Inhibition of sonic hedgehog signaling blocks cell migration and growth but induces apoptosis via suppression of FOXQ1 in natural killer/T-cell lymphoma
Inhibition of sonic hedgehog signaling blocks cell migration and growth but induces apoptosis via suppression of FOXQ1 in natural killer/T-cell lymphoma
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DOI：
10.1016/j.leukres.2017.11.001
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发表时间：
2018-01-01
影响因子：
2.7
通讯作者：
Chen, Lingling
中科院分区：
文献类型：
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作者：
Liu, Pingyi;Chen, Lingling
本研究探讨Forkhead box Q1（FOXQ 1）对自然杀伤/T细胞淋巴瘤（NKTCL）细胞增殖、细胞周期和凋亡的影响。采用定量真实的时间聚合酶链反应（qRT-PCR）检测117例NKTCL患者和120例健康对照者FOXQ 1的表达。此外，通过蛋白质印迹和qRT-PCR测定NKTCL细胞系（HANK-1、NK-92、SNK-6、SNT-8和YT）中的FOXQ 1表达。用FOXQ 1-shRNA或Shh通路抑制剂环巴胺/重组蛋白Shh转染SNK-6细胞。采用CCK-8法和5-溴脱氧尿嘧啶核苷（BrdU）掺入法检测细胞增殖情况，流式细胞术检测细胞周期和细胞凋亡，Western blotting检测蛋白表达。FOXQ 1在NKTCL患者中的表达明显高于正常对照组，其表达与NKTCL患者的安阿伯分期、骨髓受累及5年生存率有关。FOXQ 1表达、病理类型、安阿伯分期、B症状和骨髓受累是NKTCL的独立危险因素。FOXQ 1-shRNA转染SNK-6细胞后，Shh通路相关蛋白表达下调。另外，SNK-6细胞的增殖能力明显降低，细胞周期阻滞于G 0/G1期，CyclinD 1和CyclinE的表达明显减少，细胞凋亡增加，Bcl-2相关X蛋白（Bax）和Caspase-3表达增加，B细胞淋巴瘤/白血病-2（Bcl-2）表达减少。然而，在FOXQ 1-shRNA + Shh组和空白组之间没有观察到显著变化。抑制FOXQ 1可抑制NKTCL细胞的增殖和生长，但可通过阻断Shh信号通路诱导细胞凋亡。
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 5 year 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.