Wang Chun-Yan, Nie Qing-Wen, Zhou Xuan, Huang Da-Xiong, Xiao Wei-Qiang, Zhu Yong-Tong
Department of Neurology, Integrated Hospital of Traditional Chinese Medicine, Southern Medical University, Guangzhou 510315, China.E-mail:
Nan Fang Yi Ke Da Xue Xue Bao. 2017 Jul 20;37(7):869-874. doi: 10.3969/j.issn.1673-4254.2017.07.03.
To observe the effect of S100A4 gene silencing mediated by small interfering RNA (siRNA) on the proliferation of bladder cancer stem cells (CSCs) and their capacity of xenograft tumor formation.
MB49 bladder cancer stem cells (MCSCs) were isolated and identified. The differentially expressed protein S100A4 was identified in MCSCs using isobaric tags for relative and absolute quantitation technology (iTRAQ). A siRNA targeting S100A4 was constructed and transfected into MCSCs, and its inhibitory effects on S100A4 expression in MCSCs were assessed with Western blotting and qPCR. The effects of siRNA-mediated S100A4 silencing on the proliferation and xenograft tumor formation ability of MCSCs were observed.
Among the 65 differentially expressed proteins identified by iTRAQ combined with LC/MS/MS, S100A4 protein showed the most distinct differential expression in MCSCs. Transfection of MCSCs with S100A siRNA significantly inhibited the expressions of S100A4 at both mRNA and protein levels, caused obvious suppression of the cell proliferation, and attenuated the xenograft tumor formation ability of the cells in nude mice.
S100A4 in MCSCs is associated with the recurrence and metastasis of bladder cancer. S100A4 may serve as a potential therapeutic target for eliminating bladder CSCs.
观察小干扰RNA（siRNA）介导的S100A4基因沉默对膀胱癌干细胞（CSCs）增殖及其异种移植瘤形成能力的影响。
分离并鉴定MB49膀胱癌干细胞（MCSCs）。采用相对和绝对定量的等压标签技术（iTRAQ）在MCSCs中鉴定差异表达蛋白S100A4。构建靶向S100A4的siRNA并转染至MCSCs中，通过蛋白质印迹法和qPCR评估其对MCSCs中S100A4表达的抑制作用。观察siRNA介导的S100A4沉默对MCSCs增殖和异种移植瘤形成能力的影响。
通过iTRAQ联合液相色谱/串联质谱法鉴定出的65种差异表达蛋白中，S100A4蛋白在MCSCs中的差异表达最为明显。用S100A siRNA转染MCSCs可显著抑制S100A4在mRNA和蛋白水平的表达，明显抑制细胞增殖，并减弱细胞在裸鼠中的异种移植瘤形成能力。
MCSCs中的S100A4与膀胱癌的复发和转移相关。S100A4可能作为消除膀胱CSCs的潜在治疗靶点。
