RNA interference-mediated silencing of UBCH10 gene inhibits colorectal cancer cell growth in vitro and in vivo
RNA interference-mediated silencing of UBCH10 gene inhibits colorectal cancer cell growth in vitro and in vivo
复制标题
DOI：
10.1111/j.1440-1681.2009.05348.x
复制
发表时间：
2010-05-01
影响因子：
2.9
通讯作者：
Liu, Xian-Xi
中科院分区：
文献类型：
--
作者：
Chen, Shi-Min;Jiang, Chun-Ying;Liu, Xian-Xi
P>1。UbcH 10是与癌症相关的E2泛素结合酶，其过表达已在多种恶性肿瘤中得到证实。本研究的目的是利用RNA干扰技术沉默UbcH 10基因，观察其对大肠癌细胞体内外生长的抑制作用.我们构建了含有UbcH 10短发夹RNA表达盒的表达载体pGPU 6/GFP/Neo/UbcH 10-RNAi（pUbcH 10-RNAi）。建立了UbcH 10基因沉默细胞系LoVo/UbcH 10-RNAi和HT-29/UbcH 10-RNAi。逆转录-聚合酶链反应（RT-PCR）和蛋白质印迹（Western blot）分析UbcH 10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的特性。流式细胞仪检测pUbcH 10-RNAi对大肠癌细胞周期的影响。在裸鼠移植瘤模型中评价pUbcH 10-RNAi的体内抗肿瘤作用.结果表明，经pUbcH 10-RNAi处理的细胞中UbcH 10基因的表达显著降低。与对照条件相比，pUbcH 10-RNAi组中结肠直肠癌细胞的生长被显著抑制，并且结肠直肠癌细胞被阻滞在G2-M期。在体内，通过pUbcH 10-RNAi下调UbcH 10基因的表达也抑制了裸鼠移植瘤的生长.我们的研究表明，RNA干扰介导的UbcH 10基因沉默对结直肠癌具有抗肿瘤活性，可能对结直肠癌的治疗具有潜在的治疗价值。
P>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.