發(fā)布時間：2018-05-05 03:22

  本文選題：膀胱癌 + 吉西他濱　； 參考：《山東大學(xué)》2017年碩士論文

【摘要】：研究背景膀胱癌是西方國家和亞洲國家尿路上皮癌中最常見類型的癌癥,而且膀胱癌的流行率和死亡率逐年增加。它是美國第四大常見的癌癥,也是男性第九大常見的惡性腫瘤,在中國泌尿系統(tǒng)腫瘤的發(fā)病率中居首位。雖然隨著膀胱癌早期檢測手段的普及,患者5年生存率達到80%,但其5年復(fù)發(fā)率超過50%,并且會有10-20%的患者進展為浸潤性膀胱癌。吉西他濱做為膀胱癌的一線化療藥物已廣泛應(yīng)用于臨床,但是化療耐藥性的產(chǎn)生仍是不可避免的問題。因此,克服吉西他濱化療的耐藥性和提高化療的有效性,尋找克服化療耐藥性的新靶點和提高藥物的化療敏感性,已經(jīng)成為近期研究的熱點。提高吉西他濱有效的化療效率,并有效降低化療的毒性,能夠延長膀胱癌患者的生存時間,并改善預(yù)后,獲得更好的生活質(zhì)量。目的本研究首先探討了 CYLD和Livin與膀胱癌的進展之間的關(guān)系,然后通過穩(wěn)定轉(zhuǎn)染CYLD和Livin,研究CYLD和Livin在吉西他濱誘導(dǎo)下的對腫瘤細(xì)胞毒性、自噬及凋亡中的作用,為尋求改善癌癥細(xì)胞的化療敏感性,和促進減滅癌癥細(xì)胞的新的靶向分子目標(biāo),以及為當(dāng)前的癌癥化療方案尋找新的科研基礎(chǔ)。方法(1)培養(yǎng)膀胱癌細(xì)胞253J、T24,分別穩(wěn)定轉(zhuǎn)染CYLD、Livin,進行Western Blot、RT-PCR檢測各細(xì)胞株中CYLD及Livin的表達水平。(2)采用CCK-8檢測CYLD和Livin以及共轉(zhuǎn)染對膀胱癌細(xì)胞吉西他濱化療敏感性的影響,并計算其半數(shù)抑制濃度(50%inhibiting concentration,IC50);采用Transwell細(xì)胞侵襲及遷移實驗檢測CYLD和Livin抑制浸潤性膀胱癌細(xì)胞株253J、T24的侵襲及遷移能力。(3)通過Western Blot方法檢測聯(lián)合轉(zhuǎn)染后的T24及253J中NF-kB信號通路表達水平及自噬、凋亡相關(guān)蛋白的表達情況,探討聯(lián)合轉(zhuǎn)染中CYLD協(xié)同Livin誘導(dǎo)的對腫瘤細(xì)胞毒性及凋亡的分子通路機制,并通過string-db分析測CYLD和Livin的潛在關(guān)系。(4)建立小鼠皮下腫瘤模型,采用腹腔注射的方法將吉西他濱注射到小鼠體內(nèi),并將小鼠分組:對照組(DMSO)、CYLD組、Livin組、CYLD+Livin組。觀察并記錄各組對膀胱癌皮下腫瘤生長狀態(tài)的影響,分析CYLD和Livin對腫瘤模型增殖的抑制表現(xiàn)。結(jié)果(1)CCK-8細(xì)胞活性實驗檢測CYLD和Livin可分別提高膀胱癌細(xì)胞的化療敏感性,而CYLD聯(lián)合Livin可以大幅提高吉西他濱誘導(dǎo)的對腫瘤細(xì)胞毒性作用,進而促進了膀胱癌細(xì)胞對化療藥物吉西他濱的敏感性。(2)Transwell細(xì)胞實驗檢測CYLD和Livin可以使膀胱癌細(xì)胞株253J、T24的侵襲及遷移能力受到抑制,而CYLD聯(lián)合Livin的共轉(zhuǎn)染可以使膀胱癌細(xì)胞的侵襲、遷移能力受到明顯抑制。(3)Western Blot檢測膀胱癌細(xì)胞株253J、T24,發(fā)現(xiàn)膀胱癌細(xì)胞株吉西他濱化療敏感性增強可能與抑制細(xì)胞自噬蛋白LC3、P62,促進細(xì)胞凋亡蛋白Caspase3、Smac相關(guān),并發(fā)現(xiàn)CYLD可通過NF-kB信號通路調(diào)節(jié)Livin,進而提高癌細(xì)胞吉西他濱化療敏感性。(4)體內(nèi)試驗結(jié)果表明聯(lián)合轉(zhuǎn)染組(CYLD+Livin)能明顯增強吉西他濱對腫瘤的抑制作用。結(jié)論(1)過表達CYLD可以抑制膀胱癌細(xì)胞株的侵襲、遷移能力,提高其對吉西他濱的化療敏感性。(2)敲減Livin可以抑制膀胱癌細(xì)胞株的侵襲、遷移能力,提高其對吉西他濱的化療敏感性。(3)CYLD可通過NF-kB信號通路調(diào)節(jié)Livin(4)CYLD聯(lián)合Livin可明顯增加膀胱癌細(xì)胞在化療上對吉西他濱的敏感性,這種聯(lián)合治療為改進傳統(tǒng)的化療方案提高新的實驗基礎(chǔ)。
[Abstract]:Background bladder cancer is the most common type of cancer in the western and Asian countries, and the prevalence and mortality of bladder cancer are increasing year by year. It is the fourth most common cancer in the United States. It is also the ninth common malignant tumor in men. It ranks first in the incidence of urological tumors in China. Although with bladder cancer, it is the most common cancer in China. The 5 year survival rate of patients with early detection is 80%, but the recurrence rate of 5 years is more than 50%, and the patients with 10-20% will develop into invasive bladder cancer. Gemcitabine as a first-line chemotherapy drug for bladder cancer is widely used in clinical, but the production of chemotherapeutic drug resistance is still an inevitable problem. Therefore, to overcome gemcitabine To improve the efficiency of chemotherapy and reduce the toxicity of chemotherapy, it can prolong the survival time of the patients with bladder cancer and improve the prognosis, and get better prognosis. The purpose of this study was to investigate the relationship between CYLD and Livin and the progression of bladder cancer. Then through stable transfection of CYLD and Livin, the effects of CYLD and Livin on cytotoxicity, autophagy and apoptosis induced by gemcitabine, in order to improve chemosensitivity of cancer cells, and promote the