本文選題：TRIM3 + 肝癌　； 參考：《廣東藥學(xué)院》2015年碩士論文

【摘要】：研究背景及目的肝癌(hepatocellular carcinoma,HCC)是一種全球常見(jiàn)的惡性腫瘤,在所有實(shí)體腫瘤中發(fā)病率居第五位,腫瘤相關(guān)死亡率居于第四位。我國(guó)為原發(fā)性肝癌的高發(fā)區(qū),近幾年呈現(xiàn)發(fā)病率上升的趨勢(shì)。盡管目前肝癌的治療手段有了很大程度的進(jìn)步,但是肝癌預(yù)后并不理想,五年生存率只有25%-39%。導(dǎo)致肝癌預(yù)后差的主要原因是惡性程度高、血管侵犯及肝內(nèi)轉(zhuǎn)移。肝癌的發(fā)病過(guò)程是多種因素共同作用的結(jié)果,其中遺傳因素中最主要的是癌基因激活和抑癌基因失活。為了改善肝癌患者預(yù)后,有必要尋找新的早期診斷和有效治療肝癌的分子靶點(diǎn)。TRIM3蛋白又稱BERP蛋白,最初是作為作為α-輔肌動(dòng)蛋白4結(jié)合蛋白而被克隆出來(lái),但是它的具體功能目前并不是十分明確。TRIM家族蛋白在結(jié)構(gòu)上均有功能相似的3個(gè)結(jié)構(gòu)域(靠近N端的鋅指結(jié)構(gòu)域、位于中間的1-2個(gè)B-box結(jié)構(gòu)域、C端的卷曲螺旋結(jié)構(gòu)域),也因此而得名。近年來(lái)有多項(xiàng)研究證實(shí)TRIM家族蛋白成員與腫瘤的發(fā)生發(fā)展密切相關(guān),有研究證實(shí)在惡性膠質(zhì)瘤中TRIM3是一個(gè)新的抑癌基因,TRIM3發(fā)揮其抑制細(xì)胞生長(zhǎng)的作用是通過(guò)抑制p21蛋白表達(dá)而起作用。TRIM3表達(dá)下調(diào)能促進(jìn)細(xì)胞中p21的表達(dá)增加,并且能促進(jìn)細(xì)胞增殖和小鼠腫瘤的生長(zhǎng)和血管形成。過(guò)表達(dá)TRIM3可使細(xì)胞在細(xì)胞周期G0/1期累積,導(dǎo)致細(xì)胞生長(zhǎng)阻滯。因此認(rèn)為T(mén)RIM3可能是一個(gè)潛在的抑癌基因,然而,到目前為止,TRIM3在肝癌中的作用仍不清楚。本課題的目的是通過(guò)檢測(cè)TRIM3基因在肝癌中的表達(dá),分析其與腫瘤患者臨床參數(shù)及預(yù)后的關(guān)系,并初步研究TRIM3在肝癌的發(fā)生過(guò)程當(dāng)中的生物學(xué)功能。研究方法實(shí)時(shí)熒光定量RT-PCR、Western Blotting和免疫組織化學(xué)技術(shù)檢測(cè)肝癌細(xì)胞系及肝癌組織標(biāo)本中TRIM3的表達(dá)情況。通過(guò)轉(zhuǎn)染TRIM3基因片段,使TRIM3蛋白在Hep G2和Bel-7402中過(guò)表達(dá),研究TRIM3的表達(dá)對(duì)肝癌細(xì)胞生長(zhǎng)(增殖、克隆形成)、細(xì)胞周期、細(xì)胞凋亡、細(xì)胞轉(zhuǎn)移(遷移及侵襲)等生物學(xué)功能的影響。統(tǒng)計(jì)方法:配對(duì)t檢驗(yàn)主要用于肝癌組織與癌旁正常組織TRIM3轉(zhuǎn)錄和蛋白表達(dá)的差異比較;卡方檢驗(yàn)主要用于高表達(dá)組和低表達(dá)組臨床參數(shù)之間的差異比較;生存分析用于比較高表達(dá)組和低表達(dá)組生存時(shí)間的比較;Cox回歸分析主要用于研究與肝癌術(shù)后總體生存時(shí)間相關(guān)的影響因素。結(jié)果1.肝癌組織中TRIM3 m RNA和蛋白的表達(dá)水平顯著低于配對(duì)的癌旁組織(p0.05)。與肝正常細(xì)胞系LO2相比,TRIM3在肝癌細(xì)胞株中的表達(dá)下調(diào),尤其是在Hep G2和Bel7402這兩株肝癌細(xì)胞中。2.免疫組織化學(xué)法檢測(cè)了129例石蠟包埋組織中TRIM3蛋白的表達(dá)情況,發(fā)現(xiàn)TRIM3蛋白主要表達(dá)在細(xì)胞漿中,相對(duì)于癌組織的低表達(dá),癌旁的肝組織為高表達(dá);TRIM3表達(dá)與腫瘤大小(P=0.034)、病理分級(jí)(P0.001)、TNM分期(P=0.021)和血清AFP水平(P=0.025)顯著相關(guān);生存分析表明TRIM3低表達(dá)的患者預(yù)后不良,可作為肝癌患者獨(dú)立的預(yù)后指標(biāo)。3.構(gòu)建過(guò)表達(dá)TRIM3的慢病毒表達(dá)載體,感染Hep G2和Bel7402細(xì)胞,發(fā)現(xiàn)過(guò)表達(dá)TRIM3能抑制肝癌細(xì)胞的增殖、克隆形成、以及遷移侵襲能力,誘導(dǎo)細(xì)胞周期阻滯于G1期,促進(jìn)腫瘤細(xì)胞的凋亡。結(jié)論TRIM3在肝癌的發(fā)生發(fā)展中具有重要作用,可以作為肝癌分子靶向治療的靶點(diǎn)和肝癌預(yù)后的一個(gè)獨(dú)立指標(biāo)。
[Abstract]:Background and objective hepatocellular carcinoma (HCC) is one of the most common malignant tumors in the world. The incidence of cancer is fifth in all solid tumors and fourth in tumor related mortality. China is a high incidence area of primary liver cancer in China. The prognosis of liver cancer is not good. The prognosis of liver cancer is not ideal. The main cause of poor prognosis of the five year survival rate is high malignancy, vascular invasion and intrahepatic metastasis. The pathogenesis of liver cancer is the result of a variety of factors, of which the most important of the genetic factors is the activation of oncogene and the inactivation of tumor suppressor genes. The prognosis of good liver cancer patients, it is necessary to find new early diagnosis and effective treatment of liver cancer molecular target.TRIM3 protein called BERP protein, which was originally cloned as alpha cofactor 4 binding protein, but its specific function is not very clear about the 3 knots of the.TRIM family protein in structure. The domain (the domain of the zinc finger near the N, 1-2 B-box domains in the middle, the curl spiral domain in the C end) is also named. In recent years, a number of studies have confirmed that the members of the TRIM family protein are closely related to the development of the tumor. Studies have shown that TRIM3 is a new tumor suppressor gene in the malignant glioma, and TRIM3 plays its inhibition. The role of cell growth is to inhibit the expression of p21 protein by