esVR-2 在食管癌细胞中表达及其调控区UTR 活性的研究
陈涛1，索彩霞1，吴智勇1,3，廖连娣2，谢剑君1，吴健谊1，姚晓冬3，
邹海鹰1，王少洪4，薛玉洁2，许丽艳2，李恩民1

基金项目：中国高等教育博士点科研基金项目资助（20104402110003）
作者简介：陈涛（1984 年-），男，硕士研究生，食管癌分子生物学专业
通信联系人：李恩民（1963 年-），男，教授，食管癌分子生物学. E-mail: nmli@stu.edu.cn
（1. 汕头大学医学院生物化学与分子生物学教研室，汕头，515041；2. 汕头大学医学院肿瘤病理研究室，汕头，515041；3. 汕头市中心医院肿瘤科，汕头，515031；4. 汕头市中心医院病理科，，汕头，515041）
摘要：目的 检测内源性可溶性血管内皮生长因子受体2（endogenous soluble vascular endothelial growth factor receptor-2，esVR-2）在多种食管癌细胞中的表达情况，克隆esVR-2 基因的3′-UTR，探讨esVR-2基因3′UTR 调控报告基因的活性。 方法 采用逆转录聚合酶链反应法（RT-PCR）及定量逆转录聚合酶链反应法(qRT-PCR)，检测esVR-2 在多种食管癌细胞中的mRNA 的表达水平；采用3′RACE 法克隆esVR-2 的3′UTR 区；采用双荧光素酶报告基因分析系统，检测esVR-2 基因3′UTR 区调控报告基因表达的能力。 结果 1）14 种食管鳞癌细胞系中的9 种，esVR-2 的mRNA 的表达水平明显低，占64.3﹪；2）两种食管腺癌细胞系SEG1 和SKTG4，esVR-2 的mRNA 的表达水平，前者明显低，后者明显高；3）5 种永生化食管上皮细胞，esVR-2 的mRNA 的表达水平均明显低；4）与转染空载体pGL-C 的食管癌细胞相比，转染pGL-C3′UTR的食管癌细胞的报告基因荧光素酶活性发生显著变化。结论 1）永生化食管上皮细胞、多数食管鳞癌细胞和部分食管腺癌细胞，esVR-2 的mRNA 的表达水平明显低；2）esVR-2 基因的3′UTR 具有调控报告基因表达的活性。
关键词：内源性可溶性血管内皮生长因子受体2（esVR-2）；食管癌；调控区3′UTR
中图分类号：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)