Long noncoding RNA PANDA promotes esophageal squamous carcinoma cell progress by dissociating from NF-YA but interact with SAFA

https://doi.org/10.1016/j.prp.2019.152604Get rights and content

Abstract

Esophageal squamous cell carcinoma (ESCC) is one of the major global health problems, especially in Asia. Long non-coding RNAs (lncRNAs) have been increasingly identified and characterized in almost every aspect of biology, especially in cancer biology. This research desires to explore the regulatory mechanism of lncRNA PANDA (PANDA) on ESCC process. Quantitative real-time PCR (qRT-PCR) was carried out to detect the PANDA expression, which was up-regulated in matched cancerous tissues and adjacent noncancerous tissues from 134 patients and 9 ESCC cell lines. Higher expression of PANDA in ESCC tissues was associated with TNM stage, advanced clinical stage, and shorter overall survival of ESCC patients by MTT, EDU, colony formation assay and flow cytometry in KYSE180 and KYSE450 cells. Exogenous down-regulation of PANDA expression significantly suppressed ESCC cells proliferation and colony formation by arresting G1-S checkpoint transition in vitro, and retarded the development of tumors in vivo. Meanwhile, qRT-PCR and western blot assays showed that depletion of PANDA reduced E2F1, cyclinD1, cyclinD2, cyclinE1 and Bcl-2 expression. RIP showed the interaction between PANDA and NF-YA or SAFA. Our findings suggested that, PANDA drifted away from NF-YA to promote the expression of NF-YA-E2F1 co-regulated proliferation-promoting genes, and to limit the cell apoptosis. In addition, PANDA binds SAFA to switch on the tumor proliferation program through CyclinD1/2-Cyclin E1 and Bcl-2 pathways. PANDA could serve as a potential prognostic biomarker and therapeutic target for ESCC.

Introduction

Esophageal cancer (EC) is the eighth most common malignant tumor worldwide and the sixth most common cause of death from cancer [1]. Esophageal cancer, derived from epithelia, consists of two subtypes: esophageal squamous cell carcinoma (ESCC) and esophageal adenocarcinoma, which have different etiologic and pathologic characteristics [2]. In Asia, the predominant pathological type is ESCC, which is often diagnosed at the advanced stage, and China is one of the high-risk esophageal cancer areas [3]. To date, the prognosis for ESCC patients is not well improved with a rate of less than 10% 5-year survival [4]. Therefore, it is urgent to understand the detailed interactions and regulatory mechanisms of key pathways involved in the tumor-genesis and progression of ESCC and find molecular markers for early detection and diagnosis.

Recent studies have revealed that epigenetic regulation also participates in cancer development and progression [5]. Long noncoding RNAs (LncRNAs), which occupy the majority of human genome, have been shown to play an important role in the regulation of gene transcription, translation and widespread regulators involved in cell proliferation, migration and apoptosis [[6], [7], [8], [9], [10]]. Mounting evidence indicate that lncRNAs expression is misregulated and contributes to the development and progression of multiple cancers. Their dysregulation has been found in various types of carcinomas, including breast cancer, colon cancer, hepatocellular carcinoma and lung cancer. LncRNAs are often up-regulated and down-regulated, and may serve as oncogenes or tumor suppressors in cancers [[11], [12], [13]]. However, the roles of lncRNAs in ESCC are still not well documented and needed to be further explored.

LncRNA PANDA (PANDA), a 5-capped and polyadenylated non-spliced lncRNA, is transcripted from approximately 5 kb upstream of the CDKN1A transcriptional start site. PANDA expression could be induced by DNA damage and limit expression of pro-apoptotic genes by interacting with the transcription factor NF-YA in a p53-dependent manner [14]. Moreover, PANDA deletion could increase human fibroblasts cells sensitivity to doxorubicin and induce cell apoptosis. However, its expression and biological function in ESCC remain poorly understood. In this study, the expression pattern and biological functions of PANDA in ESCC development were explored. The correlations between PANDA and the clinical outcomes of ESCC patients were evaluated, and the effects of PANDA on proliferation and apoptosis of ESCC cells were also investigated both in vitro and in vivo. Moreover, knockdown of PANDA could affect multiple gene expression involved in regulating phase checkpoints of cell cycle, such as E2F1. This study might provide a novel mechanism and potential therapeutic target for ESCC.

Section snippets

Tissue sample collection and patient data

All samples were collected from informed consent individuals according to protocols approved by the ethics committee of Nanjing Hospital affiliated with Nanjing Medical University. A total of 134 paired samples of ESCC (TNM Stage 0 to IV), including cancer tissues and corresponding adjacent non-cancer tissues were obtained from patients who underwent surgery in our hospital from 2005 to 2012. Only 76 cases of 134 patients had follow-up materials of 5 yrs used for survival analysis for 5 yrs. No

LncRNA PANDA is up-regulated in of ESCC tissues

To validate levels of PANDA expression, we conducted qRT-PCR analysis to measure the PANDA expression in 134 paired cancerous and adjacent noncancerous tissues of ESCC. PANDA expression was up-regulated in 67.5% ESCC sample (91 of 134 cases, p < 0.05) 91 cancerous tissues and down-regulated in 43 cancerous tissues compared with their paired adjacent noncancerous tissues (Fig. 1A and 1B). All ESCC sample were classified into two groups based on the median value of relative PANDA expression. The

Discussion

The human genome contains a large number of lncRNAs that are dynamically expressed in a tissue-, differentiation-, cell type- or developmental stage-specific manner, indicating specific functions of lncRNAs in the development of diseases [[25], [26], [27]]. The cellular functions of most recently discovered lncRNAs then needed to be elucidated. For each individual molecule, it needs to be established whether it executes important functions or just represents “transcriptional noise” or

Conclusions

In this study, we figured out the lncRNA PANDA who could promote the ESCC progression in vitro and in vivo. We reported that PANDA drifted away from NF-YA to promote the expression of NF-YA-E2F1 coregulated proliferation-promoting genes, and to limit the ESCC cell apoptosis. In addition, PANDA binding SAFA to switch on the tumor proliferation programme though CyclinD1/2-Cyclin E1 and Bcl-2 pathways in ESCC.

