摘要
[目的]探讨人皮肤表皮细胞(HaCaT)聚-ADP核糖聚合酶-1基因(PARP-1)过表达后的生物学性状变化。[方法]设试验组、空载体对照组、正常HaCaT对照组,运用真核表达载体使PARP-1基因在HaCaT中过表达,在光学显微镜下观察转染重组子的试验组细胞、转染空质粒的对照组细胞及正常对照组细胞的形态学特征。绘制细胞生长曲线,并运用流式细胞技术分析细胞周期。以苯并芘(BaP)恶性转化的细胞作为阳性对照,采用软琼脂克隆形成试验对细胞的恶性程度进行鉴定。[结果]光镜下试验组细胞与空载体组细胞及正常对照组细胞相比,除体积稍有缩小外,其他形态学特征没有明显变化。3组细胞的生长曲线均呈相似的S型,但转染重组子细胞的生长速度加快。试验组、空载体组及正常细胞组的细胞周期分布为G1期(44.3%、78.1%、76.4%)、G2期(3.6%、13.3%、12.2%)和S期(52.1%、8.6%、11.4%)。正常对照细胞和空载体组分布相近,试验组S期明显增多;试验组细胞未见在双层软琼脂糖中生长出细胞集落,而阳性对照组细胞集落明显。[结论]PARP-1基因过表达后可以改变HaCaT细胞的生长速度,但并不影响细胞的恶性程度。
[Objective] To explore the biological characteristics changes of PARP-1 overexpressed HaCaT cells.[Methods] Experimental group,empty vector group,normal HaCaT control group and positive control group were included.Morphological characteristics of each group were analyzed under light microscope.Growth curve of each group was detected by MTT assay.Flow cytomertry（FCM） assay was used to study the cell cycle distribution.Malignancy degree of each group was analyzed by soft agars experiment.[Result] In the experimental group,the numbers of cell granulations were increased transparently and cell volume decrease was found,the rest characteristics were common.Growth curve of each group presented similar shape of ＂S＂.The cell cycle distribution of G1（44.3%,78.1%,76.4%）,G2（3.6%,13.3%,12.2%） and S（52.1%,8.6%,11.4%） in the above mentioned groups appeared similar.In soft agarose experiment,cell colony was not found in the experimental group,but found in the positive control group.[Conclusion] Overexpressed PARP-1 gene can change the growth speed of HaCaT cells,but do not affect the grade malignancy of cells.
出处
《安徽农业科学》
CAS
2012年第1期205-207,共3页
Journal of Anhui Agricultural Sciences
基金
国家自然科学基金项目(30972505)
深圳市重点项目(2009-01017)
