本文選題：膀胱癌干細胞 + SA�。� 參考：《南方醫(yī)科大學(xué)學(xué)報》2017年07期

【摘要】：目的應(yīng)用siRNA技術(shù)沉默S100A4基因表達對膀胱癌干細胞增殖及成瘤能力的影響。方法篩選鑒定出MB49膀胱癌干細胞后,應(yīng)用核素標(biāo)記相對和絕對定量(i TRAQ)技術(shù),發(fā)現(xiàn)MB49膀胱癌細胞與膀胱癌干細胞表達差異的蛋白質(zhì)S100A4蛋白。設(shè)計并合成S100A4基因特異性的siRNA序列,轉(zhuǎn)染膀胱癌干細胞,應(yīng)用Western blot和q PCR檢測在siRNA對S100A4的影響,體內(nèi)、外實驗觀察siRNA抑制S100A4后對膀胱癌干細胞增殖及成瘤能力的影響。結(jié)果通過i TRAQ核素標(biāo)記結(jié)合液相色譜和串聯(lián)質(zhì)譜分析,共鑒定出差異蛋白共65個,S100A4在基因表達水平差異最顯著(差異為5.70,P0.05)。與空白對照組、陰性對照組相比,S100A4 siRNA轉(zhuǎn)染組的S100A4基因和蛋白表達降低(P0.05)。CCK8實驗和裸鼠成瘤實驗發(fā)現(xiàn),與空白對照組與陰性對照組相比較,siRNA干擾S100A4表達后,膀胱癌干細胞生長明顯受到抑制,成瘤能力受到抑制(P0.05)。結(jié)論膀胱癌干細胞中S100A4表達與膀胱癌的復(fù)發(fā)、轉(zhuǎn)移有關(guān),S100A4蛋白可能成為消滅膀胱癌干細胞,治療膀胱癌的分子靶標(biāo)。
[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.
【作者單位】： 南方醫(yī)科大學(xué)中西醫(yī)結(jié)合醫(yī)院腦病科;南方醫(yī)科大學(xué)南方醫(yī)院//第一臨床醫(yī)學(xué)院婦產(chǎn)科生殖中心;
【基金】：廣東省自然科學(xué)基金(2016A030310403) 廣東省醫(yī)學(xué)科學(xué)技術(shù)研究基金(A2016092)
【分類號】：R737.14

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