本文關(guān)鍵詞： 結(jié)直腸癌 IQGAP3 增殖 遷移 KRAS　出處：《南方醫(yī)科大學(xué)》2017年碩士論文　論文類(lèi)型：學(xué)位論文

【摘要】：研究背景和目的:結(jié)直腸癌(Colorectal Cancer,CRC)是世界范圍內(nèi)最常見(jiàn)的惡性腫瘤之一,其發(fā)病率位居所有惡性腫瘤第三位,死亡率位居第四位,僅次于肺癌、肝癌和胃癌。在中國(guó),CRC的發(fā)病率和死亡率自2010年以來(lái)一直在持續(xù)增長(zhǎng),成為癌癥患者死亡的主要原因,并呈現(xiàn)年輕化趨勢(shì)。CRC不僅降低了患者的生活質(zhì)量,也給社會(huì)帶來(lái)了沉重的經(jīng)濟(jì)負(fù)擔(dān);然而,CRC發(fā)生和發(fā)展的分子機(jī)制目前尚不完全清楚。CRC的發(fā)生是一個(gè)多階段、多步驟的復(fù)雜過(guò)程,遵循著“正常-增生-腺瘤-癌”等發(fā)展變化過(guò)程;主要機(jī)制有包括染色體不穩(wěn)定(CNI)、微衛(wèi)星不穩(wěn)定(MSI)、CpG島甲基化表型(CIMP)、鋸齒狀等發(fā)生途徑,還有一部分CRC的發(fā)生發(fā)展涉及兩條及以上方式。腫瘤轉(zhuǎn)移是臨床上導(dǎo)致患者死亡的主要原因,其過(guò)程主要包括局部浸潤(rùn)、滲入血管、隨血循環(huán)流動(dòng)、穿出血管,最終在遠(yuǎn)處器官定植并形成新的轉(zhuǎn)移灶等過(guò)程,在這一系列過(guò)程中,涉及了許多復(fù)雜的因素。所以,深入研究CRC發(fā)生、發(fā)展和侵襲轉(zhuǎn)移的分子機(jī)制,尋找有價(jià)值的分子標(biāo)志物,對(duì)CRC的早期診斷、預(yù)后評(píng)估以及靶向治療等均有重要科學(xué)意義和臨床價(jià)值。IQGAP3基因位于染色體1q21.3,分子量大小約為180KD,含1631個(gè)氨基酸,主要在腸道、肝臟及腦、肺等器官中表達(dá)。研究發(fā)現(xiàn),IQGAP3能夠通過(guò)特定的結(jié)構(gòu)區(qū)結(jié)合相關(guān)蛋白質(zhì),影響細(xì)胞增殖、分化、細(xì)胞粘附、細(xì)胞骨架以及細(xì)胞運(yùn)動(dòng)等。目前對(duì)IQGAP3的研究主要集中在神經(jīng)突增生、細(xì)胞骨架形成等方面,研究還發(fā)現(xiàn)IQGAP3表達(dá)紊亂還參與肝癌、肺癌等惡性腫瘤的發(fā)生和發(fā)展等過(guò)程;然而,其表達(dá)失調(diào)在CRC發(fā)生和發(fā)展過(guò)程中的作用及分子機(jī)制目前尚不清楚。因此,本研究擬深入探討IQGAP3在CRC發(fā)生和演進(jìn)中的功能和分子機(jī)制。方法1.利用TCGA公共數(shù)據(jù)庫(kù)、免疫組化(Immunohistochemistry,IHC)法分析、檢測(cè)CRC組織及相應(yīng)癌旁正常粘膜組織中IQGAP3的表達(dá)情況;2.利用熒光實(shí)時(shí)定量PCR、蛋白免疫印跡法檢測(cè)IQGAP3在腸癌組織和細(xì)胞株中的表達(dá),并構(gòu)建IQGAP3穩(wěn)定過(guò)表達(dá)和干擾細(xì)胞株;3.采用MTT、平板克隆形成、軟瓊脂克隆形成實(shí)驗(yàn)和裸鼠皮下成瘤實(shí)驗(yàn),劃痕愈合實(shí)驗(yàn)、Transwell小室實(shí)驗(yàn),檢測(cè)IQGAP3對(duì)腫瘤細(xì)胞增殖和遷移能力的影響;4.通過(guò)GSEA基因富集分析預(yù)測(cè)IQGAP3參與的生物過(guò)程,利用流式細(xì)胞術(shù)及Western blot的方法檢測(cè)IQGAP3過(guò)表達(dá)或被干擾后相關(guān)信號(hào)通路靶基因的變化情況,探討IQGAP3參與CRC發(fā)生和遷移的分子機(jī)制。結(jié)果1.IQGAP3在結(jié)直腸癌組織中的表達(dá)水平高于相配對(duì)的癌旁正常組織;2.IQGAP3在結(jié)直腸癌細(xì)胞株中的表達(dá)高于正常腸粘膜細(xì)胞;3.IQGAP3過(guò)表達(dá)能夠顯著促進(jìn)CRC細(xì)胞的增殖及遷移能力,而IQGAP3被干擾后能夠顯著抑制CRC細(xì)胞的增殖及遷移能力;4.IQGAP3影響KRAS信號(hào)通路的活性及其下游靶基因的表達(dá)。結(jié)論1.IQGAP3在CRC組織中的表達(dá)水平顯著高于配對(duì)的正常腸黏膜組織;2.IQGAP3過(guò)表達(dá)能夠顯著促進(jìn)CRC細(xì)胞的增殖及遷移能力,而IQGAP3被干擾后能夠顯著抑制CRC細(xì)胞的增殖及遷移能力;3.IQGAP3通過(guò)調(diào)控KRAS/AKT信號(hào)通路、KRAS/Rac1、CDC42信號(hào)通路,促進(jìn)CRC細(xì)胞的增殖和遷移。
[Abstract]:Background and objective: colorectal cancer (Colorectal, Cancer, CRC) is one of the most common malignant tumors in the world, the incidence rate of third among all malignant tumors, the mortality rate ranks fourth only to lung cancer, liver cancer and gastric cancer. In China, the incidence and mortality of CRC since 2010 has continued to grow, become the main cause of death in cancer patients, and there is a trend of young.CRC not only reduces the patient's quality of life, also brought a heavy economic burden; however, the molecular mechanism of the occurrence and development of CRC is not entirely clear to the occurrence of.CRC is a multi-stage, multi-step complex process, follow the "development changes in normal hyperplasia adenoma carcinoma" process; including the main mechanism of chromosomal instability (CNI) and microsatellite instability (MSI), CpG island methylator phenotype (CIMP), zigzag way, and a The occurrence of CRC involved in the development of two and above. The clinical tumor metastasis is the leading cause of death in