Long non-coding RNA PVT1 promotes tumor progression by regulating the miR-143/HK2 axis in gallbladder cancer

Abstract

Background The long non-coding RNA PVT1 (lncRNA PVT1) has been reported to act as an oncogenic regulator of several cancers. However, its expression and function in gallbladder cancer (GBC) remain largely unknown. Methods In situ hybridization (ISH) and quantitative real-time PCR (qPCR) were performed to detect the expression of PVT1 and miR-143 in GBC tissues and cell lines. Immunohistochemistry (IHC) assays were performed to assess the expression of the hexokinase 2 (HK2) protein. The relationships among PVT1, miR-143 and HK2 were evaluated using dual-luciferase reporter, RNA immunoprecipitation (RIP) and biotin pull-down assays. The biological functions of PVT1, miR-143 and HK2 in GBC cells were explored with cell counting kit 8 (CCK-8), 5-ethynyl-20-deoxyuridine (EdU), colony formation, transwell, wound healing and glucose metabolism assays in vitro. For in vivo experiments, a xenograft model was used to investigate the effects of PVT1 and HK2 on GBC. Results PVT1 was upregulated in GBC tissues and cells and was positively associated with malignancies and worse overall survival. PVT1 knockdown inhibited cell proliferation, migration, and invasion in vitro and restrained tumor growth in vivo. Further studies demonstrated that PVT1 positively regulated HK2 expression via its competing endogenous RNA (ceRNA) activity on miR-143. Additionally, HK2 expression and function were positively correlated with PVT1. Furthermore, we observed that the PVT1/miR-143/HK2 axis promoted cell proliferation and metastasis by regulating aerobic glucose metabolism in GBC cells. Conclusions The results of our study reveal a potential ceRNA regulatory pathway in which PVT1 modulates HK2 expression by competitively binding to endogenous miR-143 in GBC cells, which may provide new insights into novel molecular therapeutic targets for GBC. Electronic supplementary material The online version of this article (10.1186/s12943-019-0947-9) contains supplementary material, which is available to authorized users.

TGF‐β‐activated lncRNA LINC00115 is a critical regulator of glioma stem‐like cell tumorigenicity

“…For example, the lncRNA activated by TGF-b (lncRNA-ATB) upregulated EMT factors by competitively binding the miR-200 family in hepatocellular carcinoma invasion and metastasis [27]. PVT1 played as a miR-143 sponge and upregulated hexokinase 2 (HK2) expression levels in gallbladder cancer [13]. In this study, we demonstrated LINC00115 as a new miRNA sponge of miR-200 family to promote ZEB1 signaling in GBM.…”

“…High TGF‐β activity confers poor prognosis in glioma patients . TGF‐β1 promotes the self‐renewal of glioma‐initiating cells (GICs) through induction of LIF or Sox2 , and inhibition of TGF‐β activity decreases the CD44 high /ID1 high GIC population through inhibition of ID1 and ID3 .…”

“…High TGF-b activity confers poor prognosis in glioma patients [12]. TGF-b1 promotes the self-renewal of glioma-initiating cells (GICs) through induction of LIF or Sox2 [13,14], and inhibition of TGF-b activity decreases the CD44 high / ID1 high GIC population through inhibition of ID1 and ID3 [11]. microRNAs (miRNAs) are small (20-22 nucleotides long) noncoding RNAs (ncRNAs) that have been recognized to play important roles in development and cancer [15].…”

MiR‐26b suppresses hepatocellular carcinoma development by negatively regulating ZNRD1 and Wnt/β‐catenin signaling

“…Immunohistochemical staining was performed was performed according to previously described method . In brief, the TMA slides were deparaffinised through xylene, antigen retrieved and blocked with bovine serum albumin.…”

“…Immunohistochemical staining was performed was performed according to previously described method. 22 In brief, the TMA slides were deparaffinised through xylene, antigen retrieved and blocked with bovine serum albumin. Subsequently, the slides were incubated in ZNRD1 antibody diluted 1:200 at 4°C overnight and carried out using the EnVision™ system (DAKO) according to the manufacturer's instructions.…”