Jing Zhao1,2, Jing Wu3, Yunyun Qin1, Wenhong Zhang4, Guangjian Huang5, Lunxiu Qin6,7. 1. Department of General Surgery, Huashan Hospital, Fudan University, Shanghai, 200040, People's Republic of China. 2. Cancer Metastasis Institute, Fudan University, Shanghai, 200040, People's Republic of China. 3. Department of Infectious Diseases, Huashan Hospital, Fudan University, Shanghai, 200040, People's Republic of China. 4. Department of Infectious Diseases, Huashan Hospital, Fudan University, Shanghai, 200040, People's Republic of China. zhangwenhong@fudan.edu.cn. 5. Department of General Surgery, Huashan Hospital, Fudan University, Shanghai, 200040, People's Republic of China. gjhuang@fudan.edu.cn. 6. Department of General Surgery, Huashan Hospital, Fudan University, Shanghai, 200040, People's Republic of China. qinlx@fudan.edu.cn. 7. Cancer Metastasis Institute, Fudan University, Shanghai, 200040, People's Republic of China. qinlx@fudan.edu.cn.
Abstract
PURPOSE: Vasculogenic mimicry (VM), a vessel-like network formed by highly aggressive tumor cells, plays an important role in accelerating cancer progression. This special vascularization pattern is closely associated with a poor prognosis in various cancers. As yet, however, the regulatory mechanism of VM formation is largely unknown. In this study, we assess whether the long noncoding RNA PVT1 is involved in VM generation in gastric cancer. METHODS: VM formation was determined by immunohistochemistry using PAS/CD31 double staining in gastric cancers and Matrigel tube formation in vitro. qRT-PCR and Western blotting were used to assess mRNA and protein expression. Interaction between PVT1 and STAT3 was determined using a RNA pull-down assay. Luciferase reporter and chromatin immunoprecipitation assays were performed to evaluate transcriptional activity of STAT3 on the Slug gene promoter. RESULTS: We found that PVT1 can induce VM generation both in vitro and in vivo. Mechanistically, we found that PVT1 interacted with and activated STAT3 through a 850-1770 nt fragment. PVT1 facilitated STAT3 recruitment to the Slug promoter and transcriptionally enhanced Slug expression, thereby triggering epithelial-to-mesenchymal transition (EMT) and VM capillary formation. STAT3 inhibition effectively blocked PVT1-mediated VM. In primary gastric cancer samples, a positive correlation was found between PVT1 and Slug upregulation, and patients with a high PVT1 and Slug expression exhibited markedly shorter survival times. CONCLUSION: Our results shed light on the role of PVT1 in gastric cancer cell-dependent VM formation. Our findings provide valuable clues for the design of new anti-angiogenic therapeutic strategies. The PVT1/STAT3 axis may serve as a potential target in gastric cancer treatment.
PURPOSE: Vasculogenic mimicry (VM), a vessel-like network formed by highly aggressive tumor cells, plays an important role in accelerating cancer progression. This special vascularization pattern is closely associated with a poor prognosis in various cancers. As yet, however, the regulatory mechanism of VM formation is largely unknown. In this study, we assess whether the long noncoding RNA PVT1 is involved in VM generation in gastric cancer. METHODS: VM formation was determined by immunohistochemistry using PAS/CD31 double staining in gastric cancers and Matrigel tube formation in vitro. qRT-PCR and Western blotting were used to assess mRNA and protein expression. Interaction between PVT1 and STAT3 was determined using a RNA pull-down assay. Luciferase reporter and chromatin immunoprecipitation assays were performed to evaluate transcriptional activity of STAT3 on the Slug gene promoter. RESULTS: We found that PVT1 can induce VM generation both in vitro and in vivo. Mechanistically, we found that PVT1 interacted with and activated STAT3 through a 850-1770 nt fragment. PVT1 facilitated STAT3 recruitment to the Slug promoter and transcriptionally enhanced Slug expression, thereby triggering epithelial-to-mesenchymal transition (EMT) and VM capillary formation. STAT3 inhibition effectively blocked PVT1-mediated VM. In primary gastric cancer samples, a positive correlation was found between PVT1 and Slug upregulation, and patients with a high PVT1 and Slug expression exhibited markedly shorter survival times. CONCLUSION: Our results shed light on the role of PVT1 in gastric cancer cell-dependent VM formation. Our findings provide valuable clues for the design of new anti-angiogenic therapeutic strategies. The PVT1/STAT3 axis may serve as a potential target in gastric cancer treatment.
Authors: Hashem Khanbabaei; Saeedeh Ebrahimi; Juan Luis García-Rodríguez; Zahra Ghasemi; Hossein Pourghadamyari; Milad Mohammadi; Lasse Sommer Kristensen Journal: J Exp Clin Cancer Res Date: 2022-09-16