本文選題：視網(wǎng)膜母細(xì)胞瘤 + ROCK蛋白��； 參考：《上海交通大學(xué)》2013年博士論文

【摘要】：研究目的和內(nèi)容:視網(wǎng)膜母細(xì)胞瘤（Retinoblastoma，RB）是小兒最常見(jiàn)的原發(fā)性眼內(nèi)惡性腫瘤，嚴(yán)重危害患兒的視力和生命，而RB的顱內(nèi)或全身轉(zhuǎn)移是RB患兒死亡的主要原因。有研究表明Rho/ROCK蛋白活性的改變與許多腫瘤細(xì)胞的增殖與侵襲密切相關(guān)，作為Rho的主要下游效應(yīng)蛋白，ROCK-1和ROCK-2對(duì)活體或離體腫瘤細(xì)胞的運(yùn)動(dòng)和侵襲能力有重要調(diào)節(jié)作用。但是，關(guān)于ROCK蛋白在RB細(xì)胞增殖與侵襲中的作用目前尚不清楚。 方法:采用RT-PCR和western blot方法檢測(cè)RB細(xì)胞株（Y79）中ROCK-1和ROCK-2的mRNA和蛋白的表達(dá)情況；應(yīng)用Lipofectamine2000將ROCK小分子干擾RNA (Small interfering RNA，siRNA)轉(zhuǎn)染到Y(jié)79細(xì)胞中，，觀(guān)察RNA干擾（RNA interference，RNAi）后細(xì)胞中的ROCK-1和ROCK-2的mRNA和蛋白的表達(dá)抑制情況；運(yùn)用ROCK抑制劑（Y-27632）和siRNA抑制ROCK蛋白的活性后，通過(guò)CCK-8細(xì)胞活性試驗(yàn)、matrigel細(xì)胞粘附試驗(yàn)和Transwell細(xì)胞侵襲試驗(yàn)，觀(guān)察ROCK蛋白對(duì)RB細(xì)胞增殖、轉(zhuǎn)移和侵襲能力的影響。 結(jié)果:實(shí)驗(yàn)結(jié)果顯示：在Y79細(xì)胞中有ROCK-1和ROCK-2mRNA和蛋白的表達(dá)，而且ROCK-1的表達(dá)要高于ROCK-2；siRNA介導(dǎo)的RNAi可以有效地抑制Y79細(xì)胞中ROCK-1和ROCK-2mRNA的表達(dá)，進(jìn)而抑制ROCK-1和ROCK-2蛋白的表達(dá)；ROCK抑制劑（Y-27632）對(duì)Y79細(xì)胞的增殖能力沒(méi)有影響，但可以顯著地增加Y79細(xì)胞的粘附能力并降低其侵襲能力；ROCK-1siRNA也可以顯著地增加Y79細(xì)胞的粘附能力并降低其侵襲能力，但ROCK-2siRNA對(duì)Y79細(xì)胞的粘附能力和侵襲能力沒(méi)有影響。 結(jié)論: ROCK-1蛋白在RB細(xì)胞轉(zhuǎn)移與侵襲中起重要作用，很有可能成為治療RB的新靶點(diǎn)。
[Abstract]:Objective and content: Retinoblastoma RBB is the most common primary intraocular malignant tumor in children, which seriously endangers the visual acuity and life of children. The intracranial or systemic metastasis of RB is the main cause of death in children with RB. Some studies have shown that the change of Rho/ROCK protein activity is closely related to the proliferation and invasion of many tumor cells. As the main downstream effector of Rho, Rock-1 and ROCK-2 play an important role in regulating the motion and invasiveness of tumor cells in vivo or in vitro. However, the role of ROCK protein in RB cell proliferation and invasion is unclear. Methods: RT-PCR and western blot were used to detect the expression of mRNA and protein of ROCK-1 and ROCK-2 in RB cell line Y79, and Lipofectamine2000 was used to transfect ROCK small interfering RNAs into Y79 cells. To observe the inhibition of mRNA and protein expression of ROCK-1 and ROCK-2 in the cells after RNA interference RNAi, and to observe the inhibition of ROCK protein activity by ROCK inhibitor, ROCK inhibitor, Y-27632) and siRNA. The cell adhesion test and Transwell cell invasion test were carried out by CCK-8 cell activity test. To observe the effect of ROCK protein on the proliferation, metastasis and invasion of RB cells. Results: the results showed that ROCK-1, ROCK-2mRNA and protein were expressed in Y79 cells, and the expression of ROCK-1 was higher than that of RNAi mediated by ROCK-2siRNA, which could effectively inhibit the expression of ROCK-1 and ROCK-2mRNA in Y79 cells. Furthermore, inhibition of the expression of ROCK-1 and ROCK-2 protein and the inhibition of the proliferation of Y79 cells by the inhibitor of rock Y-27632 did not affect the proliferation of Y79 cells. But the adhesion ability and invasion ability of Y79 cells were significantly increased and decreased by ROCK-2siRNA, but the adhesion and invasion ability of Y79 cells were not affected by ROCK-2siRNA. Conclusion: ROCK-1 protein plays an important role in the metastasis and invasion of RB cells and may be a new target for the treatment of RB.
【學(xué)位授予單位】：上海交通大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2013
【分類(lèi)號(hào)】：R739.7

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 ;Rho/ROCK pathway and neural regeneration: a potential therapeutic target for central nervous system and optic nerve damage[J];International Journal of Ophthalmology(English Edition);2011年06期

2 李永平,馮官光,易玉珍;國(guó)內(nèi)視網(wǎng)膜母細(xì)胞瘤的研究現(xiàn)狀及展望[J];中華眼科雜志;2004年04期

本文編號(hào)：1908233