發(fā)布時(shí)間：2021-12-29 00:02

　　目的:探討TGF-β1抑制的microRNAs及其靶基因蛋白通過MAPK信號(hào)通路調(diào)節(jié)人類橫紋肌肉瘤（RMS）肌分化及其分子機(jī)制,為RMS分化治療提供新的治療靶點(diǎn)。方法:（1）采用Real-time PCR技術(shù)檢測TGF-β1抑制的microRNAs在不同RMS細(xì)胞株和組織樣本中的表達(dá),篩選出差異最明顯、最廣泛的microRNA作為研究對象。（2）利用MTT、3H胸腺嘧啶核苷摻入法檢測microRNA對RMS細(xì)胞株增殖的影響。（3）構(gòu)建裸鼠成瘤性模型,觀察microRNA對活體腫瘤生長的影響;采用免疫組織化學(xué)染色檢測瘤體ki-67蛋白表達(dá)情況。（4）運(yùn)用生物信息學(xué)技術(shù)及網(wǎng)絡(luò)資源預(yù)測miR-411-5p的靶基因（5）運(yùn)用Real-time PCR技術(shù)、雙熒光素酶報(bào)告系統(tǒng)及蛋白印跡法驗(yàn)證靶基因。（6）構(gòu)建Elk-1-Luc、c-Jun-Luc熒光報(bào)告質(zhì)粒,SPRY4-siRNA或siRNA-ctrl轉(zhuǎn)染PKC?表達(dá)質(zhì)粒處理的RMS細(xì)胞株,檢測Elk-1-Luc、c-Jun-Luc熒光信號(hào);PKC?表達(dá)質(zhì)粒和SPRY4-siRNA/siRNA-ctrl共轉(zhuǎn)染RMS細(xì)胞株,檢測P-p38蛋白的變... 

【文章來源】：蘇州大學(xué)江蘇省 211工程院校

【文章頁數(shù)】：83 頁

【學(xué)位級(jí)別】：碩士

【部分圖文】：

驗(yàn)證TGF-β1基因敲低效果

TGF-β1/miR-411-5p/SPRY4和 MAPK信號(hào)通路參與橫紋肌肉瘤分化調(diào)表達(dá)明顯增加 (圖 2 )。miR-411-5p 和 miR-411-3p 在 RMS、更普遍，因此選擇 miR-411-5p 在 RMS 組織中進(jìn)一步驗(yàn)

RT-PCR檢測miR-411-5p六對低/高TGF-β1表達(dá)的ARMS和八對ERMS組織標(biāo)本

【參考文獻(xiàn)】：
期刊論文
[1]Antisense Oligonucleotide Targeting TGF-β1 Abrogates Tumorigenicity of Rhabdomyosarcoma in vivo[J]. Shouli Wang Lingling Guo Liang Dong Shigang Li Haizhen Deng Department of Pathology,Medical School of Soochow University,Suzhou 215123,Jiangsu Province,China. Huihua Yao Department of Surgery,the Second Hospital of Soochow University,Suzhou 215123,Jiangsu Province,China. Maomin Sun Department of Anatomy,Medical School of Soochow University,Suzhou 215123,Jiangsu Province.China..  Chinese Journal of Clinical Oncology. 2008(04)
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