本文選題：LATS1 + 食管鱗狀細胞癌�。� 參考：《重慶醫(yī)科大學》2017年碩士論文

【摘要】：背景:大腫瘤抑制基因1(LATS1)是Hippo信號通路中的一個關(guān)鍵抑癌基因。有研究表明在多種腫瘤中LATS1基因存在啟動子甲基化,并且證實與其發(fā)生發(fā)展相關(guān)聯(lián),但是在食管鱗狀細胞癌中LATS1的功能及與甲基化狀態(tài)的關(guān)系尚未有研究。目的:了解LATS1基因表達與LATS1甲基化狀態(tài)的聯(lián)系,及LATS1在食管鱗狀細胞癌中的功能,并探討LATS1作為食管鱗癌基因靶向治療的可能性。方法:1.通過qRT-PCR、免疫組化檢測LATS1蛋白及mRNA在食管鱗癌及配對癌旁組織中的表達水平差異,并探討與臨床病理特點及分期的關(guān)系。2.甲基化特異性PCR(MSP)和亞硫酸氫鹽測序法(BGS)檢測食管鱗癌組織與細胞中LATS1基因啟動子Cp G島是否發(fā)生了甲基化。3.慢病毒轉(zhuǎn)染食管鱗癌細胞株TE-10和EC109,穩(wěn)定表達LATS1基因。4.CCK-8和克隆形成實驗檢測LATS1基因?qū)E-10和EC109細胞增殖的影響。5.采用流式細胞術(shù)檢測LATS1基因?qū)E-10和EC109細胞周期及細胞凋亡的影響。6.通過transwell實驗檢測LATS1基因?qū)E-10和EC109細胞遷移和侵襲能力的影響。7.使用SPSS23.0對研究所得數(shù)據(jù)進行統(tǒng)計學分析。結(jié)果:1.LATS1基因在兩種組織中表達水平有明顯差異,癌組織中普遍低表達,而癌旁組織中則廣泛表達。2.在食管鱗癌細胞TE-10及EC109中過表達LATS1基因能抑制細胞增殖。3.在TE-10及EC109細胞中過表達LATS1基因能阻滯細胞周期于G2/M期,同時能誘導細胞凋亡。4.在TE-10及EC109細胞中過表達LATS1基因能減少細胞的遷移和侵襲力。5.在食管鱗狀細胞癌組織及細胞中,LAST1基因啟動子CpG島未發(fā)現(xiàn)甲基化位點。結(jié)論:我們的實驗結(jié)果表明LATS1在食管鱗癌中起到抑癌基因的作用,并且與腫瘤的臨床分期相關(guān),而其下調(diào)與CpG島的高甲基化無關(guān)。在食管鱗癌細胞中過表達LATS1明顯下調(diào)YAP,抑制細胞增殖、細胞遷移和侵襲,同時能夠促進細胞凋亡。因此,LATS1基因值得進一步探索作為食管鱗癌生物標志物的可能用途和未來分子診斷和治療的目標。
[Abstract]:Background: large tumor suppressor gene 1 (LATS1) is a key tumor suppressor gene in Hippo signaling pathway. Some studies have shown that there is promoter methylation of LATS1 gene in many kinds of tumors and has been proved to be associated with its occurrence and progression. However, the function of LATS1 and its relationship with methylation status have not been studied in esophageal squamous cell carcinoma. Objective: to investigate the relationship between LATS1 gene expression and LATS1 methylation, and the function of LATS1 in esophageal squamous cell carcinoma, and to explore the possibility of LATS1 as a target therapy for esophageal squamous cell carcinoma. Method 1: 1. The expression of LATS1 protein and mRNA in esophageal squamous cell carcinoma and paracancerous tissues were detected by using qRT-PCR, and the relationship between the expression of mRNA and clinicopathological features and staging was discussed. Methylation specific LATS1 gene promoter CpG island was detected in esophageal squamous cell carcinoma tissues and cells by methylation specific PCR and bisulfite sequencing. Lentivirus was transfected into esophageal squamous cell carcinoma cell lines TE-10 and EC109, and LATS1 gene was stably expressed. 4. CCK-8 and clone formation assay were used to detect the effect of LATS1 gene on the proliferation of TE-10 and EC109 cells. The effect of LATS1 gene on TE-10 and EC109 cell cycle and apoptosis was detected by flow cytometry. The effect of LATS1 gene on the migration and invasion of TE-10 and EC109 cells was detected by transwell assay. Use SPSS23.0 to carry on statistical analysis to the data obtained from the study. Results: 1. The expression level of LATS1 gene was significantly different between the two tissues. The expression of LATS1 gene was generally low in cancer tissues, but widely expressed in paracancerous tissues. Overexpression of LATS1 gene in esophageal squamous cell TE-10 and EC109 can inhibit cell proliferation. 3. Overexpression of LATS1 gene in TE-10 and EC109 cells could block cell cycle in G 2 / M phase and induce apoptosis. Overexpression of LATS1 gene in TE-10 and EC109 cells can reduce cell migration and invasiveness. No methylation site was found in CpG island of LAST1 gene promoter in esophageal squamous cell carcinoma tissues and cells. Conclusion: our results suggest that LATS1 plays a role as a tumor suppressor gene in esophageal squamous cell carcinoma and is related to the clinical stage of the tumor, but its down-regulation is not associated with hypermethylation of CpG island. Overexpression of LATS1 in esophageal squamous cell carcinoma cells significantly down-regulated Yap, inhibited cell proliferation, cell migration and invasion, and promoted apoptosis. Therefore, the LATS1 gene is worthy of further exploration as a possible biomarker for esophageal squamous cell carcinoma and a target for molecular diagnosis and treatment in the future.
【學位授予單位】：重慶醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R735.1

【參考文獻】

相關(guān)期刊論文 前2條

1 María José Domper Arnal;ángel Ferrández Arenas;ángel Lanas Arbeloa;;Esophageal cancer: Risk factors,screening and endoscopic treatment in Western and Eastern countries[J];World Journal of Gastroenterology;2015年26期

2 Piotr M Wierzbicki;Krystian Adrych;Dorota Kartanowicz;Marcin Stanislawowski;Anna Kowalczyk;Janusz Godlewski;Iwona Skwierz-Bogdanska;Krzysztof Celinski;Tomasz Gach;Jan Kulig;Bartlomiej Korybalski;Zbigniew Kmiec;;Underexpression of LATS1 TSG in colorectal cancer is associated with promoter hypermethylation[J];World Journal of Gastroenterology;2013年27期

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本文編號：1935427