XIST Induced by JPX Suppresses Hepatocellular Carcinoma by Sponging miR-155-5p

Purpose The influence of X-inactive specific transcript (XIST) and X-chromosome inactivation associated long non-coding RNAs (lncRNAs) just proximal to XIST (JPX) on hepatocellular carcinoma (HCC) remains controversial in light of previous reports, which the present study aimed to verify. Materials and Methods The DIANA lncRNA-microRNA (miRNA) interaction database was used to explore miRNA interactions with JPX or XIST. JPX, XIST, and miR-155-5p expression levels in paired HCC specimens and adjacent normal tissue were analyzed by RT-qPCR. Interaction between XIST and miR-155-5p was verified by dual luciferase reporter assay. Expression levels of miR-155-5p and its known target genes, SOX6 and PTEN, were verified by RT-qPCR and Western blot in HepG2 cells with or without XIST knock-in. The potential suppressive role of XIST and JPX on HCC was verified by cell functional assays and tumor formation assay using a xenograft model. Results JPX and XIST expression was significantly decreased in HCC pathologic specimens, compared to adjacent tissue, which correlated with HCC progression and increased miR-155-5p expression. Dual luciferase reporter assay revealed XIST as a direct target of miR-155-5p. XIST knock-in significantly reduced miR-155-5p expression level and increased that of SOX6 and PTEN, while significantly inhibiting HepG2 cell growth in vitro, which was partially reversed by miR-155-5p mimic transfection. JPX knock-in significantly increased XIST expression and inhibited HepG2 cell growth in vitro or tumor formation in vivo in a XIST dependent manner. Conclusion JPX and XIST play a suppressive role in HCC. JPX increases expression levels of XIST in HCC cells, which suppresses HCC development by sponging the cancer promoting miR-155-5p.

[1]  Panayiotis Tsanakas,et al.  DIANA-LncBase v2: indexing microRNA targets on non-coding transcripts , 2015, Nucleic Acids Res..

[2]  Bo Wang,et al.  Role of microRNA‐155 at early stages of hepatocarcinogenesis induced by choline‐deficient and amino acid–defined diet in C57BL/6 mice , 2009, Hepatology.

[3]  Liqun Wu,et al.  X-inactive-specific transcript of peripheral blood cells is regulated by exosomal Jpx and acts as a biomarker for female patients with hepatocellular carcinoma , 2017, Therapeutic advances in medical oncology.

[4]  Yue Wang,et al.  Long noncoding RNA HULC modulates abnormal lipid metabolism in hepatoma cells through an miR-9-mediated RXRA signaling pathway. , 2015, Cancer research.

[5]  P. Pandolfi,et al.  The multilayered complexity of ceRNA crosstalk and competition , 2014, Nature.

[6]  Xuan Liang,et al.  MicroRNA‐155‐5p promotes hepatocellular carcinoma progression by suppressing PTEN through the PI3K/Akt pathway , 2017, Cancer science.

[7]  C. Rougeulle,et al.  Origin and evolution of the long non-coding genes in the X-inactivation center. , 2011, Biochimie.

[8]  Yuh-Shan Jou,et al.  Xist reduction in breast cancer upregulates AKT phosphorylation via HDAC3-mediated repression of PHLPP1 expression , 2016, Oncotarget.

[9]  Carolyn J. Brown,et al.  Characterization of expression at the human XIST locus in somatic, embryonal carcinoma, and transgenic cell lines. , 2003, Genomics.

[10]  G. Pintacuda,et al.  Function by Structure: Spotlights on Xist Long Non-coding RNA , 2017, Front. Mol. Biosci..

[11]  Shuhan Sun,et al.  Long noncoding RNA DANCR increases stemness features of hepatocellular carcinoma by derepression of CTNNB1 , 2016, Hepatology.

[12]  Lei Wang,et al.  LncRNA XIST acts as a tumor suppressor in prostate cancer through sponging miR-23a to modulate RKIP expression , 2017, Oncotarget.

[13]  Rui Zhang,et al.  Long non-coding RNA XIST regulates PDCD4 expression by interacting with miR-21-5p and inhibits osteosarcoma cell growth and metastasis , 2017, International journal of oncology.

[14]  Liuqing Yang,et al.  Long Noncoding RNA in Cancer: Wiring Signaling Circuitry. , 2017, Trends in cell biology.

[15]  Wei Wang,et al.  MicroRNA-155 promotes tumor growth of human hepatocellular carcinoma by targeting ARID2. , 2016, International journal of oncology.

[16]  F. Zhou,et al.  Downregulation of long non-coding RNAs JPX and XIST is associated with the prognosis of hepatocellular carcinoma. , 2017, Clinics and research in hepatology and gastroenterology.

