Shen Mo, Zhou Lianlian, Zhou Ping, Zhou Wu, Lin Xiangyang
Department of Laboratory Medicine, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, P.R. China.
Department of Laboratory Medicine, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, P.R. China.
Mol Med Rep. 2017 Jul;16(1):937-942. doi: 10.3892/mmr.2017.6676. Epub 2017 Jun 1.
The role of inflammation in tumorigenesis and development is currently well established. Lymphotoxin β receptor (LTβR) activation induces canonical and noncanonical nuclear factor (NF)‑κB signaling pathways, which are linked to inflammation‑induced carcinogenesis. In the present study, 5,637 bladder cancer cells were cultured and the activation of LTβR was induced by functional ligand, lymphotoxin (LT) α1β2, and silencing with shRNA. Reverse transcription‑quantitative polymerase chain reaction was utilized to detect the mRNA expression levels of NF‑κB family members RelA and RelB, cytokines including LTα, LTβ, tumor necrosis factor (TNF)α, TNF superfamily member 14, interleukin (IL)‑6 and IL‑1β, and proliferation‑related genes including CyclinD1 and Survivin. The expression of phospho‑p65 was determined by western blotting. Activation of LTβR on bladder cancer 5,637 cells was demonstrated to upregulate the mRNA expression levels of the RELA proto‑oncogene, RelA, by 2.5‑fold compared with unstimulated cells, while no significant change was observed in the RELB proto‑oncogene NF‑κB member mRNA levels. Expression of pro‑inflammatory cytokines tumor necrosis factor (TNF)α and interleukin (IL)‑1β mRNA levels were significantly increased nearly 5‑fold and 1.5‑fold, respectively, following LTβR activation compared with unstimulated cells. The LTβR‑induced upregulation of RelA, TNFα and IL‑1β was decreased by ~33, 27, and 26% respectively when LTβR was silenced via short hairpin RNA. Activation of LTβR had no effect on 5,637 cell growth, despite CyclinD1 and Survivin mRNA levels increasing by ~2.7 and 1.3‑fold, respectively, compared with unstimulated cells. In conclusion, activation of LTβR induced the expression of RelA mRNA levels. LTβR activation might be an important mediator in promoting an inflammatory microenvironment in bladder cancer, via the upregulation of TNFα and IL‑1β mRNA levels. LTβR may be a potential therapeutic target for bladder cancer.
炎症在肿瘤发生和发展中的作用目前已得到充分证实。淋巴毒素β受体（LTβR）激活可诱导经典和非经典核因子（NF）-κB信号通路，这与炎症诱导的致癌作用有关。在本研究中，培养了5637个膀胱癌细胞，通过功能性配体淋巴毒素（LT）α1β2诱导LTβR激活，并使用短发夹RNA（shRNA）进行沉默。利用逆转录定量聚合酶链反应检测NF-κB家族成员RelA和RelB、细胞因子（包括LTα、LTβ、肿瘤坏死因子（TNF）α、TNF超家族成员14、白细胞介素（IL）-6和IL-1β）以及增殖相关基因（包括细胞周期蛋白D1和生存素）的mRNA表达水平。通过蛋白质印迹法测定磷酸化p65的表达。结果表明，与未刺激的细胞相比，膀胱癌细胞5637上的LTβR激活可使RELA原癌基因RelA的mRNA表达水平上调2.5倍，而RELB原癌基因NF-κB成员的mRNA水平未观察到显著变化。与未刺激的细胞相比，LTβR激活后促炎细胞因子肿瘤坏死因子（TNF）α和白细胞介素（IL）-1β的mRNA水平分别显著增加了近5倍和1.5倍。当通过短发夹RNA沉默LTβR时，LTβR诱导的RelA、TNFα和IL-1β上调分别降低了约33%、27%和26%。尽管与未刺激的细胞相比，细胞周期蛋白D1和生存素的mRNA水平分别增加了约2.7倍和1.3倍，但LTβR激活对5637细胞生长没有影响。总之，LTβR激活诱导了RelA mRNA水平的表达。LTβR激活可能是通过上调TNFα和IL-1β的mRNA水平来促进膀胱癌炎症微环境的重要介质。LTβR可能是膀胱癌的一个潜在治疗靶点。
