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DNA聚合酶β基因表达对NIH3T3细胞增殖影响
  • ISSN号:1001-0580
  • 期刊名称:《中国公共卫生》
  • 时间:0
  • 分类:R73-362[医药卫生—肿瘤;医药卫生—临床医学]
  • 作者机构:[1]郑州大学基础医学院病理生理学教研室,郑州450052, [2]长治医学院附属和济医院内科, [3]郑州大学第三附属医院妇产科
  • 相关基金:国家自然科学基金(30471952)
中文摘要:

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。

英文摘要:

Objective To observe the proliferation characteristics of NIH3T3 cell line under expression of different type DNA polymerase β (DNA polβ).Methods The wild polβ and delete mutational polβ of esophageal cancer cells were transfected into NIH3T3 cell line by lipofectamine method.The stable expression polβ NIH3T3 line was constructed.The location of polβ gene-encoded protein was observed with fluorescent microscope.The growth curve was drawn with MTT method and the cell cycle was examined with flow cytometer.Results It was proved that two polβ genes were transfected into the NIH3T3 cell line successfully.The location of wild type polβ protein was mostly inside the nuclear,but the delete mutational polβ protein was distributed in the whole cell.The grow speed of NIH3T3-wild type(wt) polβ was not obviously different compared with control cells and NIH3T3-EGFP(P0.05).But the grow speed of NIH3T3-delete mutational type(dmt) tpolβ was faster obviously(P0.05).Furthermore,the cell account of S period of NIH3T3-dmt polβ was obviously increased(P0.05),but that of NIH3T3-wt polβ showed no obvious change(P0.05).Conclusion Expression of exo-genous wild polβ is no obvious effect on the proliferation of NIH3T3 cell and the expression of exogenous delete mutational polβ can make NIH3T3 cells proliferate.The result has great significance to further dicuss the pathogenesis of esophageal cancer and provides a strong basis for esophageal cancer biotherapy.

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期刊信息
  • 《中国公共卫生》
  • 北大核心期刊(2011版)
  • 主管单位:国家卫生和计划生育委员会
  • 主办单位:中华预防医学会
  • 主编:王宇
  • 地址:沈阳市和平区砂阳路242号
  • 邮编:110005
  • 邮箱:zgggws@vip.sina.com
  • 电话:024-23388443 23388479
  • 国际标准刊号:ISSN:1001-0580
  • 国内统一刊号:ISSN:21-1234/R
  • 邮发代号:8-204
  • 获奖情况:
  • 1999年获中华预防医学会系列期刊优秀期刊二等奖,2000年获中华预防医学会系列期刊优秀期刊三等奖,国家期刊提名奖
  • 国内外数据库收录:
  • 美国化学文摘(网络版),中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),中国北大核心期刊(2000版)
  • 被引量:66800