LncRNA FAM181A-AS1 promotes gliomagenesis by sponging miR-129-5p and upregulating ZRANB2

In this study, we investigated the functional and clinical significance of the long non-coding RNA (lncRNA) FAM181A-AS1 in human gliomas. TCGA, GTEx and CGGA database analyses showed that high FAM181A-AS1 expression correlates with advanced tumor stage and poor survival of glioma patients.
 FAM181A-AS1 expression is higher in glioma cell lines compared to normal human astrocytes (NHA). CCK-8, EdU, and colony formation assays show that FAM181A-AS1 knockdown decreases proliferation and colony formation in glioma cells, whereas, FAM181A-AS1 overexpression reverses these effects. Bioinformatics analysis showed that miR-129-5p is a potential target of FAM181A-AS1.
MiR-129-5p expression negatively correlates with FAM181A-AS1 expression in glioma patients.
Dual-luciferase reporter assays confirmed that miR-129-5p binds directly to FAM181A-AS1 in glioma cells. RNA immunoprecipitation (RIP) assays using anti-Ago2 antibody pulled down FAM181A-AS1 with miR-129-5p.
Bioinformatics analysis identified ZRANB2 as a potential miR-129-5p target gene. Dual luciferase reporter assays confirmed that miR-129-5p binds directly to the 3′-UTR of ZRANBZRANB2 knockdown reduces proliferation and colony formation of FAM181A-AS1 overexpressing glioma cells. These findings show that FAM181A-AS1 promotes gliomagenesis by enhancing ZRANB2 expression by sponging of miR-129-5p.

Arsenite Exposure Displaces Zinc from ZRANB2 Leading to Altered Splicing.

Exposure to arsenic, a class I carcinogen, affects 200 million people globally. Skin is the major target organ but the molecular etiology of arsenic-induced skin carcinogenesis remains unclear. As3+-induced disruption of alternative splicing could be involved, but the mechanism is unknown.
Zinc finger proteins play key roles in alternative splicing. Arsenite (As3+) can displace zinc (Zn2+) from C3H1 and C4 zinc finger motifs (zfms), affecting protein function. ZRANB2, an alternative splicing regulator with two C4 zfms integral to its structure and splicing function was chosen as a candidate for this study.
We hypothesized that As3+ could displace Zn2+ from ZRANB2 altering its structure, expression and splicing function.
As3+/Zn2+ binding and mutual displacement experiments were performed with synthetic apo-peptides corresponding to each ZRANB2 zfm, employing a combination of intrinsic fluorescence, UV spectrophotometry, zinc colorimetric assay and liquid chromatography-tandem mass spectrometry.
ZRANB2 expression in HaCaT cells acutely exposed to As3+ (0 or 5 µM; 0-72 h, or 0-5 M; 6 h) was examined by RT-qPCR and immunoblotting.
ZRANB2-dependent splicing of TRA2B mRNA, a known ZRANB2 target, was monitored by RT-PCR. As3+ bound to, as well as displaced Zn2+ from, each zfm. Also, Zn2+ displaced As3+ from As3+-bound zfms acutely, albeit transiently.
As3+ exposure induced ZRANB2 protein expression between 3-24 h and at all exposures tested, but not ZRANB2 mRNA expression. ZRANB2-directed TRA2B splicing was impaired between 3-24 h post-exposure. Furthermore, ZRANB2 splicing function was also compromised at all As3+ exposures, starting at 100 nm. We conclude that As3+ exposure displaces Zn2+ from ZRANB2 zfms, changing its structure and compromising splicing of its targets, and increases ZRANB2 protein expression as a homeostatic response both at environmental/toxicological exposures and therapeutically relevant doses.

ZRANB2 and SYF2-mediated splicing programs converging on ECT2 are involved in breast cancer cell resistance to doxorubicin.

Besides analyses of specific alternative splicing (AS) variants, little is known about AS regulatory pathways and programs involved in anticancer drug resistance.
Doxorubicin is widely used in breast cancer chemotherapy. Here, we identified 1723 AS events and 41 splicing factors regulated in a breast cancer cell model of acquired resistance to doxorubicin.
An RNAi screen on splicing factors identified the little studied ZRANB2 and SYF2, whose depletion partially reversed doxorubicin resistance. By RNAi and RNA-seq in resistant cells, we found that the AS programs controlled by ZRANB2 and SYF2 were enriched in resistance-associated AS events, and converged on the ECT2 splice variant including exon 5 (ECT2-Ex5+).
Both ZRANB2 and SYF2 were found associated with ECT2 pre-messenger RNA, and ECT2-Ex5+ isoform depletion reduced doxorubicin resistance. Following doxorubicin treatment, resistant cells accumulated in S phase, which partially depended on ZRANB2, SYF2 and the ECT2-Ex5+ isoform.
Finally, doxorubicin combination with an oligonucleotide inhibiting ECT2-Ex5 inclusion reduced doxorubicin-resistant tumor growth in mouse xenografts, and high ECT2-Ex5 inclusion levels were associated with bad prognosis in breast cancer treated with chemotherapy.
Altogether, our data identify AS programs controlled by ZRANB2 and SYF2 and converging on ECT2, that participate to breast cancer cell resistance to doxorubicin.

ZRANB2/SNHG20/FOXK1 Axis regulates Vasculogenic mimicry formation in glioma

Glioma is the most common intracranial neoplasm with vasculogenic mimicry formation as one form of blood supply.
Many RNA-binding proteins and long non-coding RNAs are involved in tumorigenesis of glioma.
The expression of ZRANB2, SNHG20 and FOXK1 in glioma were detected by real-time PCR or western blot.
The function of ZRANB2/SNHG20/FOXK1 axis in glioma-associated with vasculogenic mimicry formation was analyzed.ZRANB2 is up-regulated in glioma tissues and glioma cells. ZRANB2 knockdown inhibits the proliferation, migration, invasion and vasculogenic mimicry formation of glioma cells.
ZRANB2 binds to SNHG20 and increases its stability.
Knockdown of SNHG20 reduces the degradation of FOXK1 mRNA by SMD pathway. FOXK1 inhibits transcription by binding to the promoters of MMP1, MMP9 and VE-Cadherin and inhibits vasculogenic mimicry formation of glioma cells.
ZRANB2/SNHG20/FOXK1 axis plays an important role in regulating vasculogenic mimicry formation of glioma, which might provide new targets of glioma therapy.

Leave a Reply

Your email address will not be published.