Lau Wai Hoe, Zhu Xiphias Ge, Ho Shamaine Wei Ting, Chang Shu Chun, Ding Jeak Ling
Department of Biological Sciences, Faculty of Science, National University of Singapore, Singapore 117543, Singapore.
Current addresses: Taipei Medical University, College for Medical Science and Technology, Taipei 110, Taiwan.
Oncotarget. 2017 May 16;8(20):32884-32904. doi: 10.18632/oncotarget.15862.
Activation of TLR3 stimulates cancer cell apoptosis and triggers secretion of inflammatory cytokines. PolyI:C, a TLR3 agonist, activates immune cells and regresses metastatic lung cancer in vivo. Although polyI:C reportedly kills lung carcinomas, the mechanism remains elusive. Here, we demonstrated that polyI:C suppressed the proliferation and survival of metastatic (NCI-H358 and NCI-H292) and non-metastatic (A549) lung cancer cells. Notably, A549, NCI-H292 and NCI-H358 which are inducible by polyI:C, expressed low-to-medium level of TLR3 protein, and were susceptible to polyI:C treatment. By contrast, NCI-H1299, which endogenously expresses high level of TLR3 protein, was insensitive to polyI:C. We showed that polyI:C stimulated pro-inflammatory cytokines associated with survival and metastasis in a cell type-specific manner. While A549 and NCI-H292 released high levels of IL6, IL8 and GRO, the NCI-H358 cells endogenously secretes abundant levels of these cytokines, and was not further induced by polyI:C. Thus, NCI-H358 was resistant to the inhibition of cytokine-dependent metastasis. NCI-H1299, which was unresponsive to polyI:C, did not produce any of the pro-inflammatory cytokines. Treatment of A549 with a combination of polyI:C and anti-IL6 antibody significantly decreased IL6 production, and enhanced polyI:C-mediated killing and suppression of oncogenicity and metastasis. While polyI:C stimulated the phosphorylation of STAT3 and JAK2, blockade of these proteins enhanced polyI:C-mediated suppression of survival and metastasis. Taken together, polyI:C alone provoked apoptosis of lung cancer cells that express low-to-medium levels of functional TLR3 protein. The combinatorial treatment with polyI:C and anti-IL6 enhanced polyI:C-mediated anticancer activities through IL6/JAK2/STAT3 signalling, and apoptosis via TLR3-mediated caspase 3/8 pathway.
Toll样受体3(TLR3)的激活可刺激癌细胞凋亡并引发炎性细胞因子的分泌。聚肌胞苷酸(PolyI:C)作为一种TLR3激动剂,可激活免疫细胞并使体内转移性肺癌消退。尽管据报道PolyI:C可杀死肺癌细胞,但其机制仍不清楚。在此,我们证明了PolyI:C可抑制转移性(NCI-H358和NCI-H292)和非转移性(A549)肺癌细胞的增殖与存活。值得注意的是,可被PolyI:C诱导的A549、NCI-H292和NCI-H358表达低至中等水平的TLR3蛋白,并且对PolyI:C处理敏感。相比之下,内源性表达高水平TLR3蛋白的NCI-H1299对PolyI:C不敏感。我们发现,PolyI:C以细胞类型特异性方式刺激与存活和转移相关的促炎性细胞因子。虽然A549和NCI-H292释放高水平的白细胞介素6(IL6)、白细胞介素8(IL8)和生长调节致癌基因蛋白(GRO),但NCI-H358细胞内源性分泌大量这些细胞因子,并且不会被PolyI:C进一步诱导。因此,NCI-H358对细胞因子依赖性转移的抑制具有抗性。对PolyI:C无反应的NCI-H1299不产生任何促炎性细胞因子。用PolyI:C和抗IL6抗体联合处理A549可显著降低IL6的产生,并增强PolyI:C介导的杀伤作用以及对致癌性和转移的抑制作用。虽然PolyI:C刺激信号转导和转录激活因子3(STAT3)和Janus激酶2(JAK2)的磷酸化,但对这些蛋白的阻断增强了PolyI:C介导的对存活和转移的抑制作用。综上所述,单独使用PolyI:C可诱导表达低至中等水平功能性TLR3蛋白的肺癌细胞凋亡。PolyI:C与抗IL6的联合治疗通过IL6/JAK2/STAT3信号传导增强了PolyI:C介导的抗癌活性,并通过TLR3介导的半胱天冬酶3/8途径诱导凋亡。