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RNA 干扰介导的 UBCH10 基因沉默抑制结直肠癌细胞的体内外生长。

RNA interference-mediated silencing of UBCH10 gene inhibits colorectal cancer cell growth in vitro and in vivo.

机构信息

Institute of Biochemistry and Molecular Biology, Shandong University School of Medicine, Wenhua Xi Road, Jinan, 250012 China.

出版信息

Clin Exp Pharmacol Physiol. 2010 May;37(5-6):525-9. doi: 10.1111/j.1440-1681.2009.05348.x.

DOI:10.1111/j.1440-1681.2009.05348.x
PMID:20529090
Abstract
  1. UbcH10 is the cancer-related E2 ubiquitin-conjugating enzyme, and its overexpression has been demonstrated in a variety of malignancies. The aim of the present study is to silence UbcH10 gene by RNA interference (RNAi) and to observe its inhibitory effect on the colorectal cancer cell growth in vitro and in vivo. 2. We constructed the expression vector pGPU6/GFP/Neo/UbcH10-RNAi (pUbcH10-RNAi), which contained a UbcH10 short hairpin RNA expression cassette. Then the UbcH10 gene silencing cell lines LoVo/UbcH10-RNAi and HT-29/UbcH10-RNAi were established. Reverse transcription-polymerase chain reaction and western blot analysis were used to evaluate the expression of the UbcH10 gene. Cell Counting Kit-8 was used to assess properties of tumour cell growth in vitro. Flow cytometry was used to detect the effect of pUbcH10-RNAi on the cell cycle of colorectal cancer cells. Furthermore, the anti-tumour effects of pUbcH10-RNAi were evaluated in vivo in a nude mouse xenografts model. 3. Results demonstrated that UbcH10 gene expression was significantly decreased in pUbcH10-RNAi treated cells. Colorectal cancer cells growth was markedly suppressed in the pUbcH10-RNAi group compared with control conditions and colorectal cancer cells were arrested in the G2-M phase. In vivo, the downregulation of UbcH10 gene expression by pUbcH10-RNAi also inhibited tumour growth in a nude mice xenograft model. 4. Our study suggests that RNA interference-mediated silencing of UbcH10 gene has anti-tumour activity on colorectal cancer and might have therapeutic potential for the treatment of colorectal cancer.
摘要
  1. UbcH10 是一种与癌症相关的 E2 泛素缀合酶,其在多种恶性肿瘤中过表达。本研究旨在通过 RNA 干扰(RNAi)沉默 UbcH10 基因,并观察其对体外和体内结直肠癌细胞生长的抑制作用。

  2. 我们构建了表达载体 pGPU6/GFP/Neo/UbcH10-RNAi(pUbcH10-RNAi),其中包含 UbcH10 短发夹 RNA 表达盒。然后建立了 UbcH10 基因沉默细胞系 LoVo/UbcH10-RNAi 和 HT-29/UbcH10-RNAi。逆转录-聚合酶链反应和 Western blot 分析用于评估 UbcH10 基因的表达。细胞计数试剂盒-8 用于评估体外肿瘤细胞生长特性。流式细胞术用于检测 pUbcH10-RNAi 对结直肠癌细胞周期的影响。此外,在裸鼠异种移植模型中评估了 pUbcH10-RNAi 的抗肿瘤作用。

  3. 结果表明,pUbcH10-RNAi 处理的细胞中 UbcH10 基因表达明显降低。与对照条件相比,pUbcH10-RNAi 组结直肠癌细胞生长明显受到抑制,细胞停滞在 G2-M 期。在体内,pUbcH10-RNAi 下调 UbcH10 基因表达也抑制了裸鼠异种移植模型中的肿瘤生长。

  4. 我们的研究表明,RNAi 介导的 UbcH10 基因沉默对结直肠癌细胞具有抗肿瘤活性,可能为结直肠癌的治疗提供潜在的治疗方法。

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RNA interference-mediated silencing of UBCH10 gene inhibits colorectal cancer cell growth in vitro and in vivo.RNA 干扰介导的 UBCH10 基因沉默抑制结直肠癌细胞的体内外生长。
Clin Exp Pharmacol Physiol. 2010 May;37(5-6):525-9. doi: 10.1111/j.1440-1681.2009.05348.x.
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Association of clinicopathological features with UbcH10 expression in colorectal cancer.结直肠癌中 UbcH10 表达与临床病理特征的关系。
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RNAi-mediated silencing of VEGF-C inhibits non-small cell lung cancer progression by simultaneously down-regulating the CXCR4, CCR7, VEGFR-2 and VEGFR-3-dependent axes-induced ERK, p38 and AKT signalling pathways.RNAi 介导的 VEGF-C 沉默通过同时下调 CXCR4、CCR7、VEGFR-2 和 VEGFR-3 依赖性轴诱导的 ERK、p38 和 AKT 信号通路抑制非小细胞肺癌的进展。
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