本文選題：玫瑰樹(shù)堿 + 膀胱癌�。� 參考：《浙江大學(xué)》2017年博士論文

【摘要】：背景膀胱癌是我們國(guó)家泌尿系腫瘤發(fā)病率最高的惡性腫瘤。近年來(lái),膀胱癌的發(fā)病率還在呈現(xiàn)一種逐年上升的趨勢(shì)。膀胱癌的主要治療手段是手術(shù)和化療等。玫瑰樹(shù)堿是一種天然生物堿,玫瑰樹(shù)堿被證明在多種腫瘤中具有抑癌作用,但其對(duì)膀胱癌的作用效果及機(jī)制尚不明確。目的本研究通過(guò)體外實(shí)驗(yàn)探索玫瑰樹(shù)堿對(duì)膀胱癌T24細(xì)胞系增殖、周期、運(yùn)動(dòng)能力的影響和相應(yīng)的作用機(jī)制。方法玫瑰樹(shù)堿以不同濃度(l-16uM)和不同時(shí)間(24-72h)作用于T24細(xì)胞,采用MTT法和克隆形成實(shí)驗(yàn)檢測(cè)T24細(xì)胞在增殖和活性方面的改變,采用流式細(xì)胞術(shù)檢測(cè)細(xì)胞在周期方面的改變,采用RT-PCR實(shí)驗(yàn)檢測(cè)MMP,p53mRNA的表達(dá),采用siRNA技術(shù)干擾ATM的表達(dá),采用Transwell實(shí)驗(yàn)檢測(cè)細(xì)胞在運(yùn)動(dòng)能力方面的改變,采用Western Blot實(shí)驗(yàn)檢測(cè)相應(yīng)蛋白的改變。結(jié)果玫瑰樹(shù)堿以一種時(shí)間-劑量依賴(lài)的方式抑制膀胱癌T24細(xì)胞的增殖,表現(xiàn)為隨著濃度的升高、作用時(shí)間的延長(zhǎng),對(duì)細(xì)胞的抑制作用越強(qiáng)。玫瑰樹(shù)堿以濃度依賴(lài)的方式誘導(dǎo)T24細(xì)胞G2/M期周期阻滯,并分別上調(diào)和下調(diào)G2/M周期特異性蛋白cyclinB1和cdk1的表達(dá),同時(shí)能上調(diào)p53mRNA和蛋白的表達(dá)。玫瑰樹(shù)堿以濃度依賴(lài)的方式抑制膀胱癌T24細(xì)胞的遷移能力,并能下調(diào)MMP-9mRNA和蛋白的表達(dá)水平,同時(shí)上調(diào)E-cadherin的表達(dá)水平。玫瑰樹(shù)堿能上調(diào)ATM的磷酸化水平,上調(diào) Cdc25C(Ser-216)和 Chk1(Ser-345)的磷酸化水平。當(dāng) ATM 被 siRNA 干擾后,Cdc25C(Ser-216)和Chk1(Ser-345)的改變均出現(xiàn)了逆轉(zhuǎn)。結(jié)論玫瑰樹(shù)堿以濃度-時(shí)間依賴(lài)的方式抑制T24細(xì)胞的增殖能力,并誘導(dǎo)其G2/M期周期阻滯,抑制T24細(xì)胞的運(yùn)動(dòng)能力。玫瑰樹(shù)堿通過(guò)ATM-Chk1-Cdc25C-Cdk1和p53通路抑制T24細(xì)胞的增殖和誘導(dǎo)周期阻滯,通過(guò)EMT和MMPs抑制T24細(xì)胞的運(yùn)動(dòng)能力。
[Abstract]:Background bladder cancer is a malignant tumor with the highest incidence of urinary tract tumors in our country. In recent years, the incidence of bladder cancer is still showing a rising trend year by year. The main treatment of bladder cancer is surgery and chemotherapy. Rose tree alkaloid is a kind of natural alkaloid. Rose tree alkaloid has been proved to have anti-cancer effect in many kinds of tumors, but its effect on bladder cancer and its mechanism are not clear. Objective to investigate the effects of Roseine on the proliferation, cycle and motor ability of bladder cancer cell line T24 in vitro. Methods T24 cells were treated with roseine at different concentrations (l-16uM) and at different time (24-72 h). MTT assay and clone formation assay were used to detect the proliferation and activity of T24 cells. Flow cytometry was used to detect the changes of cell cycle. RT-PCR was used to detect the expression of MMPP p53 mRNA, siRNA technology was used to interfere with ATM expression, Transwell experiment was used to detect the change of cell motor ability, and Western Blot was used to detect the change of corresponding protein. Results Roseine inhibited the proliferation of bladder cancer T24 cells in a time-dose dependent manner. With the increase of concentration and the prolongation of action time, the inhibitory effect on T24 cells was stronger. Rosine induced G _ 2 / M phase arrest in T24 cells in a concentration-dependent manner, and up-regulated and down-regulated the expression of cyclin B1 and cdk1, and up-regulated the expression of p53 mRNA and protein, respectively. Rosine inhibited the migration of T24 cells in a concentration-dependent manner, down-regulated the expression of MMP-9 mRNA and protein, and up-regulated the expression of E-cadherin. Rose tree alkaloids can upregulate the phosphorylation level of ATM, CDc25C (Ser-216) and Chk1 (Ser-345). The changes of Cdc25C (Ser-216) and Chk1 (Ser-345) were reversed when ATM was interfered with siRNA. Conclusion Rosine inhibits the proliferation of T24 cells in a concentration-time dependent manner and induces G _ 2 / M cycle arrest and inhibits the motility of T _ 24 cells. Rosine inhibited the proliferation and cycle arrest of T24 cells through ATM-Chk1-Cdc25C-Cdk1 and p53 pathway, and inhibited the motor ability of T24 cells by EMT and MMPs.
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R285

【相似文獻(xiàn)】

相關(guān)期刊論文 前2條

1 徐兵河;劉云英;孫燕;;甲基羥基玫瑰樹(shù)堿治療晚期乳腺癌近期療效觀察[J];中國(guó)新藥雜志;1992年02期

2 ;[J];;年期

相關(guān)博士學(xué)位論文 前1條

1 陶水祥;玫瑰樹(shù)堿對(duì)人膀胱癌的作用及其機(jī)制的研究[D];浙江大學(xué);2017年

，

本文編號(hào)：2059254