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Knockdown of GCF2/LRRFIP1 by RNAi Causes Cell Growth Inhibition and Increased Apoptosis in Human Hepatoma HepG2 Cells

机译:RNAI的GCF2 / LRRFIP1的敲低导致细胞生长抑制和增加人肝癌Hepg2细胞的细胞凋亡

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GC-binding factor 2 (GCF2) is a transcriptional regulator that represses transcriptional activity of the epidermal growth factor receptor (EGFR) by binding to a specific GC-rich sequence in the EGFR gene promoter. In addition to this function, GCF2 has also been identified as a tumor-associated antigen and regarded as a potentially valuable serum biomarker for early human hepatocellular carcinoma (HCC) diagnosis. GCF2 is high expressed in most HCC tissues and cell lines including HepG2. This study focused on the influence of GCF2 on cell proliferation and apoptosis in HepG2 cells. GCF2 expression at both mRNA and protein levels in HepG2 cells was detected with reverse transcription (RT) PCR and Western blotting, respectively. RNA interference (RNAi) technology was used to knock down GCF2 mRNA and protein expression. Afterwards, cell viability was analyzed with a Cell Counting Kit-8 (CCK-8), and cell apoptosis and caspase 3 activity by flow cytometry and with a Caspase 3 Activity Kit, respectively. Specific down-regulation of GCF2 expression caused cell growth inhibition, and increased apoptosis and caspase 3 activity in HepG2 cells. These primary results suggest that GCF2 may influence cell proliferation and apoptosis in HepG2 cells, and also provides a molecular basis for further investigation into the possible mechanism at proliferation and apoptosis in HCC.
机译:GC结合因子2(GCF2)是通过与EGFR基因启动子中的特异性GC的富序列结合来压制表皮生长因子受体(EGFR)的转录活性的转录调节剂。除此功能外,GCF2还被鉴定为肿瘤相关的抗原,并被视为用于早期人肝细胞癌(HCC)诊断的潜在有价值的血清生物标志物。 GCF2在大多数HCC组织和包括HepG2的细胞系中表达高。本研究重点是GCF2对HepG2细胞细胞增殖和细胞凋亡的影响。用逆转录(RT)PCR和Western印迹检测HepG2细胞中mRNA和蛋白质水平的GCF2表达。 RNA干扰(RNAi)技术用于击退GCF2 mRNA和蛋白质表达。然后,用细胞计数试剂盒-8(CCK-8)和细胞凋亡和Caspase 3分别通过流式细胞术分析细胞活力,并分别用Caspase 3活性试剂盒进行分析。 GCF2表达的具体下调引起了细胞生长抑制,并增加了HepG2细胞中的细胞凋亡和胱天蛋白酶3活性。这些主要结果表明,GCF2可能影响HepG2细胞中细胞增殖和凋亡,并提供分子基础,以进一步调查HCC中增殖和细胞凋亡的可能机制。

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