Background
To maintain the urgent demand for tumorigenesis and aggressiveness, pancreatic cancer cells have extensively reprogrammed energy metabolism [
1]. One of the most common and important alterations of metabolism is aerobic glycolysis, which is termed the “Warburg effect”. Glycolysis displays a less efficient metabolism with more lactate production, lower extracellular pH value, and more glucose consumption, even under an oxygen-enriched microenvironment [
2]. Pancreatic cancer cells show significantly activated glycolysis, which was associated with poor prognosis in pancreatic cancer (PC) patients [
3]. Thereby, targeting the Warburg effect has been regarded as a promising therapeutic direction in tumors. Recent studies revealed that disrupting SLC2A1, LDHA or other key glycolytic enzymes resulted in remarkably reduced conversion of glucose to pyruvate and ATP, and elevated mitochondrial oxidative phosphorylation [
4]. Meanwhile, inhibitors of SLC2A1 or LDHA in combination with tumor chemotherapeutics displayed significant synergistic antitumor effect in vitro, which highlighting the promising role for cancer therapeutic strategy of glycolysis [
5,
6]. Nevertheless, the metabolic process and molecular mechanisms of glycolysis in PC progression remain uncertain and need to be further determined.
Long noncoding RNAs (lncRNAs) are a type of special transcript longer than 200 nucleotides and lack protein-coding potential. Although lncRNAs were regarded as merely transcriptional ‘‘noise’’ before, emerging evidence has shown that lncRNAs play pivotal roles in transcriptional regulation, epigenetic gene regulation, and tumors [
7]. And our previous study has revealed that lncRNA GLS-AS, dysregulated under glucose and glutamine deprivation, repressed pancreatic cancer progression by repressing the Myc/GLS pathway [
8]. Specifically, emerging studies have shown the key role of lncRNAs in the regulation of glycolysis in cancers. For example, lncRNA MACC1-AS1 promotes glycolysis progression and anti-oxidative capabilities under metabolic stress in gastric cancer [
9]. Besides, lncRNA LINRIS has been reported as an independent prognostic factor that participated in the regulation of aerobic glycolysis of colorectal cancer [
10]. Similarly, Wang et al. have reported that lncRNA PVT1 facilitates PC glycolysis and progression by regulating the miR-519d-3p/HIF-1A pathway [
11]. Nevertheless, the molecular mechanisms of lncRNAs for the Warburg effect and its role in PC progression have not been fully discovered. Therefore, our present study sought to find a lncRNA that is critical for glycolysis and tumorigenesis of PC.
In this study, we identified that DICER1-AS1 was down-regulated in PC and was correlated with a poor prognosis. Meanwhile, both in vitro and in vivo experiments validated that DICER1-AS1 functioned as an inhibitor of PC in glycolysis and tumorigenesis by promoting DICER1-mediated maturation of miR-5586-5p. Notably, we further disclosed DICER1-AS1 was degraded in glucose deprivation and further explored the m6A-correlated regulation.
Discussion
Recently, the non-coding RNAs (ncRNAs), including miRNAs and lncRNAs have received growing attention in human tumors [
26], particularly in pancreatic cancer [
8,
27,
28]. Although studies have evaluated the involvement of lncRNAs in the dysregulation of glycolysis metabolism in pancreatic cancers, the precise mechanism needs to be further investigated [
29,
30]. In this study, we identify that DICER1-AS1 is an inhibitor for glycolysis and tumorigenesis of pancreatic cancer by inducing miRNA-mediated mRNA degradation of glycolytic genes. Moreover, the present study indicates that DICER1-AS1 is epigenetically downregulated by its target m6A reader YTHDF3, which forms a negative feedback.
The bioinformatic analyses showed that DICER1-AS1 was downregulated in PC tissues and negatively correlated with glycolytic pathway and overall survival. We further identified that overexpression of DICER1-AS1 significantly inhibited glycolysis and consequently prohibited proliferation and metastasis of pancreatic cancer cells both in vitro and in vivo. Moreover, we further displayed that DICER1-AS1 was downregulated in PC tissues in our cohort 1 and associated with poor overall survival of PC patients. Thus, our study implicates that DICER1-AS1 is an inhibitor for glycolysis and tumorigenesis of pancreatic cancer. Differently, research from Gu et al. reported that DICER1-AS1 promoted the proliferation, invasion, and autophagy of osteosarcoma cells via miR-30b/ATG5 axis [
31]. Other research also indicated that DICER1-AS1 functioned as promoter in colorectal cancer by regulating miR-296-5p/STAT3 pathway [
32]. Therefore, these researches implicate that the function of DICER1-AS1 in heterogeneous cancer, which needs further investigation.
