assay, Cell migration using scratch assay, Cell cycle analysis by Propidium Iodide (PI) DNA staining method using flow cytometry on the MDA-MB-468.

RESULTS

Transfection with siRNA significantly suppressed the expression of snail1 gene in dose-dependent manner after 48 h (<0.0001). Surprisingly, treatment with snail1 siRNA arrested cell cycle in S phases (<0.0001). Moreover, siRNA transfection had effects on breast adenocarcinoma cells and inhibited the migration (<0.0001), proliferation (<0.0001) and induced apoptosis (<0.0016).

CONCLUSION

The snail1 can be considered as a potent adjuvant in breast cancer therapy.

摘要

背景

乳腺癌是全球女性中最常被诊断出的癌症。Snail1在癌症侵袭性表型的发展、神经细胞分化、肿瘤细胞的细胞分裂和凋亡中发挥作用。在乳腺癌向骨转移中观察到了Snail1的踪迹。本研究的目的是探讨特异性Snail1小干扰RNA(siRNAs)对MDA-MB-468细胞增殖、迁移、凋亡诱导和细胞周期阻滞的影响。

方法

2015年,在大不里士医科大学免疫学研究中心对MDA-MB-468细胞系进行了本实验研究。在设计和构建siRNA后,使用转染试剂进行转染。分别通过qRT-PCR和蛋白质印迹分析测量mRNA和蛋白质的表达水平。使用MTT法测定细胞存活率,使用Tunel法测定细胞凋亡,使用划痕试验测定细胞迁移,使用碘化丙啶(PI)DNA染色法通过流式细胞术对MDA-MB-468细胞进行细胞周期分析。

结果

转染siRNA后48小时,以剂量依赖方式显著抑制了Snail1基因的表达(<0.0001)。令人惊讶的是,用Snail1 siRNA处理使细胞周期停滞在S期(<0.0001)。此外,siRNA转染对乳腺腺癌细胞有影响,抑制了迁移(<0.0001)、增殖(<0.0001)并诱导了凋亡(<0.0016)。

结论

Snail1可被视为乳腺癌治疗中的一种有效佐剂。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/20a6/5442279/e6b2ffc72a6a/IJPH-46-659-g001.jpg

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