patients, which mainly includes local infiltration, infiltration of blood flow, the blood circulation, through blood vessels, eventually in distant organs colonization and the formation of new metastases in this process, a series of process. Involves many complicated factors. Therefore, in-depth study of the molecular mechanism of CRC occurrence, development and metastasis and find valuable molecular markers for early diagnosis, CRC, prognosis and targeted therapy has important scientific significance and clinical value of.IQGAP3 gene is located on chromosome 1q21.3, the molecular weight is about 180KD, including 1631 amino acids, mainly in the intestine, liver and brain, the expression of lung and other organs. The study found that IQGAP3 was able to structure specific binding proteins, adhesion affect cell proliferation, differentiation, cells, bone cells Frame and cell movement. The current research on IQGAP3 mainly focus on the proliferation of neurites, cytoskeleton formation and other aspects, the study also found that IQGAP3 expression disorder is also involved in the occurrence and development of liver cancer, lung cancer and other malignant tumors; however, its expression is dysregulated in CRC incidence and effects and the molecular mechanisms in the development process is still not clear. Therefore, this study intends to explore the function of IQGAP3 and CRC in the molecular mechanism of carcinogenesis and progression. Methods 1. using the TCGA public database, immunohistochemistry (Immunohistochemistry, IHC) analysis to detect expression of IQGAP3 CRC tissues and adjacent normal mucosa tissues; 2. by real-time fluorescent quantitative PCR the expression of IQGAP3 protein was detected by Western blot in colorectal cancer tissues and cell lines, and the construction of stable IQGAP3 overexpression and RNAi cells; 3. by MTT, clone formation, soft agar solid Check and subcutaneous tumor formation experiment, wound healing assay, Transwell assay, to study the effect of IQGAP3 on proliferation and migration of tumor cells; 4. by GSEA gene enrichment analysis and prediction of biological processes involved in IQGAP3, using the method of flow cytometry and Western blot detection IQGAP3 changes related signal pathway or target gene expression after the interference of IQGAP3 in CRC, the occurrence and migration mechanism. Results 1.IQGAP3 expression in colorectal cancer tissues was higher than that of adjacent normal tissues; the expression of 2.IQGAP3 in colorectal cancer cell lines than that in normal intestinal mucosa cells; overexpression of 3.IQGAP3 can significantly promote the proliferation and migration of CRC cells, and IQGAP3 interference can significantly inhibit the proliferation and migration of CRC cells; the expression of 4.IQGAP3 activity and its downstream target genes. The effect of KRAS signaling node The expression level of 1.IQGAP3 in CRC tissues was significantly higher than that in normal mucosa pairing; overexpression of 2.IQGAP3 could significantly promote the proliferation and migration of CRC cells, and IQGAP3 interference can significantly inhibit the proliferation and migration of CRC cells through KRAS/AKT signaling pathway; 3.IQGAP3, KRAS/Rac1, CDC42 signaling pathway, and promote the proliferation of the migration of CRC cells.