inhibiting the expression of.TRIM3, which can promote the increase of p21 expression in cells, and promote cell proliferation and tumor growth and angiogenesis in mice. Overexpression of TRIM3 can cause cells to accumulate in the G0/1 phase of cell cycle, leading to cell growth block. Therefore, TRIM3 may be a possible one. Potential tumor suppressor genes, however, the role of TRIM3 in liver cancer is still unclear so far. The purpose of this study is to analyze the relationship between the expression of TRIM3 gene in liver cancer, analyze the relationship with the clinical parameters and prognosis of the cancer patients, and preliminary study the biological function of TRIM3 in the process of liver cancer. Quantitative RT-PCR, Western Blotting and immunohistochemical technique were used to detect the expression of TRIM3 in liver cancer cell lines and hepatocellular carcinoma tissue specimens. By transfecting TRIM3 gene fragments, TRIM3 protein was overexpressed in Hep G2 and Bel-7402, and the expression of TRIM3 was studied for the growth of hepatoma cells (proliferation, clone formation), cell cycle, cell apoptosis and cell transformation. The effects of biological functions such as migration and invasion. Statistical methods: the paired t test was used to compare the difference in the TRIM3 transcription and protein expression between the liver cancer tissue and the paracancerous normal tissues; the chi square test was mainly used to compare the differences between the high expression group and the low expression group, and the survival analysis was used to compare the high expression group and the low expression group. Comparison of survival time; Cox regression analysis was mainly used to study the influence factors related to the overall survival time after liver cancer. Results 1. the expression level of TRIM3 m RNA and protein in liver cancer tissues was significantly lower than that of the paired paracancerous tissue (P0.05). The expression of TRIM3 in the hepatocellular carcinoma cell lines was down, especially in Hep G, compared with the normal liver cell line LO2. The expression of TRIM3 protein in 129 paraffin embedded tissues was detected by.2. immunohistochemistry in 2 and Bel7402 hepatoma cells. The expression of TRIM3 protein was mainly expressed in the cytoplasm. The expression of the liver tissue near the cancer tissue was highly expressed, the expression of TRIM3 and the size of the tumor (P=0.034), the pathological grade (P0.001), and the TNM staging (P=0). .021) was significantly related to serum AFP level (P=0.025); survival analysis showed that the patients with low expression of TRIM3 had poor prognosis, and could be used as an independent prognostic indicator of liver cancer patients.3. to construct a lentivirus expression vector expressing TRIM3, infected with Hep G2 and Bel7402 cells, and that overexpressed TRIM3 could inhibit the proliferation, clone formation, and migration invasion of hepatoma cells. The ability to induce cell cycle arrest in G1 phase and promote the apoptosis of tumor cells. Conclusion TRIM3 plays an important role in the development and development of liver cancer. It can be used as a target for the target therapy of liver cancer and an independent indicator of the prognosis of liver cancer.
【學(xué)位授予單位】：廣東藥學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類(lèi)號(hào)】：R735.7

【參考文獻(xiàn)】

相關(guān)期刊論文 前3條

1 倪曉光;趙平;;泛素-蛋白酶體途徑與惡性腫瘤關(guān)系的研究進(jìn)展[J];中華腫瘤防治雜志;2007年19期

2 田利源;陳紅星;鄧?yán)^先;;一個(gè)新的與泛素化有關(guān)的蛋白家族——TRIM家族[J];生物技術(shù)通訊;2007年02期

3 吳怡;劉新;;蛋白質(zhì)泛素化、去泛素化與腫瘤發(fā)生的關(guān)系[J];沈陽(yáng)醫(yī)學(xué)院學(xué)報(bào);2008年01期

相關(guān)會(huì)議論文 前1條

1 韓雪琳;閆東梅;;TRIM家族的研究進(jìn)展[A];第八屆全國(guó)免疫學(xué)學(xué)術(shù)大會(huì)論文集[C];2012年

，

本文編號(hào)：1939989