[17]  L. Zhang,et al.  Long noncoding RNA FTX inhibits hepatocellular carcinoma proliferation and metastasis by binding MCM2 and miR-374a , 2016, Oncogene.

[18]  Eda Yildirim,et al.  Xist RNA Is a Potent Suppressor of Hematologic Cancer in Mice , 2013, Cell.

[19]  M. McCrae,et al.  Aberrant expression of microRNA 155 may accelerate cell proliferation by targeting sex‐determining region Y box 6 in hepatocellular carcinoma , 2012, Cancer.

[20]  A. Bhan,et al.  Long Noncoding RNA and Cancer: A New Paradigm. , 2017, Cancer research.

[21]  Zeyi Liu,et al.  Long non-coding RNA XIST promotes TGF-β-induced epithelial-mesenchymal transition by regulating miR-367/141-ZEB2 axis in non-small-cell lung cancer. , 2018, Cancer letters.

[22]  Jeannie T. Lee,et al.  Jpx RNA Activates Xist by Evicting CTCF , 2013, Cell.

[23]  J. Gribnau,et al.  X chromosome inactivation: silencing, topology and reactivation. , 2017, Current opinion in cell biology.

[24]  A. Lindblom,et al.  High incidence of skewed X chromosome inactivation in young patients with familial non-BRCA1/BRCA2 breast cancer , 2005, Journal of Medical Genetics.

[25]  Xianqun Fan,et al.  CANT1 lncRNA Triggers Efficient Therapeutic Efficacy by Correcting Aberrant lncing Cascade in Malignant Uveal Melanoma. , 2017, Molecular therapy : the journal of the American Society of Gene Therapy.

[26]  Yaoyong Lu,et al.  Long non-coding RNA XIST promotes cell growth by regulating miR-139-5p/PDK1/AKT axis in hepatocellular carcinoma , 2017, Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine.

[27]  miR-155-5p modulates malignant behaviors of hepatocellular carcinoma by directly targeting CTHRC1 and indirectly regulating GSK-3β-involved Wnt/β-catenin signaling , 2017, Cancer Cell International.

[28]  B. Shen,et al.  The role of mmu‐miR‐155‐5p‐NF‐κB signaling in the education of bone marrow‐derived mesenchymal stem cells by gastric cancer cells , 2018, Cancer medicine.

[29]  M. Waye,et al.  Hotair mediates hepatocarcinogenesis through suppressing miRNA-218 expression and activating P14 and P16 signaling. , 2015, Journal of hepatology.

[30]  Jeannie T. Lee,et al.  The Long Noncoding RNA, Jpx, Is a Molecular Switch for X Chromosome Inactivation , 2010, Cell.

[31]  I. Ng,et al.  Non-coding RNAs in hepatocellular carcinoma: molecular functions and pathological implications , 2018, Nature Reviews Gastroenterology & Hepatology.

[32]  Xiaoyang Wang,et al.  Long non-coding RNA XIST regulates PTEN expression by sponging miR-181a and promotes hepatocellular carcinoma progression , 2017, BMC Cancer.

[33]  Yuehua Jiang,et al.  Prognostic value of microRNAs in hepatocellular carcinoma: a meta-analysis , 2017, Oncotarget.

[34]  Pier Paolo Pandolfi,et al.  ceRNA cross-talk in cancer: when ce-bling rivalries go awry. , 2013, Cancer discovery.

[35]  Wenbo Li,et al.  Knockdown of long non-coding RNA XIST inhibits cell viability and invasion by regulating miR-137/PXN axis in non-small cell lung cancer. , 2018, International journal of biological macromolecules.

[36]  D. Hua,et al.  Long Noncoding RNA XIST Regulates miR-137-EZH2 Axis to Promote Tumor Metastasis in Colorectal Cancer. , 2018, Oncology research.

[37]  C. Deng,et al.  The XIST Noncoding RNA Functions Independently of BRCA1 in X Inactivation , 2007, Cell.

[38]  P. Radice,et al.  X chromosome inactivation pattern in BRCA gene mutation carriers. , 2013, European journal of cancer.

[39]  Y. Yang,et al.  MicroRNA-92b promotes hepatocellular carcinoma progression by targeting Smad7 and is mediated by long non-coding RNA XIST , 2016, Cell Death and Disease.

[40]  Lei Shi,et al.  Hepatocellular carcinoma‐associated mesenchymal stem cells promote hepatocarcinoma progression: Role of the S100A4‐miR155‐SOCS1‐MMP9 axis , 2013, Hepatology.

[41]  Baiyu Chen,et al.  XIST promotes gastric cancer (GC) progression through TGF‐β1 via targeting miR‐185 , 2018, Journal of cellular biochemistry.