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

• Published in: Clinical and experimental pharmacology & physiology 2010-05, Vol.37 (5-6), p.525-529
• Main Authors: Chen, Shi-Min, Jiang, Chun-Ying, Wu, Jian-Ying, Liu, Bin, Chen, Ying-Jian, Hu, Cheng-Jin, Liu, Xian-Xi
• Format: Article
• Language: English
• Subjects: Animals; Blotting, Western; Cell Division - genetics; colorectal cancer; Colorectal Neoplasms - enzymology; Colorectal Neoplasms - genetics; Colorectal Neoplasms - therapy; Down-Regulation; G2 Phase - genetics; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Neoplastic; Gene Silencing; gene therapy; Humans; Mice; Mice, Nude; nude mouse xenografts assay; Reverse Transcriptase Polymerase Chain Reaction; RNA Interference; Transfection; UbcH10; Ubiquitin-Conjugating Enzymes - genetics; Xenograft Model Antitumor Assays
• ISSN: 0305-1870; 1440-1681
• DOI: 10.1111/j.1440-1681.2010.05348.x

Summary 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.