基因沉默
小干扰RNA
细胞周期
乳腺癌
细胞生长
活力测定
细胞
癌症研究
分子生物学
免疫印迹
癌症
细胞培养
MTT法
生物
化学
转染
基因
生物化学
遗传学
作者
Hong Peng,Yi Luo,Ziwei Wang,Lin Zeng
出处
期刊:Chin J Endocr Surg
日期:2018-04-25
卷期号:12 (2): 109-114
标识
DOI:10.3760/cma.j.issn.1674-6090.2018.02.006
摘要
Objective
To study the effect of CDCA5 silencing on the growth of human breast cancer cell line MDA-MB-435s in vitro and its mechanism.
Methods
The expression of CDCA5 in breast cancer tissue, the adjacent normal tissue and human breast cancer cell lines MCF-7, MDA-MB-231 and MDA-MB-435s was tested by Western blot. The target sequences of CDCA5 small interfering RNA (siRNA) , control siRNA (siRNA-NC) and blank groups were designed. PCR was used to observe the gene silencing effect on CDCA5. The cell viability was measured by MTT assay in siRNA, siRNA-NC and Blank groups. Cell cycle was determined by flow cytometry. Plk1, SA2 and pSA2 protein expression was tested by Western blot.
Rssults
The expression of CDCA5 was higher in breast cancer tissue than in the adjacent normal tissue. The expression of CDCA5 was higher in human breast cancer cell line MDA-MB-435s than in MCF-7 or MDA-MB-231 cell line. The expression level of CDCA5 was downregulated significantly in siRNA group than other groups. The cell viability was decreased and the cell percentage of G2/M phase was elevated in siRNA group compared with those in siRNA-NC and Blank groups. Inhibition of the CDCA5 gene can suppress the expression of Plk1 and pSA2.
Conclusion
Silencing CDCA5 expression can effectively inhibit the proliferation of breast cancer cells, whose mechanism may lie in arresting cell cycle in G2/M phase by suppressing CDCA5/Plk1/SA2 signaling pathway.
Key words:
Breast cancer; CDCA5; Small interfering RNA
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