本文選題：mir-1271 + RTEF-1　； 參考：《鄭州大學》2015年碩士論文

【摘要】：背景與目的胃癌是我國胃腸道腫瘤中常見的惡性腫瘤,具有惡性程度高、易轉移和死亡率高的特點。因此,深入研究胃癌發(fā)病的機制從而尋找到新的早期診斷和有效的治療方法,是擺放在我國醫(yī)療工作者面前的重要問題。胃癌發(fā)病率隨患者年齡增加而升高,其發(fā)病高峰年齡為60歲左右。較之青年胃癌,老年胃癌患者表現出組織學分化程度高、生物學侵襲性弱、腫瘤惡性程度低的臨床特點。老年胃癌與青年胃癌在生物學行為上的這種差別提示兩組胃癌在各自的發(fā)病及發(fā)展過程中存在著不同的分子調節(jié)機制。但是,目前尚不清楚是什么分子機制導致了老年胃癌具有相對低危的生物學特性。因此,研究老年和青年胃癌間存在的不同分子調節(jié)機制,有利于我們進一步深入了解胃癌發(fā)生、發(fā)展的過程,對于胃癌的個體化預防和治療具有重要的理論指導意義。惡性腫瘤普遍具有高度異質性的特點,既往研究結果顯示:老年胃癌是胃癌患者中的一個主要患者群體,其臨床病理特征不同于青年胃癌,反映在:女性發(fā)病率顯著低、組織學分化程度高、生物學侵襲性弱、腫瘤惡性度低等。且老年胃癌遺傳傾向低于青年胃癌,表現為有胃癌家族史的患者明顯較青年胃癌組低。分子學水平上,既往研究也發(fā)現老年胃癌及青年胃癌患者間的基因組譜差異,另外介導細胞黏附和維持細胞骨架的E-鈣黏素和β-Catein表達水平和標志著DNA錯配修復功能缺陷的微衛(wèi)星不穩(wěn)定性(MSI)等也在兩組胃癌間存在著明顯的差異。但是,前期的研究多止步于觀察到上述差異,而缺少對其內在精確分子機制的研究。RTEF-1(Related Transcriptional Enhancer Factor-1,轉錄增強因子-1相關基因)來自轉錄增強子家族(Transcriptional Enhancer Factor,TEF)。1996年Stewart等人從心肌細胞中首次成功克隆并編碼,隨后其家族的其他成員被不斷發(fā)現。目前已知該家族成員包括四個:TEF-1,RTEF-1,DTEF-1和ETF,該家族擁有一個共同的高度保守的DNA結合域,可以結合到基因啟動子M-CAT原件上發(fā)揮其調節(jié)作用。RTEF-1作為轉錄增強子家族重要的組成部分,既往的研究多集中于其在內皮細胞和血管生成中的調節(jié)作用。Che P等人既往的研究結果證實了Mir-125a-5p通過下調RTEF-1可以導致老年小鼠血管生成障礙。鑒于血管生成和腫瘤的密切關系,近來RTEF-1在腫瘤形成中的作用同樣引起了關注。我們提出假設:RTEF-1及相關調控因子在老年胃癌及青年胃癌中的表達是否存在差異,這種差異是否與老年胃癌和青年胃癌間的組織學分化程度、生物學侵襲性及腫瘤惡性程度的不同有關。目的:探討RTEF-1及其上游調控基因在老年胃癌及青年胃癌中的表達是否存在差異及其意義。材料與方法1.研究對象及標本采集:標本采集于鄭州大學第一附屬醫(yī)院胃腸外科2014年7月至2015年1月期間進行胃癌根治術的患者。納入標準:術前胃鏡病理提示為胃惡性腫瘤,計算機斷層掃描(computed tomography,CT)等檢查進行術前分期,提示患者可以行標準胃癌根治術,術后病理證實為胃癌中的低分化腺癌,術前未進行化療,放療及中西醫(yī)結合等抗腫瘤治療。本實驗經我院道德倫理委員會的批準,而且經過患者知情同意并簽署知情同意書。2.自Microarray數據結果中分析老年胃癌和青年胃癌中存在差異表達的mi RNAs;發(fā)現在老年胃癌及青年胃癌中表達存在明顯差異的mi RNA,RT-q PCR驗證;3.分別采用RT-q PCR、Western Blot技術檢測RTEF-1在老年胃癌及青年胃癌原代細胞中的m RNA蛋白水平表達并進行比較。4.所有實驗結果數據均采用SPSS17.0軟件進行分析。以P0.05作為差異有統(tǒng)計學意義的檢驗標準。結果1.mir-1271在老年胃癌和青年胃癌中均含量豐富且變化相對明顯;2.老年胃癌細胞中RTEF-1蛋白表達較青年胃癌減少;3.老年胃癌細胞中mir-1271表達是青年胃癌的3.2倍;結論老年胃癌及青年胃癌中mir-1271及RTEF-1存在差異性表達,這種差異可能是導致老年胃癌及青年胃癌在組織學分化程度、生物學侵襲性及腫瘤惡性程度存在差異的重要分子機制。
[Abstract]:Background and objective gastric cancer is a common malignant tumor of the gastrointestinal tract in China. It has the characteristics of high malignancy, easy transfer and high mortality. Therefore, it is an important problem placed in front of the medical workers in our country to study the pathogenesis of gastric cancer so as to find a new early diagnosis and effective treatment method. The age of the patients increased and the peak age was about 60 years old. Compared with the young gastric cancer, the elderly patients with gastric cancer showed high histological differentiation, weak biological invasiveness and low malignancy. This difference in biological behavior between the aged and the young gastric cancer suggests that the two groups of gastric cancer are in their respective pathogenesis and hair. There are different molecular mechanisms in the process. However, it is not clear what molecular mechanism leads to the relatively low risk of biological characteristics in elderly gastric cancer. Therefore, the study of different molecular mechanisms in the elderly and young gastric cancer is helpful for us to further understand the occurrence and development of gastric cancer and to the stomach. The individualized prevention and treatment of cancer have important theoretical guiding significance. The malignant tumor has the characteristics of high heterogeneity. The previous research results show that the elderly gastric cancer is a major group of patients with gastric cancer, whose clinicopathological features are different from those of young gastric cancer, which are reflected in the low incidence of female and high degree of histological differentiation. The genetic tendency of the old gastric cancer is lower than that of the young gastric cancer, and the patients with the family history of gastric cancer are obviously lower than those of the young gastric cancer group. The expression level of E- calcarin and beta -Catein and the microsatellite instability (MSI) indicating the functional defect of DNA mismatch repair (MSI) were also significantly different between the two groups of gastric cancers. However, the previous study stopped to observe the above differences, but there was a lack of.RTEF-1 (Related Transcriptional Enhancer) for its intrinsic precise molecular mechanism. Factor-1, -1 related genes from the transcription enhancer family (Transcriptional Enhancer Factor, TEF).1996 Stewart et al. Stewart et al. The first successful cloning and encoding from the cardiac myocytes, and then the other members of the family are constantly discovered. Now the family members are known to include four: TEF-1, RTEF-1, DTEF-1 and ETF, the family embraced There is a common highly conservative DNA binding domain that can be combined with the gene promoter M-CAT to play its regulatory role.RTEF-1 as an important component of the transcription enhancer family. Previous studies have concentrated on its regulatory role in endothelial cells and angiogenesis, and the previous results of.Che P, et al., confirmed that Mir-125a-5p Down regulation of RTEF-1 can lead to angiogenic disorders in old mice. In view of the close relationship between angiogenesis and tumor, the recent role of RTEF-1 in tumor formation is also concerned. We hypothesized: whether there is a difference in the expression of RTEF-1 and related regulatory factors in old and young gastric cancer, whether this difference is with the elderly Histological differentiation, biological invasiveness and tumor malignancy of gastric cancer and young gastric cancer. Objective: To investigate whether the expression of RTEF-1 and its upstream regulatory genes in old gastric cancer and young gastric cancer are different and its significance. Materials and methods 1. research objects and specimen collection: specimen collection at the first of Zhengzhou University Patients undergoing radical gastrectomy for gastric cancer from July 2014 to January 2015 of the affiliated hospital were included in the criteria: preoperative gastroscopic pathological hints for gastric malignant tumor, computed tomography (computed tomography, CT) and other examinations for preoperative staging, suggesting that patients can be treated with standard radical gastrectomy, and the postoperative pathology proved to be a low score in gastric cancer. Adenocarcinoma, chemotherapy, radiotherapy and combination of traditional Chinese and Western medicine were not performed before operation. This experiment was approved by the ethics committee of our hospital, and after the informed consent of the patients and the signing of the informed consent form.2. from the Microarray data, the differential expression of MI RNAs in old gastric cancer and young gastric cancer was analyzed, and it was found in old gastric cancer and Mi RNA, RT-q PCR, and RT-q PCR, Western Blot technology were used to detect the expression of M RNA protein in old gastric cancer and young gastric cancer cells by RT-q PCR and Western Blot, respectively, and the results of all experimental data were analyzed. Results 1.mir-1271 was rich in both old and young gastric cancer. The expression of RTEF-1 protein in 2. elderly gastric cancer cells was less than that in young gastric cancer, and the expression of mir-1271 in 3. old gastric cancer cells was 3.2 times as high as that of young gastric cancer. Conclusion there were differences in mir-1271 and RTEF-1 in old gastric cancer and young gastric cancer. Sexual expression, which may be an important molecular mechanism that leads to differences in histological differentiation, biological invasiveness and malignancy of old and young gastric cancer.
【學位授予單位】：鄭州大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R735.2

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