Zhu Mengjing, Zhou Ling, Fu Jian, Wang Yijin, Xu Xiaofeng, Wu Jun, Kong Xiangyi, Li Jian, Zhou Zhe, Zhou Huaijun
Department of Gynecology, Nanjing Drum Tower Hospital, The Affiliated Hospital of Nanjing University Medical School, Nanjing 210008, China.
Department of Gynecology, Suqian People's Hospital of Nanjing Drum Tower Hospital Group, Suqian 223800, China.
J Oncol. 2022 Nov 26;2022:3332485. doi: 10.1155/2022/3332485. eCollection 2022.
The neurotrophic factor Artemin (ARTN) is involved in tumor proliferation and metastasis. Nonetheless, ARTN's significance in cervical cancer (CC) has not been studied. In our study, we propose to investigate the biological function of ARTN in CC as well as its particular regulatory mechanism.
Immunohistochemistry (IHC) was used to examine the degree of ARTN protein expression in CC patient tissue. Real-time PCR and Western blotting were performed to reveal related genes' levels in CC cells. The CCK-8 test, the colony formation assay, the wound-healing assay, and the transwell assay were utilized to determine the proliferation, migration, and invasion capabilities, respectively. To generate lung metastasis models, stable ARTN-expressing SiHa cells were injected into the caudal tail vein of mice. IHC was used to examine the protein levels in CC mice model tissues.
ARTN was overexpressed in CC tissues relative to normal cervical tissues and linked positively with lymph node metastases (=0.012) and recurrence (=0.015) in CC patients. In vitro, ARTN overexpression promoted the proliferation, invasion, and migration of CC cells. In contrast, the consequences of depleting endogenous ARTN were the opposite. Moreover, overexpression of ARTN increased lung metastasis of CC cells in vivo and shortened the lifespan of mice models. In addition, ARTN overexpression significantly enhanced AKT phosphorylation on Ser473 and mTOR phosphorylation on Ser2448 and promoted the epithelial-mesenchymal transition (EMT) cascade. In addition, rapamycin, a selective inhibitor of mTORC1, might rescue the EMT phenotype caused by ARTN.
Our findings suggested that ARTN may enhance CC metastasis through the AKT/mTORC1 pathway. ARTN is anticipated to be a novel potential therapeutic target for the treatment of CC metastases.
神经营养因子Artemin（ARTN）参与肿瘤增殖和转移。然而，ARTN在宫颈癌（CC）中的意义尚未得到研究。在我们的研究中，我们提议研究ARTN在CC中的生物学功能及其特定的调控机制。
采用免疫组织化学（IHC）检测CC患者组织中ARTN蛋白的表达程度。进行实时PCR和蛋白质印迹法以揭示CC细胞中相关基因的水平。分别利用CCK-8试验、集落形成试验、伤口愈合试验和Transwell试验来测定增殖、迁移和侵袭能力。为了建立肺转移模型，将稳定表达ARTN的SiHa细胞注入小鼠尾静脉。采用IHC检测CC小鼠模型组织中的蛋白水平。
相对于正常宫颈组织，ARTN在CC组织中过表达，并且与CC患者的淋巴结转移（P=0.012）和复发（P=0.015）呈正相关。在体外，ARTN过表达促进了CC细胞的增殖、侵袭和迁移。相反，耗尽内源性ARTN的结果则相反。此外，ARTN过表达增加了CC细胞在体内的肺转移，并缩短了小鼠模型的寿命。此外，ARTN过表达显著增强了Ser473位点的AKT磷酸化和Ser2448位点的mTOR磷酸化，并促进了上皮-间质转化（EMT）级联反应。此外，mTORC1的选择性抑制剂雷帕霉素可能挽救由ARTN引起的EMT表型。
我们的研究结果表明，ARTN可能通过AKT/mTORC1途径增强CC转移。ARTN有望成为治疗CC转移的新型潜在治疗靶点。
