Medical Examination Center, The First Affiliated Hospital of Henan University of TCM, Zhengzhou, People’s Republic of China.
Diagn Pathol. 2013 Aug 5;8:132. doi: 10.1186/1746-1596-8-132.
STAT is the backward position of cytokine and growth factor receptors in the nucleus, STAT dimers could bind to DNA and induce transcription of specific target genes. Several lines of evidence support the important roles of STAT, especially STAT5, in carcinogenesis. The overexpression of STAT 5 is related to the differentiation and apoptosis of tumor cells. However, the role of STAT5 in esophageal squamous cell carcinoma remains unclear.
The siRNA vectors aiming to STAT5 gene were constructed. STAT5 siRNA was transfected into Eca-109 cells by Lipofectamine™2000. Expression of STAT5、Bcl-2 and Cyclin D1 were analyzed by Western blot and RT-PCR. Eca-109 cells proliferation was determined by MTT. Eca-109 cell cycle and apoptosis were detected by the flow cytometry. Boyden chamber was used to evaluate the invasion and metastasis capabilities of Eca-109 cells.
The double strands oligonucleotide of siRNA aiming to STAT5 was successfully cloned into the pRNAT-U6.1 vector, and the target sequence coincided with the design. RT-PCR and Western blotting detection demonstrated that the expression levels of STAT5、Bcl-2 and Cyclin D1 gene were obviously decreased in Eca-109 cells transfected with STAT5 siRNA. STAT5 siRNA could suppress the proliferation of Eca-109 cells. The proportion of S and G2/M period frequency was significantly decreased (p<0.05). The proportion of G0/G1 period frequency was significantly increased (p<0.05). The average amount of cells penetrating Matrigel was significantly decreased (p<0.05).
STAT5 silenced by siRNA could induce the apoptosis and suppress the proliferation、invasion and metastasis of esophageal carcinoma cell line Eca-109, which indicated STAT5 might be a novel therapeutic strategy for the human ESCC.
The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1351913072103000.
STAT 是细胞因子和生长因子受体在核内的反向位置，STAT 二聚体可以与 DNA 结合并诱导特定靶基因的转录。有几条证据支持 STAT，尤其是 STAT5，在肿瘤发生中的重要作用。STAT5 的过度表达与肿瘤细胞的分化和凋亡有关。然而，STAT5 在食管鳞状细胞癌中的作用尚不清楚。
构建针对 STAT5 基因的 siRNA 载体。用 LipofectamineTM2000 将 STAT5 siRNA 转染至 Eca-109 细胞。用 Western blot 和 RT-PCR 分析 STAT5、Bcl-2 和 Cyclin D1 的表达。用 MTT 测定 Eca-109 细胞的增殖。用流式细胞术检测 Eca-109 细胞的细胞周期和凋亡。用 Boyden 室评估 Eca-109 细胞的侵袭和转移能力。
成功将靶向 STAT5 的 siRNA 双链寡核苷酸克隆到 pRNAT-U6.1 载体中，靶序列与设计相符。RT-PCR 和 Western blot 检测显示，转染 STAT5 siRNA 的 Eca-109 细胞中 STAT5、Bcl-2 和 Cyclin D1 基因的表达水平明显降低。STAT5 siRNA 可抑制 Eca-109 细胞的增殖。S 和 G2/M 期频率的比例明显降低（p<0.05）。G0/G1 期频率的比例明显增加（p<0.05）。穿过 Matrigel 的细胞平均数量明显减少（p<0.05）。
siRNA 沉默 STAT5 可诱导食管癌细胞系 Eca-109 凋亡，并抑制其增殖、侵袭和转移，表明 STAT5 可能成为人类 ESCC 的一种新的治疗策略。
