敲低 FAM64A 抑制乳腺癌细胞的增殖和迁移。

Knockdown of FAM64A suppresses proliferation and migration of breast cancer cells.

机构信息

Department of Occupational Health and Occupational Medicine, School of Public Health, Southern Medical University, Guangzhou, 510515, Guangdong, China.

State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Sun Yat-Sen University Cancer Center, Guangzhou, 510060, Guangdong, China.

出版信息

Breast Cancer. 2019 Nov;26(6):835-845. doi: 10.1007/s12282-019-00991-2. Epub 2019 Jul 1.

DOI:10.1007/s12282-019-00991-2

PMID:31264076

Abstract

BACKGROUND

FAM64A is a mitotic regulator promoting cell metaphase-anaphase transition, and it is frequently reported to be highly expressed in cancer cells. However, the role of FAM64A in human breast cancer (BrC) is poorly studied.

METHODS

The expression of FAM64A mRNA in BrC samples was determined by RT-qPCR assay and TCGA database mining. Kaplan-Meier plotter was used to analyze whether FAM64A expression impacted prognosis. Then, the expression of FAM64A was silenced using RNA interference. Cell-counting assay, colony formation assay and flow cytometry assay were conducted to detect proliferation; transwell migration assay, EMT-related proteins expression (E-cadherin, N-cadherin and vimentin), and EMT-related transcription factors mRNA expression (Snail, Twist, Slug) were conducted to evaluate the migration ability.

RESULTS

FAM64A was highly expressed in human BrC samples, which was negatively associated with poor survival time. Analysis of FAM64A expression in BrC cell lines demonstrated that the expression of FAM64A was significantly correlated with the proliferation rate and migration ability of BrC cells. Indeed, knockdown of FAM64A suppressed the proliferation of MDA-MB-231 and MCF-7 cells. Importantly, we also found that silencing of FAM64A inhibited the migration of BrC cells via impeding epithelial-mesenchymal transition.

CONCLUSIONS

Our findings suggest that FAM64A plays an important role in the proliferation and migration of BrC cells, which might serve as a potential target for BrC treatment.

摘要

背景

FAM64A 是一种有丝分裂调节因子，可促进细胞中期-后期过渡，其在癌细胞中高表达的情况常有报道。然而，FAM64A 在人乳腺癌（BrC）中的作用尚未得到充分研究。

方法

通过 RT-qPCR 检测和 TCGA 数据库挖掘来确定 BrC 样本中 FAM64A mRNA 的表达。Kaplan-Meier plotter 用于分析 FAM64A 表达是否影响预后。然后，采用 RNA 干扰来沉默 FAM64A 的表达。通过细胞计数法、集落形成实验和流式细胞术检测细胞增殖；通过 Transwell 迁移实验、EMT 相关蛋白表达（E-钙黏蛋白、N-钙黏蛋白和波形蛋白）和 EMT 相关转录因子 mRNA 表达（Snail、Twist、Slug）来评估迁移能力。

结果

FAM64A 在人 BrC 样本中高表达，与生存时间较差呈负相关。在 BrC 细胞系中分析 FAM64A 表达情况表明，FAM64A 的表达与 BrC 细胞的增殖率和迁移能力显著相关。事实上，敲低 FAM64A 可抑制 MDA-MB-231 和 MCF-7 细胞的增殖。重要的是，我们还发现，沉默 FAM64A 通过抑制上皮-间充质转化来抑制 BrC 细胞的迁移。