Study on the expression of CXCR4 gene silenced by CXCR4-siRNA recombinant adenoviral vector in human ovarian cancer cells

LIU Yu; YUAN Rui

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Study on the expression of CXCR4 gene silenced by CXCR4-siRNA recombinant adenoviral vector in human ovarian cancer cells

更新时间：2024-01-17

- Study on the expression of CXCR4 gene silenced by CXCR4-siRNA recombinant adenoviral vector in human ovarian cancer cells
- Journal of Chongqing Medical University Vol. 36, Issue 10, Pages: 1168-1171(2011)
- 作者机构：
  
  重庆医科大学附属第一医院妇产科
- 作者简介：
- 基金信息：
- DOI：
  CLC： R737.31
- Published：2011，
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[1]刘瑜,袁瑞.CXCR4-siRNA重组腺病毒载体沉默人卵巢癌细胞株CXCR4基因表达的研究[J].重庆医科大学学报,2011,36(10):1168-1171.

LIU Yu, YUAN Rui. Study on the expression of CXCR4 gene silenced by CXCR4-siRNA recombinant adenoviral vector in human ovarian cancer cells. [J]. Journal of Chongqing Medical University 36(10):1168-1171(2011)
[1]刘瑜,袁瑞.CXCR4-siRNA重组腺病毒载体沉默人卵巢癌细胞株CXCR4基因表达的研究[J].重庆医科大学学报,2011,36(10):1168-1171. DOI：

LIU Yu, YUAN Rui. Study on the expression of CXCR4 gene silenced by CXCR4-siRNA recombinant adenoviral vector in human ovarian cancer cells. [J]. Journal of Chongqing Medical University 36(10):1168-1171(2011) DOI：

摘要

目的:研究CXCR4-siRNA重组腺病毒载体对人卵巢癌细胞株SKOV3的CXCR4基因表达的影响。方法:PacⅠ酶切CXCR4-siRNA重组腺病毒载体使其线性化

应用HEK293细胞大量包装扩增病毒

收集病毒液

转染人卵巢癌细胞株SKOV3。细胞分为3组:siRNA组

转染空病毒载体的阴性对照组

细胞不做任何处理的空白对照组。通过PCR、Western blot、免疫组化检测CXCR4基因沉默效果。结果:包装扩增后病毒滴度为6×108 PFU/ml

荧光显微镜下可观察到SKOV3细胞中绿色荧光蛋白的大量表达

重组腺病毒能够有效抑制SKOV3细胞中CXCR4的表达。结论:CXCR4-siRNA重组腺病毒载体可高效转染靶细胞和沉默CXCR4基因的表达

为RNA干扰技术应用于卵巢癌的临床基因治疗奠定基础。

Abstract

Objective:To study the effect of the CXCR4-siRNA recombinant adenoviral vector on the expression of CXCR4 gene in SKOV3 cells.Methods:Recombinant adenoviral vector was linearized with PacⅠ.Then it was repeatedly packaged and amplified in HEK293 cells and transfected into SKOV3 cells.The SKOV3 cells were divided into three groups:siRNA group with adenovirus carrying RNA interference

negative group with vacant adenovirus vector

and control group without any infection.The expression level of CXCR4 gene in SKOV3 cells was measured by RT-PCR

Western blot and immunocytochemistry analysis.Results:The viral titers were 6×108 PFU/ml.The green fluorescent protein expressed highly in SKOV3 cells under fluorescent microscope.The CXCR4 gene was greatly inhibited after infection with CXCR4-siRNA recombinant adenoviral vector.Conclusion:The CXCR4-siRNA recombinant adenoviral vector can efficiently transfect targeted cells and silence the expression of CXCR4 gene

which would lay a foundation for the further study on clinical gene therapy of RNAi technology in ovarian cancer.

关键词

CXCR4RNA干扰重组腺病毒载体卵巢癌

Keywords

CXCR4RNA interferencerecombinant adenoviral vectorovarian cancer

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