Department of Otolaryngology Head and Neck Surgery, Beijing Tongren Hospital, Capital Medical University, Beijing 100730, China.
Key Laboratory of Otolaryngology Head and Neck Surgery (Ministry of Education of China), Beijing Institute of Otolaryngology, Beijing 100005, China.
Chin Med J (Engl). 2020 Sep 5;133(17):2037-2043. doi: 10.1097/CM9.0000000000000933.
Long non-coding RNAs (lncRNAs) play key roles in human cancers. In our previous study, we demonstrated that lncRNA FKBP prolyl isomerase 9 pseudogene 1 (FKBP9P1) was highly expressed in head and neck squamous cell cancer (HNSCC) tissues. However, its functional significance remains poorly understood. In the present study, we identify the role and potential molecular biologic mechanisms of FKBP9P1 in HNSCC.
Quantitative real-time polymerase chain reaction was used to detect the expression of FKBP9P1 in HNSCC tissues, matched adjacent normal tissues, human HNSCC cells (FaDu, Cal-27, SCC4, and SCC9), and human immortalized keratinocytes cell HaCaT (normal control). Cal-27 and SCC9 cells were transfected with sh-FKBP9P1-1, sh-FKBP9P1-2, and normal control (sh-NC) lentivirus. Cell counting kit-8 assay, colony formation assay, wound healing assay, and trans-well assay were used to explore the biologic function of FKBP9P1 in HNSCC cells. Furthermore, western blotting was used to determine the mechanism of FKBP9P1 in HNSCC progression. Chi-squared test was performed to assess the clinical significance among FKBP9P1 high-expression and low-expression groups. Survival analyses were performed using the Kaplan-Meier method and assessed using the log-rank test. The comparison between two groups was analyzed by Student t test, and comparisons among multiple samples were performed by one-way analysis of variance and a Bonferroni post hoc test.
FKBP9P1 expression was significantly up-regulated in HNSCC tissues (tumor vs. normal, 1.914 vs. 0.957, t = 7.746, P < 0.001) and cell lines (P < 0.01 in all HNSCC cell lines). Besides, the median FKBP9P1 expression of HNSCC tissues (1.677) was considered as the threshold. High FKBP9P1 level was correlated with advanced T stage (P = 0.022), advanced N stage (P = 0.036), advanced clinical stage (P = 0.018), and poor prognosis of HNSCC patients (overall survival, P = 0.002 and disease-free survival, P < 0.001). Knockdown of FKBP9P1 led to marked repression in proliferation, migration, and invasion of HNSCC cells in vitro (P all < 0.01). Mechanistically, silencing FKBP9P1 was observed to restrain the PI3K/AKT signaling pathway.
Silencing lncRNA FKBP9P1 represses HNSCC progression and inhibits PI3K/AKT (phosphatidylinositol 3 kinase/AKT Serine/Threonine Kinase) signaling in vitro. Therefore, FKBP9P1 could be a potential new target for the diagnosis and treatment of HNSCC patients.
长非编码 RNA（lncRNA）在人类癌症中发挥关键作用。在我们之前的研究中，我们证明 lncRNA FKBP 脯氨酰异构酶 9 假基因 1（FKBP9P1）在头颈部鳞状细胞癌（HNSCC）组织中高度表达。然而，其功能意义仍知之甚少。在本研究中，我们确定了 FKBP9P1 在 HNSCC 中的作用和潜在的分子生物学机制。
使用实时定量聚合酶链反应检测 HNSCC 组织、配对的相邻正常组织、人 HNSCC 细胞（FaDu、Cal-27、SCC4 和 SCC9）和人永生化角质形成细胞 HaCaT（正常对照）中 FKBP9P1 的表达。Cal-27 和 SCC9 细胞用 sh-FKBP9P1-1、sh-FKBP9P1-2 和正常对照（sh-NC）慢病毒转染。细胞计数试剂盒-8 检测、集落形成检测、划痕愈合检测和 Transwell 检测用于研究 FKBP9P1 在 HNSCC 细胞中的生物学功能。此外，Western blot 用于确定 FKBP9P1 在 HNSCC 进展中的机制。卡方检验用于评估 FKBP9P1 高表达和低表达组之间的临床意义。使用 Kaplan-Meier 方法进行生存分析，并使用对数秩检验进行评估。使用 Student t 检验比较两组之间的差异，使用单因素方差分析和 Bonferroni 事后检验比较多个样本之间的差异。
FKBP9P1 在 HNSCC 组织（肿瘤与正常组织，1.914 与 0.957，t=7.746，P<0.001）和细胞系中明显上调（所有 HNSCC 细胞系中 P<0.01）。此外，HNSCC 组织中 FKBP9P1 的中位数表达（1.677）被认为是阈值。高 FKBP9P1 水平与晚期 T 期（P=0.022）、晚期 N 期（P=0.036）、晚期临床分期（P=0.018）和 HNSCC 患者预后不良（总生存，P=0.002 和无病生存，P<0.001）相关。FKBP9P1 的敲低导致体外 HNSCC 细胞增殖、迁移和侵袭明显受到抑制（P 均<0.01）。机制上，观察到沉默 FKBP9P1 可抑制 PI3K/AKT（磷脂酰肌醇 3 激酶/AKT 丝氨酸/苏氨酸激酶）信号通路。
沉默 lncRNA FKBP9P1 抑制 HNSCC 进展，并抑制体外 PI3K/AKT 信号通路。因此，FKBP9P1 可能是 HNSCC 患者诊断和治疗的潜在新靶点。
