METHODS: MCF7 and MDA-MB-231 cell lines were cultured in DMEM medium. After counting cells and measuring viability, Palbociclib was administered at varying doses using the IC50 obtained from MTT, with the treatment given at two time points of 24 and 72 hours. RNA was extracted from untreated and treated cells and RNAs were converted to cDNA in the end. Gene expression changes were investigated by real-time PCR. Data management and analysis were conducted using GraphPad Prism 5.01 software.
CONCLUSIONS: Among investigated genes, E2F3 gene was not significantly affected by Palbociclib in any of cell lines and time points. Besides, the expression of CCNE1 gene was significantly suppressed. It seems this drug was unable to reduce the expression of MDM2 gene significantly in triple negative (MDA-MB-231) cancer cells; however, a decrease was observed in luminal A (MCF-7) cells. CDKN2A and miR-141 genes expression increased significantly after treatment which can be aligned with palbociclib in proliferation inhibition.
方法:MCF7和MDA-MB-231细胞系在DMEM培养基中培养。计数细胞和测量活力后,使用从MTT获得的IC50以不同剂量施用Palbociclib,在24和72小时的两个时间点给予治疗。从未处理和处理的细胞中提取RNA,最后将RNA转化为cDNA。通过实时PCR研究基因表达变化。使用GraphPadPrism5.01软件进行数据管理和分析。
结论:在研究的基因中,E2F3基因在任何细胞系和时间点均未受到Palbociclib的显着影响。此外,CCNE1基因的表达被显著抑制。该药物似乎不能显著降低三阴性(MDA-MB-231)癌细胞中MDM2基因的表达;然而,在腔A(MCF-7)细胞中观察到减少。CDKN2A和miR-141基因表达在治疗后显著增加,这可以与palbociclib在增殖抑制方面一致。