Wang Yingchao, Ma Lina, Wang Chunmei, Sheng Guangyao, Feng Lei, Yin Chuyun
Department of Pediatrics, the First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450000, P.R. China.
Int J Mol Med. 2015 Sep;36(3):627-32. doi: 10.3892/ijmm.2015.2267. Epub 2015 Jun 30.
The aberrant activation of autocrine motility factor receptor (AMFR) has been implicated in several types of human cancer. The present study aimed to elucidate the effect of AMFR on the regulation of proliferation in an acute monocytic leukemia cell line, THP‑1. THP‑1 cells were transfected with AMFR‑targeted small interfering (si)RNA and a plasmid encoding a truncated AMFR, AMFR‑C, (pcDNA3.1‑AMFR‑C). The mRNA and protein levels of AMFR and the downstream targets, rho‑associated, coiled‑coil containing protein kinase 2 (ROCK2), cyclin D1, and B‑cell lymphoma (Bcl)‑2, were measured using reverse transcription‑quantitatibe polymerase chain reaction and immunoblot analyses. The effects on cell cycle and apoptosis were investigated using flow cytometry. The present study successfully established the knockdown of AMFR and expression of AMFR‑C in the THP‑1 cells. Downregulation of AMFR induced cell cycle arrest at the G0/G1 phase, and increased apoptosis of the THP‑1 cells (all P<0.05). The AMFR siRNA increased the percentage of early apoptotic cells between 3.88±1.43 and 19.58±4.29% (P<0.05). The expression levels of ROCK2, cyclin D1 and Bcl‑2 were reduced by the downregulation of AMFR and enhanced by overexpression of AMFR‑C. In conclusion, AMFR appears to be crucial for the proliferation of the THP‑1 acute monocytic leukemia cell line. Therefore, AMFR may represent a potential target for the treatment of acute monocytic leukemia.
自分泌运动因子受体（AMFR）的异常激活与多种人类癌症有关。本研究旨在阐明AMFR对急性单核细胞白血病细胞系THP-1增殖调控的影响。用靶向AMFR的小干扰（si）RNA和编码截短型AMFR即AMFR-C的质粒（pcDNA3.1-AMFR-C）转染THP-1细胞。采用逆转录定量聚合酶链反应和免疫印迹分析检测AMFR及其下游靶点——含 Rho 相关卷曲螺旋结构域蛋白激酶2（ROCK2）、细胞周期蛋白D1和B细胞淋巴瘤（Bcl）-2的mRNA和蛋白水平。通过流式细胞术研究对细胞周期和凋亡的影响。本研究成功在THP-1细胞中实现了AMFR的敲低和AMFR-C的表达。AMFR的下调诱导细胞周期停滞在G0/G1期，并增加了THP-1细胞的凋亡（均P<0.05）。AMFR siRNA使早期凋亡细胞百分比从3.88±1.43%增加到19.58±4.29%（P<0.05）。AMFR的下调降低了ROCK2、细胞周期蛋白D1和Bcl-2的表达水平，而AMFR-C的过表达则增强了它们的表达。总之，AMFR似乎对THP-1急性单核细胞白血病细胞系的增殖至关重要。因此，AMFR可能是治疗急性单核细胞白血病的一个潜在靶点。