【學(xué)位授予單位】：南方醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R735.34

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 Xiao-Lan Li;Jianbiao Zhou;Zhi-Rong Chen;Wee-Joo Chng;;p53 mutations in colorectal cancer-molecular pathogenesis and pharmacological reactivation[J];World Journal of Gastroenterology;2015年01期

2 龐曉雯;閔婕;劉佳鈺;周菁;張峰;劉理禮;張賀龍;;IQGAP1在非小細(xì)胞肺癌腦轉(zhuǎn)移中的作用[J];現(xiàn)代腫瘤醫(yī)學(xué);2013年03期

3 Carla Guarinos;Cristina Sánchez-Fortún;María Rodríguez-Soler;Cristina Alenda;Artemio Payá;Rodrigo Jover;;Serrated polyposis syndrome:Molecular,pathological and clinical aspects[J];World Journal of Gastroenterology;2012年20期

4 Bijan Moghimi-Dehkordi;Azadeh Safaee;;An overview of colorectal cancer survival rates and prognosis in Asia[J];World Journal of Gastrointestinal Oncology;2012年04期

5 劉志輝;宋明旭;周�？�;李莉華;;IQGAP1通過(guò)mTOR信號(hào)通路促進(jìn)肝癌細(xì)胞增殖[J];中國(guó)腫瘤臨床;2011年20期

6 Margaret P.Quinlan;Jeffrey Settleman;;癌癥啟動(dòng)中Kras的特異性功能[J];癌癥;2008年07期

7 盧曉航;;大腸癌發(fā)病機(jī)制研究進(jìn)展[J];現(xiàn)代預(yù)防醫(yī)學(xué);2006年01期

8 王東旭,房殿春,閻曉初,劉為紋,柳鳳軒;抑癌基因APC蛋白在結(jié)直腸粘膜及結(jié)直腸癌中的表達(dá)[J];中華病理學(xué)雜志;1999年06期

9 沈琳;;轉(zhuǎn)移性結(jié)直腸癌靶向治療個(gè)體化選擇[J];腫瘤研究與臨床;2008年08期

10 丁叔波;朱遠(yuǎn);;Kras基因與結(jié)直腸癌[J];浙江臨床醫(yī)學(xué);2009年11期

，

本文編號(hào)：1556222