S100A4:潜在抑制膀胱癌干细胞的靶分子

发布时间：2018-06-17 10:15

本文选题：膀胱癌干细胞 + SA　；参考：《南方医科大学学报》2017年07期

【摘要】：目的应用siRNA技术沉默S100A4基因表达对膀胱癌干细胞增殖及成瘤能力的影响。方法筛选鉴定出MB49膀胱癌干细胞后,应用核素标记相对和绝对定量(i TRAQ)技术,发现MB49膀胱癌细胞与膀胱癌干细胞表达差异的蛋白质S100A4蛋白。设计并合成S100A4基因特异性的siRNA序列,转染膀胱癌干细胞,应用Western blot和q PCR检测在siRNA对S100A4的影响,体内、外实验观察siRNA抑制S100A4后对膀胱癌干细胞增殖及成瘤能力的影响。结果通过i TRAQ核素标记结合液相色谱和串联质谱分析,共鉴定出差异蛋白共65个,S100A4在基因表达水平差异最显著(差异为5.70,P0.05)。与空白对照组、阴性对照组相比,S100A4 siRNA转染组的S100A4基因和蛋白表达降低(P0.05)。CCK8实验和裸鼠成瘤实验发现,与空白对照组与阴性对照组相比较,siRNA干扰S100A4表达后,膀胱癌干细胞生长明显受到抑制,成瘤能力受到抑制(P0.05)。结论膀胱癌干细胞中S100A4表达与膀胱癌的复发、转移有关,S100A4蛋白可能成为消灭膀胱癌干细胞,治疗膀胱癌的分子靶标。
[Abstract]:Objective to silence the expression of S100A4 gene by siRNA technique on the proliferation and tumorigenesis of bladder cancer stem cells. Methods after screening and identification of MB49 bladder cancer stem cells, the differential protein S100A4 protein was found between MB49 bladder cancer cells and bladder cancer stem cells. S100A4 gene specific siRNA sequence was designed and synthesized and transfected into bladder cancer stem cells. The effect of siRNA on S100A4 was detected by Western blot and Q PCR. In vitro and in vivo, the effects of siRNA on the proliferation and tumorigenesis of bladder cancer stem cells were observed. Results by means of iTRAQ radionuclide labeling, liquid chromatography and tandem mass spectrometry, 65 differentially expressed proteins (S100A4) were identified as the most significant difference in gene expression level (5.70% P0.05). Compared with the blank control group and the negative control group, the expression of S100A4 gene and protein in the S100A4 siRNA transfection group was lower than that in the control group, and the expression of S100A4 gene and protein in the control group was lower than that in the control group. CCK8 assay and tumorigenic assay in nude mice showed that siRNA interfered with the expression of S100A4 in comparison with the blank control group and the negative control group. The growth and tumorigenic ability of bladder cancer stem cells were inhibited significantly (P 0.05). Conclusion the expression of S100A4 in bladder cancer stem cells may be a molecular target for the elimination of bladder cancer stem cells and the treatment of bladder cancer.
【作者单位】：南方医科大学中西医结合医院脑病科;南方医科大学南方医院//第一临床医学院妇产科生殖中心;
【基金】：广东省自然科学基金(2016A030310403) 广东省医学科学技术研究基金(A2016092)
【分类号】：R737.14

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