Natural antisense transcripts are a cluster of RNAs that are transcribed from the opposite strand of a sense DNA strand. It’s reported that antisense RNA can alter the expression of sense genes at the transcriptional or post-transcriptional level [
33,
34]. Similarly, our present study demonstrated the inhibition on glycolysis of DICER1-AS1 was dependent on regulating DICER1 transcription. Meanwhile, as a critical processor for maturation of miRNA, we further founded that DICER1 promoted maturation of miR-5586-5p. Research also showed that knockdown of DICER1 significantly inhibited xenograft tumor growth of BxPC-3 cells by suppressing expression of miR-22, miR-143, let-7i, and miR-29b [
35]. PICK1 inhibits the processing of pre-mir-615-3p to mature miR-615-3p via interfering with the binding of DICER1 to Smad2/3 in breast cancer cells [
36]. Thereby, our research provides a new clue for interaction between lncRNA and miRNA which involving DICER1-mediated miRNA maturation.
YY1 is a ubiquitously distributed transcription factor belonging to the GLI-Krüppel family of zinc-finger protein, associated with multiple cellular processes [
37]. YY1 suppressed the proliferation via inhibiting the transcription of lncRNA SOX2OT, thereby decreased the expression of its target gene SOX2 in PC [
38]. Other studies also showed that YY1 suppressed the proliferation, invasion, and metastatic properties of PC cells by downregulating MMP10 [
39], as well as FER and MMP2 [
40]. Similarly, our present study validated that DICER1-AS1 promoted DCIER1 transcription by recruiting YY1 to promoter of DICER1. Meanwhile, YY1 was downregulated in PC tissues and correlated with DICER1 expression and poor prognosis of PC patients. Therefore, these data also identify that YY1 functions as an inhibitor in PC by interacting with DICER1-AS1/DICER1 pathway.
It’s reported that miRNAs play a vital role in metabolic homeostasis including glycolysis in tumors. For example, circMAT2B upregulated PKM2 by functioning as a “sponge” of miR-338-3p, meanwhile promotes the glycolysis level and hepatocellular carcinoma progression [
41]. Besides, cell-secreted miR-122 is associated with metastasis in breast cancer patients by inhibiting glycolysis-related pyruvate kinase [
42]. The present study demonstrated that miR-5586-5p induced degradation of glycolytic genes including SLC2A1, LDHA, HK2, and PGK1 by binding to 3’UTR of each mRNA, which consequently inhibited glycolysis of PC cells. Meanwhile, GEO database displayed that the 4 glycolytic genes were highly expressed in PC tissues and negatively correlated with miR-5586-5p and overall survival of PC patients. Similarly, research from Bai et al. showed that a seven-miRNA expression-based prognostic signature including miR-5586-5p was downregulated in PC which might stratify patients with PC into low- and high-risk groups [
43]. Thereby, these data intensively reminder that miR-5586 is an inhibitor of PC and provides further evidence that miRNAs play critical roles in glycolysis of tumor.
Recent studies showed that m6A modification, as the most abundant RNA modification, was responsible for the dysregulation of lncRNAs [
44]. Previous studies showed that the stability of lncRNA GAS5 was destroyed by YTHDF3 and consequently promoted progression of colorectal cancer [
24]. Similarly, our study showed glucose deprivation significantly induced degradation of DICER1-AS1, which was mediated by m6A reader YTHDF3. Interestingly, our research also displayed that YTHDF3 was a critical target for miR-5586-5p. To explore the comprehensive effects of YTHDF3/DICER1-AS1/DICER1/miR-5586-5p axis on the glycolysis and prognosis of PC, we performed IHC assay and Kaplan–Meier survival analyses. The results demonstrated that high expression of DICER1-AS1 correlated with high expression of DCIER1, but correlated with low expression of YTHDF3 and the four glycolytic genes. Moreover, high expression of DICER1-AS1 along with high expression of DICER1 or miR-5586 was associated with longer overall survival. On the contrary, high expression of YTHDF3 along with low expression of DICER1 or DICER1-AS1 demonstrated shorter overall survival. Moreover, the in vivo experiments further identified that overexpression of DICER1-AS1 significantly inhibited the growth and metastasis of PC tumors.
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