Department of Hepatology, Yantai Infectious Disease Hospital, Yantai, Shandong 264001, P.R. China.
Department of Research and Education, Yantai Infectious Disease Hospital, Yantai, Shandong 264001, P.R. China.
Mol Med Rep. 2019 Sep;20(3):2694-2702. doi: 10.3892/mmr.2019.10514. Epub 2019 Jul 23.
Complement factor H‑related 3 (CFHR3) belongs to the human factor H protein family and is associated with various human diseases, including nephropathy, age‑related macular degeneration and atypical hemolytic uremic syndrome. However, to the best of our knowledge, the role of CFHR3 in hepatocellular carcinoma (HCC) remains largely unknown. In the present study, reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) and western blot analysis were performed to determine mRNA and protein expression levels of CFHR3 in HCC and normal adjacent tissue. In addition, CFHR3 was overexpressed in Huh‑7 cells and cell counting kit‑8 assay was used to determine cell viability. Cell proliferation and apoptosis were assessed using flow cytometry, RT‑qPCR and western blotting. The results demonstrated that mRNA (2‑ΔΔCq) and protein expression levels of CFHR3 were significantly lower in tumor tissue compared with in adjacent tissue. Additionally, CFHR3 overexpression decreased cell viability, inhibited cell proliferation and significantly increased apoptosis. It was also identified that CFHR3 could downregulate the expression of Ki67. The results suggested that CFHR3 induced apoptosis by downregulating the expression of survivin and B cell lymphoma 2, upregulating the expression of Bcl‑2‑associated X and promoting caspase‑3 activity. Western blotting revealed that CFHR3 significantly inhibited the protein expression levels of phosphorylated (p)‑phosphoinositide 3‑kinase (PI3K), p‑protein kinase B (Akt) and p‑mammalian target of rapamycin (mTOR). Overexpression of CFHR3 suppressed proliferation and promoted apoptosis of HCC cells by inhibiting the PI3K/Akt/mTOR signaling pathway.
补体因子 H 相关蛋白 3(CFHR3)属于人补体因子 H 蛋白家族,与多种人类疾病相关,包括肾病、年龄相关性黄斑变性和非典型溶血性尿毒症综合征。然而,据我们所知,CFHR3 在肝细胞癌(HCC)中的作用仍知之甚少。在本研究中,通过逆转录-定量聚合酶链反应(RT-qPCR)和蛋白质印迹分析检测 CFHR3 在 HCC 和正常相邻组织中的 mRNA 和蛋白表达水平。此外,在 Huh-7 细胞中转染 CFHR3 过表达载体,并使用细胞计数试剂盒-8 测定细胞活力。通过流式细胞术、RT-qPCR 和蛋白质印迹分析评估细胞增殖和凋亡。结果表明,与相邻组织相比,肿瘤组织中 CFHR3 的 mRNA(2-ΔΔCq)和蛋白表达水平显著降低。此外,CFHR3 过表达降低细胞活力,抑制细胞增殖,显著增加细胞凋亡。还发现 CFHR3 可以下调 Ki67 的表达。结果表明,CFHR3 通过下调存活素和 B 细胞淋巴瘤 2 的表达、上调 Bcl-2 相关 X 的表达并促进半胱氨酸天冬氨酸蛋白酶-3 活性来诱导细胞凋亡。蛋白质印迹分析显示,CFHR3 显著抑制磷酸化(p)-磷酸肌醇 3-激酶(PI3K)、p-蛋白激酶 B(Akt)和 p-哺乳动物雷帕霉素靶蛋白(mTOR)的蛋白表达水平。CFHR3 过表达通过抑制 PI3K/Akt/mTOR 信号通路抑制 HCC 细胞的增殖并促进其凋亡。