Availability of data and materials

All data generated or analyzed during this study are included in this published article.

Funding

This work was supported by Suzhou Science and Technology Development Project (SNG201607, SNG2017049), and 2018 Municipal Industrial Development Guidance Fund (Technology Innovation Special Project) (s201808).

Declaration of Competing Interest

The authors declare that they have no competing interests

References (58)

  • A. Jemal et al.

    Global Cancer statistics

    CA Cancer J. Clin.

    (2011)
  • K. Matsushima et al.

    Mirna-205 modulates cellular invasion and migration via regulating zinc finger E-Box binding homeobox 2 expression in esophageal squamous cell carcinoma cells

    J. Transl. Med.

    (2011)
  • M. Hongo et al.

    Epidemiology of reflux disease and cle in East Asia

    J. Gastroenterol.

    (2003)
  • L.A. Torre et al.

    Global Cancer statistics, 2012

    CA Cancer J. Clin.

    (2015)
  • E.A. Dethoff et al.

    Functional complexity and regulation through Rna dynamics

    Nature

    (2012)
  • C.T. Ong et al.

    Enhancer function: new insights into the regulation of tissue-specific gene expression

    Nat. Rev. Genet.

    (2011)
  • M.C. Tsai et al.

    Long noncoding Rna as modular scaffold of histone modification complexes

    Science

    (2010)
  • M.C. Tsai et al.

    Long intergenic noncoding Rnas: new links in cancer progression

    Cancer Res.

    (2011)
  • W.C. Liang et al.

    Lncrna-nef antagonized epithelial to mesenchymal transition and cancer metastasis via cis-regulating Foxa2 and inactivating Wnt/Beta-Catenin signaling

    Oncogene

    (2018)
  • W. Su et al.

    Silencing of long noncoding Rna Mir22hg triggers cell Survival/Death signaling via oncogenes Ybx1, met, and P21 in lung cancer

    Cancer Res.

    (2018)
  • Z. Xing et al.

    Expression of long noncoding Rna Yiya promotes glycolysis in breast cancer

    Cancer Res.

    (2018)
  • T. Hung et al.

    Extensive and coordinated transcription of noncoding rnas within cell-cycle promoters

    Nat. Genet.

    (2011)
  • A.C. Tsamandas et al.

    The potential role of Bcl-2 expression, apoptosis and cell proliferation (Ki-67 expression) in cases of gastric carcinoma and correlation with classic prognostic factors and patient outcome

    Anticancer Res.

    (2009)
  • Q. Ke et al.

    Impairment of liver regeneration by the histone deacetylase inhibitor valproic acid in mice

    J. Zhejiang Univ. Sci. B

    (2012)
  • Y.P. Tao et al.

    Associations between polymorphisms in Il-12a, Il-12b, Il-12rbeta1, Il-27 gene and serum levels of Il-12p40, Il-27p28 with esophageal cancer

    J. Cancer Res. Clin. Oncol.

    (2012)
  • J.G. Wu et al.

    Expressions and clinical significances of C-Met, P-Met and E2f-1 in human gastric carcinoma

    BMC Res. Notes

    (2014)
  • Y. Zhang et al.

    Reciprocal activation between Plk1 and Stat3 contributes to survival and proliferation of esophageal cancer cells

    Gastroenterology

    (2012)
  • T. Hung et al.

    Extensive and coordinated transcription of noncoding Rnas within cell-cycle promoters

    Nat. Genet.

    (2013)
  • R. Helbig et al.

    Scaffold attachment factor a (Saf-a) is concentrated in inactive X chromosome territories through its Rgg domain

    Chromosoma

    (2003)
  • Cited by (12)

    • Roles of lncRNAs in the transcription regulation of HIV-1

      2022, Biomedical Journal
      Citation Excerpt :

      To achieve integration, the IN cleaves the host cell's genomic DNA to produce double-strand breaks. Two unique p53-dependent lncRNAs, long intergenic noncoding RNA-p21 (lincRNA-p21) and PANDA (p21-related noncoding RNA DNA damage activation), are derived from the gene encoding the critical cell cycle regulator cyclin dependent kinase inhibitor 1A (CDKN1A) (also known asp21) promoter, a canonical transcriptional target of p53 [63,64,67,75], and have been identified in two published studies by comparing differentially expressed lncRNAs after HIV infection. Since p53-binding proteins play a crucial role in the cell response to the breakage of double-stranded DNA in the genome, it is not surprising that p53-dependent lncRNAs are differentially expressed during viral infection [65,66].

    • The lncRNA APOLO interacts with the transcription factor WRKY42 to trigger root hair cell expansion in response to cold

      2021, Molecular Plant
      Citation Excerpt :

      PANDA is transcribed from the promoter region of the CDKN1A gene and interacts with the TF NF-YA to limit the expression of pro-apoptotic genes. The activity of PANDA has been linked to the progression of different tumors (Kotake et al., 2016; Shi et al., 2019). Interestingly, it was shown that, in addition to NF-YA, PANDA interacts with the scaffold attachment factor A (SAFA) as well as PRC1 and PRC2 to modulate cell senescence.

    View all citing articles on Scopus
    1

    Contributed to the work equally.

    View full text