Role Of ZFAS1 Research Articles

Abstract 5429: Therapeutic targeting of oncogenic long non-coding RNAs utilizing antisense oligonucleotides

Abstract Non-coding RNAs (ncRNAs) are emerging as potent regulators of biological process including human diseases such as cancer. Long non-coding RNAs (lncRNAs) are a dynamic and versatile class of ncRNAs for which there is extensive evidence of these transcripts modulating epigenetic programs to affect cellular phenotypes. Our initial studies using a model of glioblastoma (GBM) tumor recurrence has revealed a number of oncogenic lncRNAs associated with therapy resistance and/or tumor recurrence. We have validated the expression of these oncogenic lncRNAs in another cancer, pediatric rhabdomyosarcoma (RMS). There have been few effective therapeutic advances made in past decades for either RMS or GBM. Radiation therapy is commonly employed alongside surgery and standard chemotherapy, especially in the case of unresectable disease, advanced disease, or upon tumor recurrence. Tumors may inherently possess or acquire resistance to radiation therapy and other treatment modalities over the course of disease. Tumor therapy resistance and recurrence are mediated by complex epigenetic programs which lead to increased capacity for DNA damage repair or tolerance of double stranded breaks. LncRNAs may regulate mechanisms of tumor recurrence and therapy resistance across multiple cancer types. The functional roles of ZFAS1 modulating epithelial to mesenchymal transition in colon cancers and SAMMSON which drives aberrant mitochondrial function in melanoma have been described. CASC19/PCAT2, ZFAS1, DUXAP9, and SOX2-OT are overexpressed in multiple cancers with CASC19 having a significant negative association with GBM patient survival. In silico analysis identifies DNA-binding capacities harbored by these lncRNA transcripts, and their predicted binding sites are proximal to several cancer hallmark genes. We have verified the expression of isoforms of these lncRNA transcripts that contain DNA-binding domains through sequencing and qPCR. We hypothesize that these lncRNAs regulate transcriptional networks via recruitment of transcriptional or epigenetic machinery to genomic loci to modulate oncogenic programs and/or tumor suppressor pathways. As potent epigenetic regulators, lncRNAs present an attractive class of therapeutic targets that could complement standard treatments to reduce disease burden and improve patient survival. It is possible that acquired radiation resistance mediated by lncRNAs is conserved across multiple malignancies. We are developing chemically modified, novel antisense oligonucleotides to target these transcripts in GBM and RMS patient-derived xenografts. Using these systems, we hope to reveal novel associations with disease progression and oncogenic mechanisms that could be exploited through the direct targeting of lncRNAs. We hope our work will lead to the development of novel clinical agents to treat patients with RMS, GBM, and potentially other intractable cancers. Citation Format: Christian Tyler Stackhouse, Corinne Linardic, Malaika Kimmons, Samantha Weitzel. Therapeutic targeting of oncogenic long non-coding RNAs utilizing antisense oligonucleotides [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5429.

Oncogenic lncRNA ZNFX1 antisense RNA 1 promotes osteosarcoma cells proliferation and metastasis by stabilizing serine and arginine‑rich splicing factor 3

ABSTRACT Recent studies have demonstrated that lncRNAs play an important role in cancers, particularly osteosarcoma. ZFAS1 is a newly identified and characterized lncRNA linked to a variety of cancers. The role of ZFAS1 in osteosarcoma is mainly unknown. This study discovered that ZFAS1 was upregulated in osteosarcoma patient tissues, which correlates with elevated SRSF3 protein levels. Higher levels of ZFAS1 or SRSF3 were linked to a poor prognosis of osteosarcoma. ZFAS1 knockdown decreased SRSF3 protein levels but had a negligible effect on SRSF3 mRNA expression. Further research indicated that ZFAS1 could bind to the SRSF3 protein directly and prevent degrading. Functional studies revealed that ZFAS1 knockdown inhibited osteosarcoma cell proliferation as measured by the CCK-8 assay, colony formation assay, and Ki-67 immunofluorescence staining. Furthermore, ZFAS1 knockdown reduced the expression of PCNA, CDK1, CDK4, and CDK6, increasing p53 and p16. IT has also been observed that ZFAS1 knockdown inhibited osteosarcoma cell migration and invasion as measured by the wound healing assay and the trans-well assay with or without Matrigel. Furthermore, exogenous SRSF3 expression in ZFAS1-depleted osteosarcoma cells restored SRSF3 expression while simultaneously inhibiting cell proliferation and metastasis. Our findings show that ZFAS1 plays an essential role in osteosarcoma progression by stabilizing the SRSF3 protein. Our study provides novel insight into the role of ZFAS1 in osteosarcoma. ZFAS1 has the potential to be used as a prognostic biomarker as well as a therapeutic target in the treatment of osteosarcoma.

