Rna Interference-Mediated Silencing of Ubch10 Gene Inhibits Colorectal Cancer Cell Growth in Vitro and in Vivo

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.

Lentivirus-mediated RNA interference targeting UbcH10 reduces cell growth and invasion of human osteosarcoma cells via inhibition of Ki-67 and matrix metalloproteinases

“…This confirmed that the expression of UbcH10 is correlated with the proliferation activity of cancer cells. In accordance with the results of the present study, a previous study demonstrated that a knockdown of UbcH10 inhibited the cellular proliferation of other cancer cells, including those of lung, glioma and colorectal cancers (13)(14)(15). It is therefore hypothesized that OS cell growth inhibition is caused by UbcH10-mediated cell cycle regulation.…”

“…An overexpression of UbcH10 is significantly associated with the pathological grading of tumors, high cellular proliferation and poor prognoses of cancers affecting the adrenal gland, breast, colon, liver, lung and ovary (10,11). Furthermore, UbcH10 transgenic mice are prone to developing a range of spontaneous and carcinogen-induced tumors (12 (13,14). However, limited evidence exists regarding the biological function of UbcH10 in OS.…”

UbcH10 overexpression increases carcinogenesis and blocks ALLN susceptibility in colorectal cancer

“…Using these cells, we demonstrated that the inhibition of UbcH10 potently retarded the proliferation of tumor cells. Chen et al obtained similar results via RNA interference-mediated silencing of UbcH1022. Importantly, cell cycle changes and the accumulation of cyclin A and cyclin B1 were directly tested in endogenous conditions.…”

Increased Expression of Cellular Repressor of E1A-Stimulated Gene (CREG) in Gastric Cancer Patients: A Mechanism of Proliferation and Metastasis in Cancer

“…All the recombinant plasmids were confirmed by DNA sequencing. The MKN-45 cells were transfected with pGPU6/GFP/Neo-CREG siRNAs, and/or control siRNA using Lipofectamine TM 2000 reagent (Invitrogen), and were selected with G418 at the concentration of 500 lg/ml [9]. Subsequently, the knockdown efficiency was assessed via real-time RT-PCR and Western-blot analysis.…”