本文選題：膀胱癌 + miR-143�。� 參考：《南京醫(yī)科大學(xué)》2015年博士論文

【摘要】：背景:miR-143是microRNA的一種,已被證實(shí)在多種腫瘤組織中低表達(dá),并可以起到抑癌基因的作用,體外細(xì)胞實(shí)驗(yàn)中證實(shí)IGF-IR3'-UTR區(qū)存在miR-143調(diào)控位點(diǎn),高表達(dá)的miR-143能作用于IGF-IR來(lái)抑制腫瘤細(xì)胞增殖、侵襲、轉(zhuǎn)移。microRNA在基因調(diào)控中所起的作用受到越來(lái)越多的重視,有關(guān)在膀胱癌細(xì)胞中miR-143調(diào)控IGF-IR影響吉西他濱化療敏感性的研究尚未見(jiàn)報(bào)道。我們的研究提示miR-143可能在膀胱癌細(xì)胞的IGF-IR基因調(diào)控機(jī)制中起到重要的作用,探索miR-143介導(dǎo)的IGF-IR基因調(diào)控作用對(duì)膀胱癌細(xì)胞生物學(xué)行為和化療藥物敏感性的影響十分重要。目的:明確miR-143在膀胱癌組織標(biāo)本和細(xì)胞中的表達(dá)水平及其與膀胱癌細(xì)胞生長(zhǎng)的相關(guān)性,探討miR-143影響膀胱癌惡性生物學(xué)行為和對(duì)化療藥物敏感性的分子機(jī)制。方法:1、RT-PCR方法檢測(cè)23例膀胱癌及其相對(duì)應(yīng)的癌旁正常膀胱組織標(biāo)本中miR-143的表達(dá)水平,并檢測(cè)膀胱癌細(xì)胞株T24和5637細(xì)胞中miR-143的表達(dá)水平,分析miR-143表達(dá)與膀胱癌分期的相關(guān)性。2、RT-PCR方法檢測(cè)23例膀胱癌及其相對(duì)應(yīng)的癌旁正常膀胱組織標(biāo)本{中IGF-IR的表達(dá)水平,分析miR-143與IGF-IR表達(dá)的相關(guān)性。構(gòu)建miR-143過(guò)表達(dá)慢病毒系統(tǒng),轉(zhuǎn)染膀胱癌細(xì)胞。RT-PCR方法檢測(cè)過(guò)表達(dá)miR-143的T24和5637細(xì)胞中IGF-IR的表達(dá)水平。3、CCK-8法檢測(cè)過(guò)表達(dá)miR-143和干擾IGF-IR對(duì)5637細(xì)胞增殖的影響。同時(shí)檢測(cè)過(guò)表達(dá)miR-143和干擾IGF-IR對(duì)吉西他濱敏感性影響。4、Western blotting 實(shí)驗(yàn)分析過(guò)表達(dá) miR-143 和干擾 IGF-IR 后 p-AKT、AKT、p-ERK和ERK的蛋白表達(dá)情況。結(jié)果:1、23對(duì)膀胱癌組織標(biāo)本中,miR-143在膀胱癌組織中表達(dá)低于癌旁正常膀胱組織,并且其表達(dá)水平與腫瘤的分期密切相關(guān),在膀胱癌細(xì)胞株T24和5637中miR-143表達(dá)降低。2、miR-143和IGF-IR在膀胱癌組織標(biāo)本中的表達(dá)呈負(fù)相關(guān),在T24和5637細(xì)胞中高表達(dá)的miR-143能明顯抑制IGF-IR表達(dá)。3、在5637細(xì)胞中過(guò)表達(dá)miR-143和干擾高表達(dá)IGF-IR均能抑制細(xì)胞的生長(zhǎng)。過(guò)表達(dá)miR-143和干擾高表達(dá)IGF-IR的5637細(xì)胞對(duì)吉西他濱的敏感性增加。4、上調(diào)miR-143能抑制5637細(xì)胞中P-AKT和P-ERK的表達(dá)。干擾5637細(xì)胞中IGF-IR表達(dá)可以調(diào)節(jié)AKT和ERK信號(hào)通路的激活來(lái)抑制P-AKT和P-ERK的表達(dá)。結(jié)論:1、miR-143在膀胱癌組織和細(xì)胞中廣泛低表達(dá),與膀胱癌的惡性生物學(xué)行為密切相關(guān)。2、miR-143抑制膀胱癌細(xì)胞的增殖能力,并且可以通過(guò)靶向IGF-IR影響AKT和ERK信號(hào)通路起到抑癌基因的作用。3、miR-143和吉西他濱對(duì)膀胱癌細(xì)胞生長(zhǎng)有協(xié)同抑制作用,miR-143可以增加膀胱癌細(xì)胞對(duì)化療藥物(吉西他濱)的敏感性。
[Abstract]:Background: miR-143 is a kind of microRNA, which has been proved to be low expressed in many kinds of tumor tissues and can act as a tumor suppressor gene. In vitro, it has been proved that there are miR-143 regulatory sites in the IGF-IR3'-UTR region, and that high expression miR-143 can act on IGF-IR to inhibit the proliferation of tumor cells. More and more attention has been paid to the role of invasion, metastasis, microRNAs in gene regulation. The effect of miR-143 on the chemosensitivity of gemcitabine in bladder cancer cells has not been reported. Our results suggest that miR-143 may play an important role in the regulation of IGF-IR gene in bladder cancer cells. It is important to explore the effects of miR-143 mediated IGF-IR gene regulation on the biological behavior and chemosensitivity of bladder cancer cells. Objective: to investigate the expression of miR-143 in bladder