reduction of cancer cells, were studied. New target molecular targets and new research foundation for current cancer chemotherapy scheme. Methods (1) 253J, T24, CYLD, Livin, Western Blot, RT-PCR were used to detect the expression level of CYLD and Livin in each cell line respectively. (2) CCK-8 CYLD and Livin, and co transfection of bladder cancer cells were used. The effect of chemosensitivity of Western itine, and the 50%inhibiting concentration, IC50, and Transwell cell invasion and migration test were used to detect the invasion and migration of CYLD and Livin in infiltrating bladder cancer cell line 253J, T24. (3) the Western Blot method was used to detect the T24 and 253J. The expression of autophagy and autophagy, the expression of autophagy and apoptosis related protein, and to explore the molecular pathway mechanism of CYLD synergistic Livin induced cytotoxicity and apoptosis in combined transfection, and the potential relationship between CYLD and Livin by string-db analysis. (4) to establish a mouse model of subcutaneous tumor and to injecting gemcitabine by intraperitoneal injection In mice, the mice were divided into groups: control group (DMSO), group CYLD, group Livin, group CYLD+Livin. The effects of each group on the growth of subcutaneous tumor of bladder cancer were observed and recorded, and the inhibition of CYLD and Livin on the proliferation of tumor model was analyzed. Results (1) CCK-8 cell activity testing CYLD and Livin can improve the sensitivity of bladder cancer cells to chemotherapy. CYLD combined with Livin can significantly increase the toxicity of gemcitabine on tumor cells and further promote the sensitivity of cystocarcinoma cells to the chemotherapeutic gemcitabine. (2) the detection of CYLD and Livin in Transwell cells can make the bladder cancer cell line 253J, the invasion and migration of T24, and CYLD combined with Livin. The transfection could inhibit the invasion and migration of bladder cancer cells. (3) Western Blot was used to detect the bladder cancer cell line 253J, T24. It was found that the chemosensitivity of the bladder cancer cell line gemcitabine may be associated with the inhibition of the autophagic protein LC3, P62, the promotion of apoptosis protein Caspase3, Smac, and the discovery of CYLD through NF-kB signals. Livin was used to improve the chemosensitivity of gemcitabine. (4) in vivo test results showed that the combined transfection group (CYLD+Livin) could significantly enhance the inhibitory effect of gemcitabine on tumor. Conclusion (1) overexpression of CYLD can inhibit the invasion and migration of bladder cancer cell lines and improve the chemosensitivity to gemcitabine. (2) knock down Li Vin can inhibit the invasion and migration of bladder cancer cell line and improve its chemosensitivity to gemcitabine. (3) CYLD can modulate Livin (4) CYLD combined with Livin through the NF-kB signaling pathway to significantly increase the sensitivity of bladder cancer cells to gemcitabine in chemotherapy. This combination therapy improves the traditional chemotherapy regimen to improve the new experiment Basics.

【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R737.14

【參考文獻】

相關(guān)期刊論文 前1條

1 ;The effect of C-terminal fragment of JNK2 on the stability of p53 and cell proliferation[J];Cell Research;2004年05期

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本文編號：1845968