Wang L, Yan Z, Xia Y
Department of Radiation Oncology, Yunnan Provincial Cancer Hospital (Third Affiliated Hospital of Kunming Medical University), Kunming 650118, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2023 Apr 20;43(4):560-567. doi: 10.12122/j.issn.1673-4254.2023.04.08.
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.
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.
Compared with normal breast epithelial cells, the 3 triple-negative breast cancer cell lines exhibited more active exosome secretion ( < 0.001) and showed significantly higher expressions of RAB27a and RAB27b at both the mRNA and protein levels ( < 0.01). Silencing of RAB27a in the breast cancer cells significantly down-regulated exosome secretion ( < 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 ( < 0.01) as compared with the cell lines with RAB27b silencing.
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.
研究在细胞外泌体分泌中起关键作用的RAB27蛋白家族的抑制对三阴性乳腺癌细胞生物学行为的影响。
采用定量实时聚合酶链反应和蛋白质免疫印迹法检测3种三阴性乳腺癌细胞系(MDA-MB-231、MDA-MB-468和Hs578T)及1种正常乳腺上皮细胞系(MCF10A)中RAB27家族的表达及细胞外泌体分泌情况。利用蛋白质免疫印迹法检测小干扰RNA介导的RAB27a和RAB27b沉默对3种乳腺癌细胞系细胞外泌体分泌的影响,并评估细胞增殖、侵袭和黏附的变化。
与正常乳腺上皮细胞相比,3种三阴性乳腺癌细胞系表现出更活跃的细胞外泌体分泌(<0.001),且在mRNA和蛋白质水平上RAB27a和RAB27b的表达均显著更高(<0.01)。乳腺癌细胞中RAB27a的沉默显著下调了细胞外泌体分泌(<0.001),而RAB27b的沉默对细胞外泌体分泌无显著影响。与RAB27b沉默的细胞系相比,3种因RAB27a沉默导致细胞外泌体分泌下调的乳腺癌细胞系在增殖、侵袭和黏附方面均表现出明显抑制(<0.01)。
RAB27a在三阴性乳腺癌细胞的细胞外泌体分泌中起核心作用,抑制RAB27a可抑制细胞的增殖、侵袭和黏附。