cancer tissues and its correlation with the growth of bladder cancer cells, and to explore the molecular mechanism of miR-143 affecting malignant biological behavior and chemosensitivity of bladder cancer. Methods the expression of miR-143 in 23 cases of bladder cancer and its adjacent normal bladder tissues was detected by using 1: 1 RT-PCR, and the expression of miR-143 in bladder cancer cell lines T24 and 5637 was also detected. To analyze the correlation between the expression of miR-143 and the stage of bladder cancer. 2RT-PCR was used to detect the expression of IGF-IR in 23 cases of bladder cancer and its adjacent normal bladder tissues, and to analyze the correlation between miR-143 and IGF-IR expression. MiR-143 overexpression lentivirus system was constructed. The expression level of IGF-IR in T24 and 5637 cells overexpressed with miR-143 was detected by RT-PCR. The effect of overexpression of miR-143 and interference of IGF-IR on the proliferation of 5637 cells was detected by the method of CCK-8. The effect of overexpression of miR-143 and interference of IGF-IR on the sensitivity of gemcitabine. 4Western blotting assay was used to analyze the expression of p-AKTT-AK p-ERK and ERK after IGF-IR interference and overexpression of miR-143. Results the expression of miR-143 in bladder cancer tissues was lower than that in adjacent normal bladder tissues, and the expression level was closely related to the stage of the tumor. The expression of miR-143 and IGF-IR in bladder cancer cell lines T24 and 5637 were negatively correlated. Overexpression of miR-143 in T24 and 5637 cells significantly inhibited the expression of IGF-IR. 3. Overexpression of miR-143 and interference of overexpression of IGF-IR in 5637 cells inhibited the growth of T24 and 5637 cells. Overexpression of miR-143 and overexpression of IGF-IR increased the sensitivity of 5637 cells to gemcitabine. Upregulation of miR-143 inhibited the expression of P-AKT and P-ERK in 5637 cells. Interfering with the expression of IGF-IR in 5637 cells can inhibit the expression of P-AKT and P-ERK by regulating the activation of AKT and ERK signaling pathway. Conclusion the low expression of 1: 1 miR-143 in bladder cancer tissues and cells is closely related to the malignant biological behavior of bladder cancer. It can inhibit the proliferation of bladder cancer cells. Moreover, targeting IGF-IR could inhibit the growth of bladder cancer cells by targeting AKT and ERK signaling pathway. 3miR-143 and gemcitabine had synergistic inhibitory effects on the growth of bladder cancer cells. MiR-143 could increase the sensitivity of bladder cancer cells to chemotherapeutic drugs (gemcitabine).
【學(xué)位授予單位】：南京醫(yī)科大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2015
【分類號(hào)】：R737.14

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本文編號(hào)：1905984