Wang Ji, Qi Xinyue, Zhao Wangqian, Hao Ziyan, Wang Kehan, Li Yuting, Wang Yue, Zhang Yunshan
Department of Oncology, The Second Affiliated Hospital of Soochow University, Suzhou, P.R. China.
Cancer institute, Suzhou Medical College, Soochow university, Suzhou, P.R. China.
PLoS One. 2025 Aug 4;20(8):e0329847. doi: 10.1371/journal.pone.0329847. eCollection 2025.
Intrahepatic cholangiocarcinoma (iCCA) is a malignancy with difficult treatment and poor prognosis, whose pathogenesis could be associated with the expression patterns of circular RNAs (circRNAs). Here, we aim to investigate the effects and mechanism of hsa_circ_0006834 on iCCA proliferation. At first, hsa_circ_0006834 was proved to suppress iCCA cells proliferation and induce autophagy following the construction of hsa_circ_0006834 vector. And hsa_circ_0006834 is negatively linked with the proliferation, migration and invasion of iCCA cells through autophagy. Subsequently, RNA pull-down and dual-luciferase reporter assays were performed to validate the hsa_circ_0006834-has-miR-637-NGFR regulatory network. It was demonstrated by qPCR and Western blot that hsa_circ_0006834 stimulates the AMPK-mTOR pathway and triggers autophagy via NGFR after si-NGFR was synthesized and transfected into iCCA cells. Ultimately, after the successful knockdown of AMPK, the role of the AMPK-mTOR pathway in hsa_circ_0006834 regulating autophagy and proliferation was verified. Taken together, our findings revealed that hsa_circ_0006834 activates the AMPK-mTOR pathway and autophagy to suppress iCCA proliferation by has-miR-637-NGFR network, indicating the potential of hsa_circ_0006834 as a biomarker for iCCA diagnosis and therapy.
肝内胆管癌（iCCA）是一种治疗困难且预后较差的恶性肿瘤，其发病机制可能与环状RNA（circRNA）的表达模式有关。在此，我们旨在研究hsa_circ_0006834对iCCA增殖的影响及机制。首先，在构建hsa_circ_0006834载体后，证实hsa_circ_0006834可抑制iCCA细胞增殖并诱导自噬。并且hsa_circ_0006834通过自噬与iCCA细胞的增殖、迁移和侵袭呈负相关。随后，进行RNA下拉和双荧光素酶报告基因检测以验证hsa_circ_0006834-has-miR-637-NGFR调控网络。在合成si-NGFR并转染至iCCA细胞后，通过qPCR和蛋白质印迹法证明hsa_circ_0006834通过NGFR刺激AMPK-mTOR通路并触发自噬。最终，在成功敲低AMPK后，验证了AMPK-mTOR通路在hsa_circ_0006834调节自噬和增殖中的作用。综上所述，我们的研究结果表明，hsa_circ_0006834通过has-miR-637-NGFR网络激活AMPK-mTOR通路和自噬以抑制iCCA增殖，表明hsa_circ_0006834作为iCCA诊断和治疗生物标志物的潜力。
