本文選題：食管癌 + microRNA��； 參考：《第四軍醫(yī)大學》2015年碩士論文

【摘要】：【背景】食管癌是世界上第八大常見惡性腫瘤,第六大因癌癥導致死亡的原因。目前全世界有45萬多食管癌患者,并且其發(fā)病率仍在上升。食管癌診斷越早,預(yù)后越好。然而,由于缺乏有效的早期診斷手段,大部分患者首診時已屆進展期,其總的五年生存率僅為15%~25%,因此有必要探索有應(yīng)用價值的食管癌分子標志物和靶分子,從而為食管癌的早期診斷和治療提供線索。micro RNAs(mi RNAs)是一類在進化過程中高度保守的長度在18~25個核苷酸的單鏈非編碼小RNA,主要通過抑制靶基因的翻譯影響人體的各種生理和病理過程。前期我們通過mi RNA芯片檢測了25例食管鱗癌及其癌旁組織的差異表達分子,率先發(fā)現(xiàn)并報道了一個食管癌發(fā)生相關(guān)的mi RNA分子群。其中,mi R-483在食管癌組織中高表達,但其對食管癌細胞惡性表型的影響及分子機制尚未闡明,本課題將對此展開研究。【目的】1.檢測mi R-483-3p在食管鱗癌細胞系及正常食管鱗狀上皮細胞系中的表達;2.構(gòu)建mi R-483-3p過表達和低表達的食管癌細胞模型,通過體內(nèi)外實驗研究mi R-483-3p對食管癌生物學行為的影響;3.探討mi R-483-3p影響食管癌發(fā)生的分子機制�！痉椒ā�1.q PCR檢測mi R-483-3p在食管鱗癌細胞系及正常食管鱗狀上皮細胞系中的表達;2.構(gòu)建mi R-483-3p的慢病毒表達載體及空載體,轉(zhuǎn)染食管癌細胞,建立過表達mi R-483-3p的細胞模型及相應(yīng)對照細胞模型;3.合成mi R-483-3p inhibitor及inhibitor NC,轉(zhuǎn)染食管癌細胞,建立低表達mi R-483-3p的細胞模型及相應(yīng)對照細胞模型;4.MTT實驗研究mi R-483-3p對食管癌細胞增殖的影響;5.Transwell實驗觀察mi R-483-3p對食管癌細胞遷移的影響;6.流式細胞儀檢測mi R-483-3p對食管鱗癌細胞周期的影響;7.流式細胞儀檢測mi R-483-3p對食管癌細胞凋亡的影響;8.MTT法體外藥物敏感性實驗檢測mi R-483-3p對食管癌細胞藥物敏感性的影響;9.裸鼠成瘤實驗觀察mi R-483-3p對食管癌細胞體內(nèi)成瘤能力的影響;10.通過生物信息學及全基因組表達譜芯片聯(lián)合預(yù)測的方法篩選mi R-483-3p的靶基因;11.通過q PCR、Western blot及雙熒光素酶報告基因?qū)嶒烌炞Cmi R-483-3p的靶基因�！窘Y(jié)果】1.與正常食管鱗狀上皮細胞系Het-1A相比,mi R-483-3p在食管鱗癌細胞系EC109、EC9706及TE-1中表達升高。2.成功建立了過表達和低表達mi R-483-3p的食管癌細胞模型。3.上調(diào)mi R-483-3p可促進食管癌細胞的增殖、遷移,促進細胞周期由G1期向G2期轉(zhuǎn)化,減少ADR誘導的細胞凋亡,減弱細胞對化療藥物的敏感性,增強食管癌細胞的體內(nèi)成瘤能力。4.下調(diào)mi R-483-3p可抑制食管癌細胞的增殖、遷移,導致細胞周期G1期阻滯,增加ADR誘導的細胞凋亡,增強細胞對化療藥物的敏感性。5.通過生物信息學和全基因組表達譜芯片聯(lián)合預(yù)測,得到7個mi R-483-3p的靶基因:EI24、KLHL12、GRSF1、MKRN1、LEPROTL1、CREBL2、MAPKAPK2。6.上調(diào)食管癌細胞中mi R-483-3p的表達對以上7個靶基因的m RNA表達水平無影響。7.上調(diào)食管癌細胞中mi R-483-3p的表達可導致抑癌基因EI24的表達下降。【結(jié)論】1.與正常食管鱗狀上皮細胞相比,mi R-483-3p在食管鱗癌細胞系中表達升高。2.上調(diào)mi R-483-3p表達可通過調(diào)節(jié)細胞周期促進食管癌細胞的增殖、遷移,增強細胞對化療藥物的耐藥性;下調(diào)mi R-483-3p則起到相反的效果。3.mi R-483-3p可能通過調(diào)節(jié)EI24等靶基因的表達影響食管癌的發(fā)生發(fā)展。
[Abstract]:[background] esophageal cancer is the 8th most common malignant tumor in the world. The sixth major cause of death is cancer. There are more than 450 thousand esophageal cancer patients worldwide, and the incidence is still rising. The earlier the diagnosis of esophageal cancer is, the better the prognosis. However, because of the lack of effective early diagnosis, most of the patients have progressing in the first visit. The total five year survival rate is only 15%~25%, so it is necessary to explore the useful molecular markers and target molecules of esophageal cancer, thus providing clues to the early diagnosis and treatment of esophageal cancer,.Micro RNAs (MI RNAs) is a class of highly conserved single strand non coded small RNA, which is highly conservative in the course of evolution, with the length of 18~25 nuclear acid, mainly through the suppression of the target. The translation of the gene affects the physiological and pathological processes of the human body. In the earlier period, we detected the differential expression molecules of 25 cases of esophageal squamous cell carcinoma and its adjacent tissues by Mi RNA chip, and first discovered and reported a mi RNA group related to the occurrence of esophageal cancer. Among them, Mi R-483 was highly expressed in the esophageal cancer tissue, but it was bad for the cancer of the esophagus. The effects and molecular mechanisms of sexual phenotype have not been elucidated. [Objective] [Objective] 1. to detect the expression of MI R-483-3p in esophageal squamous cell carcinoma cell lines and normal esophageal squamous cell lines; 2. to construct mi R-483-3p overexpressed