Long non-coding RNA ZFAS1 promotes pancreatic cancer proliferation and metastasis by sponging miR-497-5p to regulate HMGA2 expression

The lncRNA ZFAS1 plays a carcinogenic regulatory role in many human tumours, but it is rarely reported in pancreatic cancer. We identify the role and molecular mechanisms of ZFAS1 in pancreatic cancer. The expression of ZFAS1, miR-497-5p and HMGA2 in pancreatic cancer tissues was detected by qRT-PCR. Pancreatic cancer data in The Cancer Genome Atlas were also included in this study. CCK8, EdU, transwell and scratch wound assays were used to investigate the biological effects of ZFAS1 in pancreatic cancer cells. MS2-RIP, RNA pull-down, RNA-ChIP and luciferase reporter assays were used to clarify the molecular biological mechanisms of ZFAS1 in pancreatic cancer. The role of ZFAS1 in vivo was also confirmed via xenograft experiments. ZFAS1 was overexpressed in pancreatic cancer tissues. ZFAS1 promoted the growth and metastasis of pancreatic cancer cells, and miR-497-5p acted as a tumour suppressor gene in pancreatic cancer by targeting HMGA2. We also demonstrated that ZFAS1 exerts its effects by promoting HMGA2 expression through decoying miR-497-5p. We also found that ZFAS1 promoted the progression of pancreatic cancer in vivo by modulating the miR-497-5p/HMGA2 axis. In conclusion, this study revealed a new role for and the molecular mechanisms of ZFAS1 in pancreatic cancer, identifying ZFAS1 as a novel target for the diagnosis and treatment of pancreatic cancer.

LncRNA ZFAS1 promotes ox-LDL induced EndMT through miR-150-5p/Notch3 signaling axis

EndMT is an active contributor to atherosclerosis pathology, and lncRNAs is widely involved in the occurrence and development of atherosclerosis. The purpose of this study was to investigate the regulatory mechanisms of ZFAS1 in EndMT of atherosclerosis. Here, the ApoE−/− mice were feed with high-fat diet to establish the atherosclerosis model, and HUVECs was stimulated with ox-LDL to induce EndMT. RT-PCR and western blot were used to detect the mRNA and protein expression, respectively. The expression of EndMT markers were detected by immune-fluorescence. The relationships among ZFAS1, miR-150-5p and Notch3 were evaluated by luciferase reporter assay. The role of ZFAS1 in EndMT and its dependence on miR-150-5p/Notch3 axis was further detected by knocking down or over-expressing ZFAS1. We found that ZFAS1 and Notch3 were upregulated while miR-150-5p was downregulated in atherosclerosis mice and ox-LDL-treated HUVECs. The expression of CD31 and vWF were significant decreased, while the α-SMA and vimentin were significant increased in ox-LDL-treated HUVECs, and overexpression of ZFAS1 enhanced the effect of ox-LDL on HUVECs. Further, ZFAS1 functions as a ceRNA to increase Notch3 expression through sponging miR-150-5p, and miR-150-5p mimic or si-Notch3 could reverse LV-ZFAS1-mediated EndMT. In summary, lncRNA ZFAS1 promotes ox-LDL induced HUVECs EndMT through regulating miR-150-5p/Notch3 axis.

LncRNA ZFAS1, as a poor prognostic indicator, promotes cell proliferation and epithelial-mesenchymal transition in endometrial carcinoma.

Background: Long noncoding RNA Zinc finger nuclear transcription factor, X-box binding 1-type containing 1 antisense RNA 1 (ZFAS1) has been reported to be an oncogene in various tumors. However, the role of ZFAS1 in endometrial carcinoma (EC) are not fully determined. Methods & results: Here, we found ZFAS1 expression was significantly upregulated in EC patients, which was significantly associated with International Federation of Gynecology and Obstetrics stage, histological grade, myometrial invasion and poor prognosis. The loss-of-function assays showed that knockdown of ZFAS1 significantly suppressed the proliferation, G1/S transition, migration and invasion in EC cells. Moreover, knockdown of ZFAS1 obviously downregulated the expression of CDK4, Cyclin D1 and N-cadherin, but upregulated E-cadherin expression. Conclusion: Collectively, these results suggest that ZFAS1 might be used as potential therapeutic targets for EC treatment.

Long noncoding RNA expression signature to predict platinum-based chemotherapeutic sensitivity of ovarian cancer patients

Dysregulated long noncoding RNAs (lncRNAs) are potential markers of several tumor prognoses. This study aimed to develop a lncRNA expression signature that can predict chemotherapeutic sensitivity for patients with advanced stage and high-grade serous ovarian cancer (HGS-OvCa) treated with platinum-based chemotherapy. The lncRNA expression profiles of 258 HGS-OvCa patients from The Cancer Genome Atlas were analyzed. Results revealed that an eight-lncRNA signature was significantly associated with chemosensitivity in the multivariate logistic regression model, which can accurately predict the chemosensitivity of patients [Area under curve (AUC) = 0.83]. The association of a chemosensitivity predictor with molecular subtypes indicated the excellent prognosis performance of this marker in differentiated, mesenchymal, and immunoreactive subtypes (AUC > 0.8). The significant correlation between ZFAS1 expression and chemosensitivity was confirmed in 233 HGS-OvCa patients from the Gene Expression Omnibus datasets (GSE9891, GSE63885, and GSE51373). In vitro experiments demonstrated that the ZFAS1 expression was upregulated by cisplatin in A2008, HeyA8, and HeyC2 cell lines. This finding suggested that ZFAS1 may participate in platinum resistance. Therefore, the evaluation of the eight-lncRNA signature may be clinically implicated in the selection of platinum-resistant HGS-OvCa patients. The role of ZFAS1 in platinum resistance should be further investigated.