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目的探讨泛素结合酶-10(Ubiquitin-conjugating enzymes,UbcH10)表达对结直肠癌细胞增殖的影响。方法采用脂质体LipofectamineTM2000转染人UbcH10的真核表达载体pcDNA3.1-UbcH10及其对照载体pcDNA3.1至结直肠癌细胞LoVo,蛋白印迹(Western blot)技术分析UbcH10的表达;细胞增殖试验(CCK-8试剂)检测UbcH10表达对LoVo细胞增殖的影响;流式细胞术检测UbcH10表达对LoVo细胞周期的影响。结果 Westernblot分析证实,与对照组细胞相比,转染重组质粒的LoVo细胞中UbcH10表达明显升高;细胞生长增殖曲线显示,UbcH10过度表达的细胞生长明显快于对照组细胞。流式细胞术分析细胞周期结果显示:与对照组相比,UbcH10过度表达组细胞在G2-M期分布明显减少(P<0.05)。结论 UbcH10能促进结直肠癌细胞增殖,调控细胞周期进展可能是其潜在的作用机制。
Objective To investigate the effect of ubiquitin-conjugating enzymes (UbcH10) on the proliferation of colorectal cancer cells. Methods LipofectamineTM2000 was used to transfect human UbcH10 eukaryotic expression vector pcDNA3.1-UbcH10 and its control vector pcDNA3.1 into colorectal cancer cells LoVo, Western blot analysis of UbcH10 expression; cell proliferation assay ( CCK-8 reagent) to detect the effect of UbcH10 expression on LoVo cell proliferation; Flow cytometry was used to detect the effect of UbcH10 expression on LoVo cell cycle. Results Western blot analysis confirmed that the expression of UbcH10 in LoVo cells transfected with recombinant plasmid was significantly higher than that in control cells. The proliferation and proliferation curve of UbcH10 overexpressing cells showed that UbcH10 overexpression was faster than that of control cells. Cell cycle analysis by flow cytometry showed that the distribution of UbcH10 overexpression cells in G2-M phase was significantly decreased (P <0.05) compared with the control group. Conclusion UbcH10 can promote the proliferation of colorectal cancer cells and regulate the cell cycle progression, which may be a potential mechanism of action.