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赵军, 路静, 杨洪艳, 黄幼田, 赵继敏, 赵明耀, 赵国强, 张曦, 董子明. DNA聚合酶β基因表达对NIH3T3细胞增殖影响[J]. 中国公共卫生, 2010, 26(1): 106-108. DOI: 10.11847/zgggws2010-26-01-57
引用本文: 赵军, 路静, 杨洪艳, 黄幼田, 赵继敏, 赵明耀, 赵国强, 张曦, 董子明. DNA聚合酶β基因表达对NIH3T3细胞增殖影响[J]. 中国公共卫生, 2010, 26(1): 106-108. DOI: 10.11847/zgggws2010-26-01-57
ZHAO Jun, LU Jing, YANG Hong-yan, . Effect of different type DNA polymerase β expression on proliferation of NIH3T3 cell line[J]. Chinese Journal of Public Health, 2010, 26(1): 106-108. DOI: 10.11847/zgggws2010-26-01-57
Citation: ZHAO Jun, LU Jing, YANG Hong-yan, . Effect of different type DNA polymerase β expression on proliferation of NIH3T3 cell line[J]. Chinese Journal of Public Health, 2010, 26(1): 106-108. DOI: 10.11847/zgggws2010-26-01-57

DNA聚合酶β基因表达对NIH3T3细胞增殖影响

Effect of different type DNA polymerase β expression on proliferation of NIH3T3 cell line

  • 摘要: 目的 观察不同类型DNA聚合酶β(DNA Polymerase β,DNA pol β)在小鼠成纤维细胞(NIH3T3细胞)的表达及细胞增殖特性的变化。方法 用脂质体转染的方法,将野生型(wild type,wt)和食管癌缺失突变型(delete mutional type,dmt)的polβ重组荧光载体pEGFP-C3-polβ转染NIH3T3细胞,建立稳定表达polβ的细胞系,通过荧光倒置显微镜观察其定位,四甲基偶氮噻唑蓝(MTT)比色法绘制生长曲线,流式细胞仪测定其细胞周期。结果 转染2种类型polβ的NIH3T3细胞株中,均有外源基因mRNA水平和蛋白水平的表达;荧光倒置显微镜结果显示,野生型的以细胞胞核为主,缺失突变型的polβ表达以细胞胞浆为主,从第3d开始,缺失突变型的细胞生长速度较野生型和对照组明显增快(P<0.05);转染缺失突变型NIH3T3细胞的S期细胞比例为68.29%,明显高于野生型的57.18%、对照组的58.12%和未转染NIH3T3细胞的57.35%(P<0.05)。结论 外源野生型polβ基因表达后对NIH3T3细胞的增殖速度无明显影响,而缺失突变型则使其增殖速度加快。

     

    Abstract: Objective To observe the proliferation characteristics of NIH3T3 cell line under expression of different type DNA polymerase B(DNA poBl).Methods The wild pol β and deletemutational pol β of esophageal cancer cellswere transfected into NIH3T3 cell line by lipofectamine method.The stable expression pol β NIH3T3 line was constructed.The location of pol β gene-encoded protein was observed with fluo rescent microscope.The growth curve was drawn with MTT method and the cell cycle was examined with flow cytomete.rResults It was proved that two pol β genes were transfected into the NIH3T3 cell line successfully.The location of wild type pol β prote in was mostly inside the nuclear,but the delete mutational pol β prote n was distributed in the whole cell.The growspeed of NIH3T3-wild type(wt)pol β was not obviously different compared with control cells and NIH3T3-EGFP(P>0.05).But the grow speed of NIH3T3-delete mutational type (dmt)tpol β was faster obviously(P<0.05).Furthermore,the cell account of Speriod of NIH3T3-dmtpol β was obviously increased(P<0.05),but that of NIH3T3-wt pol β showed no obvious change(P>0.05).Conclusion Expression of exogenous wild pol β is no obvious effect on the proliferation of NIH3T 3 cell and the expression of exogenous delete mutational po β can make NIH3T3 cells proliferate.There sult has great significance to further dicuss the pathogenesis of esophageal cancer and provides a strong basis for esophageal cancer biotherapy.

     

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