南方医科大学学报

南方医科大学学报 ›› 2023, Vol. 43 ›› Issue (4): 560-567.doi: 10.12122/j.issn.1673-4254.2023.04.08

• • 上一篇    下一篇

抑制RAB27a能够抑制三阴乳腺癌细胞的增殖、侵袭和粘附

王 丽，严志锐，夏耀雄   

  1. 云南省肿瘤医院//昆明医科大学第三附属医院放射治疗科，云南 昆明 650118
  • 出版日期:2023-04-20 发布日期:2023-05-16

Silencing RAB27a inhibits proliferation, invasion and adhesion of triple-negative breast cancer cells

WANG Li, YAN Zhirui, XIA Yaoxiong   

  1. Department of Radiation Oncology, Yunnan Provincial Cancer Hospital (Third Affiliated Hospital of Kunming Medical University), Kunming 650118, China
  • Online:2023-04-20 Published:2023-05-16

RichHTML

20

PDF

161

摘要： 目的 抑制在外泌体分泌过程中发挥核心作用的RAB27蛋白家族的表达，以探究其对三阴性乳腺癌（TNBC）细胞生物学功能的影响。方法 利用荧光定量PCR和蛋白印迹法对比3株三阴性乳腺癌细胞系MDA-MB-231、MDA-MB-468、Hs578T和正常乳腺上皮细胞系MCF10A的RAB27家族基因和蛋白表达以及外泌体的分泌情况；使用siRNA干扰技术分别抑制3株三阴性乳腺癌细胞中RAB27a和RAB27b的表达，并采用荧光定量PCR和蛋白印迹法检测抑制效率，随后使用蛋白印迹法检测这3株三阴性乳腺癌细胞RAB27a和RAB27b分别被抑制后外泌体的分泌情况。用细胞增殖、细胞侵袭和细胞粘附实验评价RAB27a和RAB27b分别被抑制后对这3株三阴性乳腺癌细胞生物学功能造成的影响。结果 三阴性乳腺癌细胞和正常乳腺上皮细胞相比外泌体分泌更加旺盛（P<0.001），并且RAB27a和RAB27b的表达在基因和蛋白水平上均显著上升（P<0.01）。分别抑制3株三阴性乳腺癌细胞RAB27a的表达后，外泌体分泌显著下调（P<0.001），抑制RAB27b的表达对外泌体分泌无显著影响。并且外泌体分泌显著下调后，与外泌体分泌未受到显著影响的三阴性乳腺癌细胞相比，癌细胞增殖能力、细胞侵袭能力和细胞粘附能力均显著下调（P<0.01）。结论 在三阴性乳腺癌细胞外泌体分泌过程中起核心作用的是RAB27a，抑制RAB27a能够抑制三阴性乳腺癌细胞的增殖、侵袭和粘附。

关键词: RAB27a；RAB27b；外泌体；三阴性乳腺癌；细胞生物学功能

Abstract: Objective To investigate the effect of inhibition of RAB27 protein family, which plays a pivotal role in exosome secretion, on biological behaviors of triple-negative breast cancer cells. Methods Quantitative real-time PCR and Western blotting were used to examine the expressions of RAB27 family and exosome secretion in 3 triple-negative breast cancer cell lines (MDA-MB-231, MDA-MB-468, and Hs578T) and a normal breast epithelial cell line (MCF10A). The effect of small interfering RNA (siRNA)-mediated silencing of RAB27a and RAB27b on exosome secretion in the 3 breast cancer cell lines was detected using Western blotting, and the changes in cell proliferation, invasion and adhesion were evaluated. Results Compared with normal breast epithelial cells, the 3 triple-negative breast cancer cell lines exhibited more active exosome secretion (P<0.001) and showed significantly higher expressions of RAB27a and RAB27b at both the mRNA and protein levels (P<0.01). Silencing of RAB27a in the breast cancer cells significantly down-regulated exosome secretion (P<0.001), while silencing of RAB27b did not significantly affect exosome secretion. The 3 breast cancer cell lines with RAB27a silencing-induced down-regulation of exosome secretion showed obvious inhibition of proliferation, invasion and adhesion (P<0.01) as compared with the cell lines with RAB27b silencing. Conclusion RAB27a plays central role in the exosome secretion in triple-negative breast cancer cells, and inhibiting RAB27a can inhibit the proliferation, invasion and adhesion of the cells.

Key words: RAB27a; RAB27b; exosomes; triple-negative breast cancer; cell biological behaviors