Zhou Cui-Qi, Liu Shuang, Xue Li-Yan, Wang Yi-Hua, Zhu Hong-Xia, Lu Ning, Xu Ning-Zhi
Laboratory of Cell and Molecular Biology, Cancer Institute, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing 100021, China.
World J Gastroenterol. 2003 Sep;9(9):1900-3. doi: 10.3748/wjg.v9.i9.1900.
To investigate gene expression pattern of human gamma-synuclein gene in human esophageal squamous cell carcinoma (ESCC) by using semi-quantitive reverse transcription polymerase chain reaction (RT-PCR), and to study the role of gamma-synuclein in the development of human ESCC.
Semi-quantitive RT-PCR of 27 pairs of specimens of human ESCC tissues and corresponding normal tissues was used to investigate the expression pattern of gamma-synuclein in ESCC. 9706/gamma-syn cells in which gamma-synuclein was overexpressed were obtained through cloning gamma-synuclein gene by PCR and transfecting it into ESCC 9706 cells, then selecting with G-418 for 14 days. The biological effects of gamma-synuclein were measured and compared between 9706/gamma-syn and 9706/vec cells by cell growth curve and soft agar assay.
RT-PCR showed that gamma-synuclein gene was expressed in all the 27 cases of normal epithelial tissues, while downregulation of gamma-synuclein was observed in 16 out of the 27 cases (59.3 %) of ESCC. There were also 6 cases of ESCC tissues with a high expression level of gamma-synuclein mRNA. In functional analysis we found that over-expression of gamma-synuclein in ESCC 9706 cells could inhibit the growth rate and transformation ability of ESCC 9706 cells.
The low expression level of gamma-synuclein in human ESCC and the biological effects of gamma-synuclein over-expression on ESCC 9706 cells suggest that gamma-synuclein may play a role as a negative regulator in the development of human ESCC.
采用半定量逆转录聚合酶链反应(RT-PCR)研究人γ-突触核蛋白基因在人食管鳞状细胞癌(ESCC)中的基因表达模式,并探讨γ-突触核蛋白在人ESCC发生发展中的作用。
运用半定量RT-PCR检测27对人ESCC组织及其相应正常组织标本,以研究γ-突触核蛋白在ESCC中的表达模式。通过PCR克隆γ-突触核蛋白基因并将其转染至ESCC 9706细胞,经G-418筛选14天,获得γ-突触核蛋白过表达的9706/γ-突触核蛋白细胞。通过细胞生长曲线和软琼脂试验检测并比较9706/γ-突触核蛋白细胞和9706/载体细胞中γ-突触核蛋白的生物学效应。
RT-PCR显示,27例正常上皮组织均表达γ-突触核蛋白基因,而27例ESCC组织中有16例(59.3%)γ-突触核蛋白表达下调。另外,有6例ESCC组织γ-突触核蛋白mRNA表达水平较高。在功能分析中我们发现,ESCC 9706细胞中γ-突触核蛋白过表达可抑制ESCC 9706细胞的生长速率和转化能力。
人ESCC中γ-突触核蛋白表达水平较低,且γ-突触核蛋白过表达对ESCC 9706细胞具有生物学效应,提示γ-突触核蛋白可能在人ESCC发生发展中作为负调节因子发挥作用。