肿瘤坏死因子-α 通过激活 VEGFC 介导的 AKT 和 ERK 通路促进宫颈癌的发生、淋巴管生成和淋巴转移。

BACKGROUND: Lymphatic metastasis is a common phenomenon of cervical cancer. Tumor necrosis factor- (TNF-) was found to be closely associated with lymphatic cancer metastasis. However, the mechanism through which TNF- regulates lymphatic metastasis in cervical cancer remains unclear. METHODS: In this study, cervical cancer cells were cultured in Dulbecco's modified Eagle's medium (DMEM) with or without TNF- for 48 h, and then the corresponding conditional medium (CM-TNF- or CM) was collected. The level of vascular endothelial growth factor (VEGFC) in the corresponding CM was then detected using an enzyme-linked immunosorbent assay (ELISA). Next, human lymphatic endothelial cells (HLECs) were cultured in CM-TNF- or CM for 48 h. Cell viability was measured using the cell counting kit-8 (CCK-8) assay, and angiogenesis was detected using a tube formation assay. Subsequently, the expressions of AKT, p-AKT, ERK, and p-ERK in HLECs were detected using western blotting. In addition, to further investigate the effect of TNF- on the progression of cervical cancer, a C33A subcutaneous xenograft model was established . RESULTS: We found that TNF- significantly stimulated cervical cancer cells to secrete VEGFC. Additionally, the CM collected from the TNF--treated cervical cancer cells notably promoted the proliferation, migration, and angiogenesis of HLECs; however, these changes were reversed by MAZ51, a VEGFR3 inhibitor. Moreover, TNF- obviously elevated D2-40 and VEGFC protein expressions in tumor tissues, promoting lymphangiogenesis and lymphatic metastasis . Meanwhile, TNF- markedly upregulated p-AKT and p-ERK expressions in tumor tissues, whereas these changes were reversed by MAZ51. CONCLUSION: Collectively, TNF- could promote tumorigenesis, lymphangiogenesis, and lymphatic metastasis and in cervical cancer via activating VEGFC-mediated AKT and ERK pathways. These results may provide new directions for the treatment of cervical cancer.

背景：淋巴转移是宫颈癌的常见现象。肿瘤坏死因子-（TNF-）被发现与癌症的淋巴转移密切相关。然而，TNF-调节宫颈癌淋巴转移的机制尚不清楚。

方法：在这项研究中，将宫颈癌细胞在含或不含 TNF-的 Dulbecco 改良 Eagle 培养基（DMEM）中培养 48 小时，然后收集相应的条件培养基（CM-TNF-或 CM）。使用酶联免疫吸附测定（ELISA）检测相应 CM 中的血管内皮生长因子（VEGFC）水平。接下来，将人淋巴管内皮细胞（HLECs）在 CM-TNF-或 CM 中培养 48 小时。使用细胞计数试剂盒-8（CCK-8）测定细胞活力，并通过管形成测定检测血管生成。随后，使用 Western blot 检测 HLECs 中 AKT、p-AKT、ERK 和 p-ERK 的表达。此外，为了进一步研究 TNF-对宫颈癌进展的影响，建立了 C33A 皮下移植瘤模型。

结果：我们发现 TNF-可显著刺激宫颈癌细胞分泌 VEGFC。此外，来自 TNF--处理的宫颈癌细胞的 CM 明显促进了 HLECs 的增殖、迁移和血管生成；然而，VEGFR3 抑制剂 MAZ51 逆转了这些变化。此外，TNF-明显提高了肿瘤组织中 D2-40 和 VEGFC 蛋白的表达，促进了淋巴管生成和淋巴转移。同时，TNF-明显上调了肿瘤组织中 p-AKT 和 p-ERK 的表达，而 MAZ51 逆转了这些变化。

结论：综上所述，TNF-可通过激活 VEGFC 介导的 AKT 和 ERK 通路促进宫颈癌的发生、淋巴管生成和淋巴转移。这些结果可能为宫颈癌的治疗提供新的方向。