敲低PES1基因抑制舌癌细胞的生长
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摘要
目的构建稳定表达PES1 shRNA的舌癌细胞系,研究敲低PES1基因对舌癌细胞生长的影响。方法在293T细胞中包装并获得敲低PES1基因的病毒,然后将病毒感染Tca8113、SCC6和SCC15 3种舌癌细胞并筛选稳定克隆,Western印迹检测PES1蛋白的表达水平;利用生长曲线检测敲低PES1对舌癌细胞生长的影响,流式细胞术检测敲低PES1对舌癌细胞周期的影响。结果成功构建稳定表达PES1 shRNA的舌癌细胞系,敲低PES1基因能够抑制舌癌细胞的生长,使细胞周期阻滞在G0/G1期,敲低PES1抑制cyclin D1的表达。结论敲低PES1抑制舌癌细胞的生长,诱导细胞周期阻滞在G0/G1期,抑制cyclin D1的表达,因此PES1可能成为舌癌基因治疗的靶标。
Objective To construct PES1 shRNA stable expression cell lines in tongue squamous cell carcinoma(TSCC) cells and to study the effect of knockdown of PES1 on the growth of TSCC cells.Methods Recombinant lentivirus carrying PES1 shRNA was packaged and obtained in 293 T cells.TSCC cells(Tca8113,SCC6 and SCC15) were infected with the lentivirus and selected for stable cells.PES1 expression was identified by Western blot.The effect of inhibition of PES1 on the growth and cell cycle of TSCC cells was detected by growth curve and flow cytometry.Results TSCC cells stably expressing PES1 shRNA were constructed.Knockdown of PES1 inhibited cell proliferation and induced cell cycle arrest at G0/ G1 phase.Knockdown of PES1 inhibited expression of cyclin D1 in TSCC cells.Conclusion Inhibition of PES1 results in reduced cell proliferation,cell cycle arrest at G0/ G1 phase and reduction of cyclin D1 expression in TSCC cells.PES1 may be a target for TSCC gene therapy.
Objective To construct PES1 shRNA stable expression cell lines in tongue squamous cell carcinoma(TSCC) cells and to study the effect of knockdown of PES1 on the growth of TSCC cells.Methods Recombinant lentivirus carrying PES1 shRNA was packaged and obtained in 293 T cells.TSCC cells(Tca8113,SCC6 and SCC15) were infected with the lentivirus and selected for stable cells.PES1 expression was identified by Western blot.The effect of inhibition of PES1 on the growth and cell cycle of TSCC cells was detected by growth curve and flow cytometry.Results TSCC cells stably expressing PES1 shRNA were constructed.Knockdown of PES1 inhibited cell proliferation and induced cell cycle arrest at G0/ G1 phase.Knockdown of PES1 inhibited expression of cyclin D1 in TSCC cells.Conclusion Inhibition of PES1 results in reduced cell proliferation,cell cycle arrest at G0/ G1 phase and reduction of cyclin D1 expression in TSCC cells.PES1 may be a target for TSCC gene therapy.
引文
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[9]李杰萍,张浩,杨树兴,等.利用不同方法检测RNAi抑制人pescadillo基因的表达[J].细胞与分子免疫学杂志,2007,23(11):1064-1065.
[10]Li J,Yu L,Zhang H,et al.Down-regulation of pescadillo inhibits proliferation and tumorigenicity of breast cancer cells[J].Cancer Sci,2009,100(12):2255-2260.
[11]邱蔚六.中国口腔颌面肿瘤学进展50年[J].中华口腔医学杂志,2002,37(3):161-164.
[2]Holzel M,Grimm T,Rohrmoser M,et al.The BRCT domain of mammalian PES1 is crucial for nucleolar localization and rRNA processing[J].Nucleic Acids Res,2007,35(3):789-800.
[3]Zhang H,Fang Y,Huang C,et al.Human pescadillo induces large-scale chromatin unfolding[J].Sci Chin C Life Sci,2005,48(3):270-276.
[4]Prisco M,Maiorana A,Guerzoni C,et al.Role of pescadillo and upstream binding factor in the proliferation and differentiation of murine myeloid cells[J].Mol Cell Biol,2004,24(12):5421-5433.
[5]Lerch-Gaggl A,Haque J,Li JX,et al.Pescadillo is essential for nucleolar assembly,ribosome biogenesis,and mammalian cell proliferation[J].J Biol Chem,2002,277(47):45347-45355.
[6]Siegel R,Naishadham D,Jemal A.Cancer statistic[J].CA Cancer J Clin,2012,62(24):10-29.
[7]Weber A,Hengge UR,Stricker I,et al.Protein microarrays for the detection of biomarkers in head and neck squamous cell carcinomas[J].Hum Pathol,2007,38(13):C228-C238.
[8]高卫栋,王红兵,吴泽志,等.肿瘤细胞迁移特性及细胞迁移能力表征[J].细胞生物学杂志,2008,30(4):451-456.
[9]李杰萍,张浩,杨树兴,等.利用不同方法检测RNAi抑制人pescadillo基因的表达[J].细胞与分子免疫学杂志,2007,23(11):1064-1065.
[10]Li J,Yu L,Zhang H,et al.Down-regulation of pescadillo inhibits proliferation and tumorigenicity of breast cancer cells[J].Cancer Sci,2009,100(12):2255-2260.
[11]邱蔚六.中国口腔颌面肿瘤学进展50年[J].中华口腔医学杂志,2002,37(3):161-164.