Liu Pingyi, Chen Lingling
Department of Haematology Laboratory, Linyi Central Hospital, Linyi 276400, Shandong, China.
Department of Haematology Laboratory, Linyi Central Hospital, Linyi 276400, Shandong, China.
Leuk Res. 2018 Jan;64:1-9. doi: 10.1016/j.leukres.2017.11.001. Epub 2017 Nov 11.
The present study explored the effects of Forkhead box Q1 (FOXQ1) on cell proliferation, cell cycle and apoptosis via the Sonic hedgehog (Shh) pathway in Natural killer/T-cell lymphoma (NKTCL). Quantitative real time-polymerase chain reaction (qRT-PCR) was performed to detect FOXQ1 expression in 117 NKTCL patients and 120 healthy controls. Additionally, FOXQ1 expression in NKTCL cell lines (HANK-1, NK-92, SNK-6, SNT-8 and YT) was determined by western blotting and qRT-PCR. SNK-6 cells were transfected with FOXQ1-shRNA or Shh pathway inhibitor Cyclopamine/recombinant protein Shh. Cell counting kit-8 (CCK-8) and 5-bromo-2-deoxy-uridine (BrdU) incorporation assays were conducted to detect cell proliferation, flow cytometry was used to determine the cell cycle and cell apoptosis, and western blotting was used to detect protein expression. FOXQ1 expression was higher in NKTCL patients than in healthy controls, which was related to Ann Arbor stage, bone marrow involvement and the 5year survival rate in NKTCL patients. Moreover, FOXQ1 expression, pathological type, Ann Arbor stage, B symptom and bone marrow involvement were independent risk factors in NKTCL. Shh pathway-related proteins were down-regulated after transfection of SNK-6 cells with FOXQ1-shRNA. Additionally, SNK-6 cell proliferation was greatly reduced, the cell cycle was blocked at the G0/G1 phase, and the expression of CyclinD1 and CyclinE was markedly decreased, while an increase in cell apoptosis with elevated Bcl-2-associated X protein (Bax) and Caspase-3 and reduced B-cell lymphoma/leukemia-2 (Bcl-2) were also observed. However, no significant alterations were observed between the FOXQ1-shRNA+Shh and Blank groups. The inhibition of FOXQ1 restricted NKTCL cell proliferation and growth but induced apoptosis via blocking the Shh signaling pathway.
本研究探讨了叉头框Q1（FOXQ1）通过音猬因子（Shh）信号通路对自然杀伤/T细胞淋巴瘤（NKTCL）细胞增殖、细胞周期及凋亡的影响。采用定量实时聚合酶链反应（qRT-PCR）检测117例NKTCL患者及120例健康对照者中FOXQ1的表达。此外，通过蛋白质免疫印迹法和qRT-PCR检测NKTCL细胞系（HANK-1、NK-92、SNK-6、SNT-8和YT）中FOXQ1的表达。用FOXQ1-shRNA或Shh信号通路抑制剂环杷明/重组蛋白Shh转染SNK-6细胞。采用细胞计数试剂盒-8（CCK-8）法和5-溴-2'-脱氧尿苷（BrdU）掺入法检测细胞增殖，流式细胞术检测细胞周期及细胞凋亡，蛋白质免疫印迹法检测蛋白表达。NKTCL患者中FOXQ1表达高于健康对照者，且与NKTCL患者的Ann Arbor分期、骨髓受累情况及5年生存率相关。此外，FOXQ1表达、病理类型、Ann Arbor分期、B症状及骨髓受累是NKTCL的独立危险因素。用FOXQ1-shRNA转染SNK-6细胞后，Shh信号通路相关蛋白表达下调。此外，SNK-6细胞增殖明显降低，细胞周期阻滞于G0/G1期，细胞周期蛋白D1（CyclinD1）和细胞周期蛋白E（CyclinE）表达明显下降，同时观察到细胞凋亡增加，Bcl-2相关X蛋白（Bax）和半胱天冬酶-3（Caspase-3）表达升高，B细胞淋巴瘤/白血病-2（Bcl-2）表达降低。然而，FOXQ1-shRNA+Shh组与空白组之间未观察到显著差异。抑制FOXQ1可通过阻断Shh信号通路限制NKTCL细胞增殖和生长，但诱导细胞凋亡。
