Introduction

The prognosis for cervical cancer (CC) patients with lymph node metastasis (LNM) is extremely poor. Lipid droplets (LDs) have a pivotal role in promoting tumor metastasis. The crosstalk mechanism between LDs and LNM modulated in CC remains largely unknown.

Objectives

This study aimed to construct a miRNA-dependent progonostic model for CC patients and investigate whether miR-532-5p has a biological impact on LNM by regualting LDs accumulation.

Methods

LASSO-Cox regression was applied to establish a prognostic prediction model. miR-532-5p had the lowest P-value in RNA expression (P < 0.001) and prognostic prediction (P < 0.0001) and was selected for further study. The functional role of the prognostic miR-532-5p-correlated competing endogenous RNA (ceRNA) network was investigated to clarify the crosstalk between LDs and LNM. The underlying mechanism was determined using site-directed mutagenesis, dual luciferase reporter assays, RNA immunoprecipitation assays, and rescue experiments. A xenograft LNM model was established to evaluate the effect of miR-532-5p and orlistat combination therapy on tumor growth and LNM.

Results

A novel 5-miRNAs prognostic signature was constructed to better predict the prognosis of CC patient. Further study demonstrated that miR-532-5p inhibited epithelial-mesenchymal transition and lymphangiogenesis by regulating LDs accumulation. Interestingly, we also found that LDs accumulation promoted cell metastasis in vitro. Mechanistically, we demonstrated a miR-532-5p-correlated ceRNA network in which LINC01410 was bound directly to miR-532-5p and effectively functioned as miR-532-5p sponge to disinhibit its target gene-fatty acid synthase (FASN). Combined therapy with miR-532-5p and FASN inhibitor-orlistat further inhibited tumor growth and LNM in vivo.

Conclusion

Our findings highlight a LD accumulation-dependent mechanism of miR-532-5p-modulated LNM and support treatment with miR-532-5p/orlistat as novel strategy for treating patients with LNM in CC.

中文翻译：

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预后miR-532-5p相关的ceRNA介导的脂滴积累驱动宫颈癌的淋巴结转移

介绍

伴有淋巴结转移（LNM）的宫颈癌（CC）患者预后极差。脂滴（LDs）在促进肿瘤转移中具有关键作用。在 CC 中调制的 LD 和 LNM 之间的串扰机制仍然很大程度上未知。

目标

本研究旨在为 CC 患者构建一个 miRNA 依赖性预后模型，并通过调节 LDs 积累来研究 miR-532-5p 是否对 LNM 具有生物学影响。

方法

应用 LASSO-Cox 回归建立预后预测模型。miR-532-5p 在 RNA 表达 (P < 0.001) 和预后预测 (P < 0.0001) 方面的 P 值最低，因此被选作进一步研究。研究了预后 miR-532-5p 相关的竞争性内源性 RNA (ceRNA) 网络的功能作用，以阐明 LD 和 LNM 之间的串扰。使用定点诱变、双荧光素酶报告基因测定、RNA 免疫沉淀测定和救援实验确定了潜在机制。建立异种移植LNM模型以评估miR-532-5p和奥利司他联合治疗对肿瘤生长和LNM的影响。

结果

构建了一种新的 5-miRNA 预后特征，以更好地预测 CC 患者的预后。进一步的研究表明，miR-532-5p 通过调节 LDs 的积累来抑制上皮间质转化和淋巴管生成。有趣的是，我们还发现 LDs 积累促进了体外细胞转移。从机制上讲，我们展示了一个与 miR-532-5p 相关的 ceRNA 网络，其中 LINC01410 直接与 miR-532-5p 结合，并作为 miR-532-5p 海绵有效地发挥作用，以去抑制其靶基因脂肪酸合酶 ( FASN )。与 miR-532-5p 和 FASN 抑制剂-奥利司他联合治疗可进一步抑制体内肿瘤生长和 LNM。

结论

我们的研究结果强调了 miR-532-5p 调节的 LNM 的 LD 积累依赖性机制，并支持使用 miR-532-5p/orlistat 作为治疗 CC 中 LNM 患者的新策略。