rna干扰技术对食管癌细胞survivin基因表达的抑制作用及其促凋亡研究-inhibitory effect of rna interference technology on survivin gene expression and its apoptosis promotion in esophageal cancer cells.docxVIP

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rna干扰技术对食管癌细胞survivin基因表达的抑制作用及其促凋亡研究-inhibitory effect of rna interference technology on survivin gene expression and its apoptosis promotion in esophageal cancer cells.docx

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rna干扰技术对食管癌细胞survivin基因表达的抑制作用及其促凋亡研究-inhibitory effect of rna interference technology on survivin gene expression and its apoptosis promotion in esophageal cancer cells

between September 2005 and December 2005 at the Tumor Hospital of Shoutou University Medical College. 31 males and 7 females were between 41 to 78 years old, the median age was58. None of the patients had received preoperative radiotherapy and/or chemotherapy. Dewaxed paraffin sections were immunostained by the Envision method. A modified protocol based on the manufacturer’s instructions was used. Bright field images of immunostained tissues under 400-fold magnification were captured using a Nikon microscope.Results: Survivin expression was predominantly observed in the cytoplasm and nuclear of the tumor cells. The positive rate was 63.16% (24/38). Survivin expression level in the patients with different ages and histopathological grades showed obvious difference. But there were no obvious difference with different sexes and gradings.Conclusions: These results suggested that Survivin expression might be one of the important prognostic indicators for esophageal carcinoma patients, and Survivin might be identified as a potential target in esophageal carcinoma.Keywords: Esophageal carcinoma；Survivin；ImmunohistochemistryPART II Inhibition Effects of RNA Interference Expressing Vector on Survivin GeneObjective: To inhibit the expression of endogenous survivin gene and induce the apoptosis in human esophageal cancer cell line EC-109, small interfering RNA (siRNA) targeting survivin gene was applied.Methods: The recombinant plasmid pSRIEN/S was constructed and transfected into EC-109 cells. The transfection efficiency was determined using fluorescence microscopy. The mRNA silence of survivin gene was detected by semi-quantitative RT-PCR and the expression level of survivin protein was determined by Western blot and Immunofluorescence staining. Theapoptotic rate was observed by the flow cytometry and Genomic DNA Extract Kit.Results: Fluorescence microscopy showed that the transfection efficiency was about 46.7%. Compared with survivin mRNA expression in the contro