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      當前位置:首頁 > 智慧健康列表 > 組織工程研究 > 詳細信息
      esVR-2 在食管癌細胞中表達及其調控區UTR 活性的研究
      來源: | 作者: | 發布時間:2014-4-15 16:17:32

       esVR-2 在食管癌細胞中表達及其調控區UTR 活性的研究
      陳濤1,索彩霞1,吳智勇1,3,廖連娣2,謝劍君1,吳健誼1,姚曉冬3,
      鄒海鷹1,王少洪4,薛玉潔2,許麗艷2,李恩民1

      基金項目:中國高等教育博士點科研基金項目資助(20104402110003)
      作者簡介:陳濤(1984 年-),男,碩士研究生,食管癌分子生物學專業
      通信聯系人:李恩民(1963 年-),男,教授,食管癌分子生物學. E-mail: nmli@stu.edu.cn
      (1. 汕頭大學醫學院生物化學與分子生物學教研室,汕頭,515041;2. 汕頭大學醫學院腫瘤病理研究室,汕頭,515041;3. 汕頭市中心醫院腫瘤科,汕頭,515031;4. 汕頭市中心醫院病理科,,汕頭,515041)
      摘要:目的 檢測內源性可溶性血管內皮生長因子受體2(endogenous soluble vascular endothelial growth factor receptor-2,esVR-2)在多種食管癌細胞中的表達情況,克隆esVR-2 基因的3′-UTR,探討esVR-2基因3′UTR 調控報告基因的活性。 方法 采用逆轉錄聚合酶鏈反應法(RT-PCR)及定量逆轉錄聚合酶鏈反應法(qRT-PCR),檢測esVR-2 在多種食管癌細胞中的mRNA 的表達水平;采用3′RACE 法克隆esVR-2 的3′UTR 區;采用雙熒光素酶報告基因分析系統,檢測esVR-2 基因3′UTR 區調控報告基因表達的能力。 結果 1)14 種食管鱗癌細胞系中的9 種,esVR-2 的mRNA 的表達水平明顯低,占64.3﹪;2)兩種食管腺癌細胞系SEG1 和SKTG4,esVR-2 的mRNA 的表達水平,前者明顯低,后者明顯高;3)5 種永生化食管上皮細胞,esVR-2 的mRNA 的表達水平均明顯低;4)與轉染空載體pGL-C 的食管癌細胞相比,轉染pGL-C3′UTR的食管癌細胞的報告基因熒光素酶活性發生顯著變化。結論 1)永生化食管上皮細胞、多數食管鱗癌細胞和部分食管腺癌細胞,esVR-2 的mRNA 的表達水平明顯低;2)esVR-2 基因的3′UTR 具有調控報告基因表達的活性。
      關鍵詞:內源性可溶性血管內皮生長因子受體2(esVR-2);食管癌;調控區3′UTR
      中圖分類號:R446.8
      Expression of esVR-2 in Esophageal Squamous Cell 25 Carcinoma Cell Lines and Activity of Its 3′-UTR
      CHEN Tao1, SUO Cai-xia1, WU Zhi-yong1,3, LIAO Lian-di2, XIE Jian-jun1, WU
      Jian-yi1, YAO Xiao-dong3, ZOU Hai-ying1, WANG Shao-hong4, XUE Yu-jie2, XU Li-yan2, LI En-min1
      (1. Department of Biochemistry and Molecular Biology, Shantou University Medical College, 30 Shantou 515041;
      2. Department of oncopathology, Shantou University Medical College, Shantou 515041;3. Department of Surgical Oncology, The Shantou Central Hospital, Shantou 515031;4. Department of Pathology, The Shantou Central Hospital, Shantou 515031)
      Abstract: Purpose: The investigation is to detect the expression of endogenous soluble vascular endothelial growth factor receptor2 (esVR2) in esophageal squamous cell carcinoma (ESCC), to clone esVR2 3′UTR and to explore the mechanism of regulation. Methods: Detect the mRNA expression level of esVR2 used Reverse Transcription-Polymerase Chain Reaction (RT-PCR) and quantified Reverse Transcription-Polymerase Chain Reaction (qRT-PCR); cloned 3′UTR of esVR2 use 3′RACE(Rapid Amplification of cDNA Ends); used Double Luciferase Report Gene 40 to detect the regulation of esVR2 3′UTR. Results: Our results showed that: 1) In 14 samples of ESCC, 9 of esVR2 mRNA show a low expression, the percentage comes to 64.3﹪. 2) The level of mRNA of esVR2 in SEG1 appearances a remarkable low expression, while SKTG4 is high. 3)
      The expressions of esVR2 mRNA are all low in 5 Immortalized esophageal epithelial cells. 4) Compared with only transfect pGL-C vector in ESCC, there is an obviously change of the lusiferase activity when transfect pGL-C 45 esVR2. Conclusions: 1) In most esophageal squamous cell carcinoma (ESCC) cells, partial esophageal adenocarcinoma cell carcinoma (EACC) cells and all Immortalized esophageal epithelial cells, the expression of esVR2 mRNA show an obvious low expression;2)There is a regulation Report Gene activity in esVR2 3′UTR.
      Key words: esVR-2(endogenous soluble vascular endothelial growth factor receptor-2);Esophageal Carcinoma; 3′UTR(3′-untranslated region)

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