Gao Weilu, Hu Yong, Zhang Zhengqin, Du Gongwen, Yin Li, Yin Zongsheng
Department of Orthopaedics, The First Affiliated Hospital of Anhui Medical University, Hefei 230022, China,
Department of Anesthesiology, The First Affiliated Hospital of Anhui Medical University, Hefei 230022, China.
Onco Targets Ther. 2019 Feb 14;12:1225-1235. doi: 10.2147/OTT.S187209. eCollection 2019.
As a component of the EIF3 complex, EIF3C is essential for several steps in protein synthesis initiation. Recently, it has been addressed that EIF3C is overexpressed in several human cancers and plays a pivotal role in cell proliferation and tumorigenesis.
Immunohistochemistry, quantitative real-time PCR (qPCR), and Western blotting assays were employed to determine the expression of EIF3C in osteosarcoma (OsC) tissues obtained from 60 patients. The levels of EIF3C mRNA and protein were assessed by qPCR and Western blotting, respectively. The effect of EIF3C knockdown on OsC cell proliferation was detected by MTT and colony formation assays, respectively. Cell apoptosis induced by EIF3C silencing was analyzed by flow cytometric analysis. PathScan stress and apoptosis signaling antibody array kit was used to analyze the potential effects of EIF3C knockdown on OsC cells.
The levels of EIF3C were high in OsC tissues and cell lines. In addition, EIF3C knockdown by lentivirus-mediated shRNA targeting EIF3C significantly suppressed cell proliferation and colony formation and induced apoptosis in U-2OS cells. Moreover, EIF3C knockdown led to the upregulated expression of CASP3/7, Chk1/2, and SAPK/JNK, indicating that the downregulated expression of EIF3C might be associated with pro-apoptosis of U-2OS cells.
EIF3C may be a promising target for gene therapy of human OsC. However, the precise mechanisms behind the effect of EIF3C on OsC tumorigenesis require further analysis.
作为真核生物翻译起始因子3（EIF3）复合物的一个组成部分，EIF3C在蛋白质合成起始的多个步骤中至关重要。最近，有研究表明EIF3C在几种人类癌症中过表达，并在细胞增殖和肿瘤发生中起关键作用。
采用免疫组织化学、定量实时聚合酶链反应（qPCR）和蛋白质免疫印迹分析，以确定从60例患者获取的骨肉瘤（OsC）组织中EIF3C的表达情况。分别通过qPCR和蛋白质免疫印迹分析评估EIF3C信使核糖核酸（mRNA）和蛋白质水平。分别采用噻唑蓝（MTT）法和集落形成试验检测EIF3C基因敲低对OsC细胞增殖的影响。通过流式细胞术分析EIF3C沉默诱导的细胞凋亡。使用PathScan应激和凋亡信号抗体阵列试剂盒分析EIF3C基因敲低对OsC细胞的潜在影响。
EIF3C在OsC组织和细胞系中表达水平较高。此外，通过慢病毒介导的靶向EIF3C的短发夹RNA（shRNA）敲低EIF3C，可显著抑制U-2OS细胞的增殖和集落形成，并诱导细胞凋亡。此外，EIF3C基因敲低导致半胱天冬酶3/7（CASP3/7）、细胞周期检测点激酶1/2（Chk1/2）和应激活化蛋白激酶/应激活化蛋白激酶（SAPK/JNK）的表达上调，表明EIF3C表达下调可能与U-2OS细胞的促凋亡作用有关。
EIF3C可能是人类骨肉瘤基因治疗的一个有前景的靶点。然而，EIF3C对骨肉瘤肿瘤发生作用背后的确切机制需要进一步分析。
