摘要
目的:探讨CHFR与聚(ADP-核糖)聚合酶1(PARP-1)基因对B细胞淋巴瘤Raji细胞增殖和凋亡的影响。方法:用5-Aza-d C处理Raji细胞,后通过qRT-PCR检测CHFR的mRNA表达水平,western blot评估CHFR、PARP-1的蛋白表达。经CHFR慢病毒转染Raji细胞后,用qRT-PCR和western blot评估RAJI细胞中的CHFR、PARP-1变化。通过CCK-8法测定细胞的增殖情况,流式细胞术检测细胞周期的变化。结果:与对照组相比,经5-Aza-d C处理Raji组的CHFR mRNA表达显著上调(P<0.01),PARP-1mRNA水平和蛋白表达水平明显降低(P<0.05)。与对照组相比,Sh RNA组CHFR的mRNA表达显着下调(均P<0.01),PARP-1mRNA和蛋白表达水平升高(P<0.05)。CCK结果显示CHFR Sh RNA组细胞活力明显低于对照组(P<0.05)。流式细胞术结果显示CHFR沉默后细胞凋亡率降低(P<0.05)。结论:CHFR可能通过PARP-1调控B淋巴细胞的增殖和凋亡。
Objective： To investigate the effect of checkpoint with forkhead-associated （FHA） and RING finger domains （CHFR） with poly （ADP-ribose） polymerase 1 （PARP-1） on the proliferation and apoptosis of B-cell non-Hodgkin＇s lymphoma Raji cells. Methods： Quantitative real-time PCR （qRT-PCR） and western blot were used to detect the CHFR, PARP-1 mRNA and protein expressions in Raji cells treated by 5-Aza-dC and transfected by CHFR lentivirus. Cell proliferation was analyzed by a standard Cell Counting Kit-8 （CCK-8） assay, and the cell cycle changes were detected by flow cytometry. Results： The mRNA expression of CHFR in the 5-Aza-dC Raji group was significantly up-regulated compared with that in the control group （P 〈 0.01）. PARP-1 levels were decreased in mRNA levels and the protein expression levels compared with control group （all P〈0.05）. The mRNA expression of CHFR in the ShRNA group was significantly down-regulated compared with that in the control and normal group （both P 〈 0.01）. The mRNA and protein expression levels of PARP-1 were increased in ShRNA group compared with the control group and negative control group（P 〈 0.05）. The CCK results suggested that the decrease in CHFR expression promoted the cell proliferation （P〈0.05）. And Flow cytometry results demonstrated that the cell apoptosis rate was decreased after CHFR silencing （P〈0.05）. Conclusion： These data suggested that CHFR might regulate the proliferation and apoptosis of B-cell non-Hodgkin＇s lymphoma Raji cells through PARP-1.
出处
《现代生物医学进展》
CAS
2017年第8期1430-1436,共7页
Progress in Modern Biomedicine
基金
Qingdao Municipal Natural Science Foundation(2012-1-3-2-5-nsh)
Research Award Fund for Outstanding Middle-aged and Young Scientist of Shandong Province(BS2012YY004)
the Shandong Provincial Natural Science Foundation,China(Y2008C170,Q2008C04)~~
