泛素偶联酶2C促进肺癌细胞增殖和迁移与抑制细胞衰老

UBE2C Promotes Proliferation and Migration of Lung Cancer Cells and Inhibits Cell Senescence

泛素偶联酶2C与多种肿瘤细胞的增殖密切相关,但其与肺癌发生和发展的关系尚不明确。本研究以肺癌A549细胞为材料,通过RT-PCR、Western印迹、免疫荧光、SA-β-Gal细胞衰老染色、细胞划痕和Trans-well实验,阐明UBE2C与肺癌细胞的增殖、衰老和迁移能力的关系。结果显示,UBE2C在肺癌细胞中的表达明显高于正常细胞。利用基因修饰技术瞬时过表达或靶向沉默UBE2C后,在肺癌A549细胞中,UBE2C的mRNA和蛋白质水平显著增加3.5倍或减少0.5倍,显著促进或抑制细胞增殖,进而减少或增加细胞的凋亡率。过表达UBE2C后,显著抑制细胞衰老;但沉默UBE2C后,则增加细胞衰老。此外,过表达UBE2C后,下调转移相关基因E-钙黏着蛋白的mRNA和蛋白质表达水平,且上调波形蛋白基因的表达水平,进而促进肺癌细胞的迁移。但靶向敲除UBE2C后,上调E-钙黏着蛋白,同时下调波形蛋白表达水平,进而抑制肺癌细胞的迁移。本研究的开展将明确UBE2C在肺癌中的作用及其机制,为以UBE2C为靶点,提高病人生存期提供了理论基础。

Ubiquitin-conjugating enzyme E2C（ UBE2C） is closely related to the proliferation of various tumor cells,but its relationship with the occurrence and development of lung cancer is not clear. The roles of UBE2C in cellular proliferation,cell senescence and migration of A549 lung cancer cells were elucidated by RT-PCR,Western blotting,immunofluorescence,SA-β-Gal cell senescence staining,cell scratch and Trans-well assays. Transient over-expression or targeted silencing of UBE2C by gene modification technique was used. The results showed that the expression of UBE2C in lung cancer cells was significantly higher than that in normal cells. The levels of UBE2C mRNA and protein were significantly increased by 3. 5-fold or decreased by 0. 5-fold in A549 cells after transient over-expression or silence of UBE2C,leading to significant promotion or inhibition of cell proliferation,thereby reducing or increasing the rate of apoptosis. In addition,over-expression of UBE2C significantly inhibited cell senescence,whereas silence of UBE2C promoted cell senescence. Moreover, UBE2C significantlyaffected the mRNA and protein expression of metastasis related genes E-cadherin and vimentin,and further modulated cell migration. This study provided information for the roles of UBE2C in the development and progression of lung cancer.