and low expressed esophageal cancer cell models, and to study mi R-483-3p for esophageal cancer in vitro and in vivo The influence of physical behavior; 3. to explore the molecular mechanism of MI R-483-3p affecting the occurrence of esophageal cancer. [Methods] 1.q PCR was used to detect the expression of MI R-483-3p in esophageal squamous cell carcinoma cell lines and normal esophageal squamous cell lines; 2. to construct the MI R-483-3p Lentivirus Expression Vector and unloaded body, transfection of esophageal cancer cells, and to establish a fine expression of MI R-483-3p. Cell model and corresponding control cell model; 3. mi R-483-3p inhibitor and inhibitor NC were synthesized to transfect esophageal cancer cells to establish a cell model with low expression of MI R-483-3p and a corresponding control cell model. 4.MTT experiment studied the effect of MI R-483-3p on the proliferation of esophageal cancer cells; 5.Transwell experiment observed the migration of the esophagus cancer cells by MI. The effect of MI R-483-3p on the cell cycle of esophageal squamous cell carcinoma was detected by 6. flow cytometry, and the effect of MI R-483-3p on the apoptosis of esophageal cancer cells was detected by 7. flow cytometry; the effect of MI R-483-3p on the drug sensitivity of esophageal cancer cells was detected by 8.MTT assay in vitro; and 9. in nude mice to observe the effect of MI R-483-3p on esophageal cancer cells. The effect of tumor forming ability; 10. screening target genes of MI R-483-3p by bioinformatics and whole genome expression chip combined prediction; 11. test the target gene of MI R-483-3p by Q PCR, Western blot and double luciferase reporter gene test. [results] 1., MI R-483-3p is compared with the normal esophageal squamous cell line Het-1A. The expression of esophageal squamous cell carcinoma cell lines EC109, EC9706 and TE-1 elevated.2. successfully established the overexpression and low expression of MI R-483-3p in the esophageal cancer cell model.3. to promote the proliferation and migration of the esophageal cancer cells and promote the transformation of the cell cycle from G1 to G2 phase, reducing the apoptosis of ADR induced cells and reducing the sensitivity of cell to chemotherapeutic drugs. Sex, enhancing the tumorigenicity of esophageal cancer cells in vivo.4. down regulation of MI R-483-3p can inhibit the proliferation, migration, cell cycle G1 block, increase ADR induced apoptosis and enhance cell sensitivity to chemotherapeutic drugs,.5. through bioinformatics and total gene group expression chip combined prediction to obtain 7 mi R-483-3p Target genes: EI24, KLHL12, GRSF1, MKRN1, LEPROTL1, CREBL2, MAPKAPK2.6. up-regulated the expression of MI R-483-3p in esophageal cancer cells. The expression of M RNA on the above 7 target genes has no effect on the expression of.7. up esophageal cancer cells. [Conclusion] 1. compared with normal esophageal squamous cells. I R-483-3p increased the expression of.2. in esophageal squamous cell carcinoma cell line and increased the expression of MI R-483-3p by regulating cell cycle to promote the proliferation, migration, and drug resistance of esophageal cancer cells. The downregulation of MI R-483-3p may play the opposite effect,.3.mi R-483-3p may affect the development of esophageal cancer by regulating the expression of EI24 and other target genes. The development of life.
【學位授予單位】：第四軍醫(yī)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R735.1

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