Effect of STAT5 silenced by siRNA on proliferation apoptosis and invasion of esophageal carcinoma cell line Eca-109

Diagnostic Pathology

Guoqiang Zhao⁴  Wenqiao Zang⁴  Hong Xu²  Tao Wang¹  Min Li⁴  Qian Yang³ 

[1] Department of Hemato-tumor, The First Affiliated Hospital of Henan University of TCM, Zhengzhou, People’s Republic of China;Henan Tumor Institute, Zhengzhou, People’s Republic of China;Medical Examination Center, The First Affiliated Hospital of Henan University of TCM, Zhengzhou, People’s Republic of China;College of Basic Medical Sciences, Zhengzhou University, Zhengzhou, People’s Republic of China

关键词: Apoptosis;    Cell cycle;    Proliferation;    siRNA;    STAT5;   

Others  :  805740
DOI  :  10.1186/1746-1596-8-132

 received in 2013-07-06, accepted in 2013-07-23,  发布年份 2013

【 摘 要 】

Background

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.

Methods

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.

Results

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).

Conclusions

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.

Virtual slides

The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1351913072103000 webcite

【 授权许可】

   
2013 Yang et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

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