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LINC00998 functions as a novel tumor suppressor in acute myeloid leukemia via regulating the ZFP36 ring finger protein/mammalian target of rapamycin complex 2 axis

Fang, Ximin, Pan, Xiazhen, Mai, Huirong, Yuan, Xiuli, Liu, Sixi, Wen, Feiqiu·2021·Bioengineered
DOI 10.1080/21655979.2021.1996506PMC PMC8810020
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Figure 1. LINC00998 is lowly expressed in human AML. (a) GEPIA database showing the expression of LINC00998 in AML. (b, c) qRT-PCR analysis of LINC00998 in PBMCs from 80 AML patients and 59 healthy controls, followed by ROC curve analysis. (d, e) qRT-PCR analysis of LINC00998 in PBMCs from AML patients with or without relapse, followed by ROC curve analysis. (f) The survival curve of AML patients with low or high LINC00998 level based on median LINC00998 expression (Hazard Ratio = 3.161, 95% confidence interval = 1.565 ~ 6.386). (g) qRT-PCR analysis of LINC00998 level in nuclear and cytoplasmic fragments of U937 and THP-1 cells. ***P < 0.001
Figure 1. LINC00998 is lowly expressed in human AML. (a) GEPIA database showing the expression of LINC00998 in AML. (b, c) qRT-PCR analysis of LINC00998 in PBMCs from 80 AML patients and 59 healthy controls, followed by ROC curve analysis. (d, e) qRT-PCR analysis of LINC00998 in PBMCs from AML patients with or without relapse, followed by ROC curve analysis. (f) The survival curve of AML patients with low or high LINC00998 level based on median LINC00998 expression (Hazard Ratio = 3.161, 95% confidence interval = 1.565 ~ 6.386). (g) qRT-PCR analysis of LINC00998 level in nuclear and cytoplasmic fragments of U937 and THP-1 cells. ***P < 0.001CC BY · PubMed Central
Figure 2. LINC00998 inhibits AML cell malignant behaviors. (a) qRT-PCR verifying the overexpression of LINC00998 in U937 cells. (b–d) CCK-8, colony formation and EdU assays testing cell viability, colony and DNA synthesis rate of U937 cells after LINC00998 overexpression. (e–h) The effect of LINC00998 overexpression on cell viability, colony and DNA synthesis rate. (i) Flow cytometry analyzing cell apoptosis after LINC00998 overexpression. (j) The indicated protein levels in LINC00998-overexpressing AML cell lines. Scale bar = 50 μm, **P < 0.01
Figure 2. LINC00998 inhibits AML cell malignant behaviors. (a) qRT-PCR verifying the overexpression of LINC00998 in U937 cells. (b–d) CCK-8, colony formation and EdU assays testing cell viability, colony and DNA synthesis rate of U937 cells after LINC00998 overexpression. (e–h) The effect of LINC00998 overexpression on cell viability, colony and DNA synthesis rate. (i) Flow cytometry analyzing cell apoptosis after LINC00998 overexpression. (j) The indicated protein levels in LINC00998-overexpressing AML cell lines. Scale bar = 50 μm, **P < 0.01CC BY · PubMed Central
Figure 3. LINC00998 inhibits mTORC2 level. (a) The correlation between LINC00998 and mTORC2 mRNA in TCGA AML database. (b, c). The mRNA and protein levels of mTORC2 after LINC00998 overexpression. (d) qRT-PCR analysis of mTORC2 mRNA levels in PBMCs from 80 AML patients and 59 healthy controls. (e) The correlation between LINC00998 and mTORC2 mRNA in AML. F-H. Cell viability, colony and DNA synthesis in LINC00998-overexpressed U937 and THP-1 cells with mTORC2 overexpression. **P < 0.01, ***P < 0.001
Figure 3. LINC00998 inhibits mTORC2 level. (a) The correlation between LINC00998 and mTORC2 mRNA in TCGA AML database. (b, c). The mRNA and protein levels of mTORC2 after LINC00998 overexpression. (d) qRT-PCR analysis of mTORC2 mRNA levels in PBMCs from 80 AML patients and 59 healthy controls. (e) The correlation between LINC00998 and mTORC2 mRNA in AML. F-H. Cell viability, colony and DNA synthesis in LINC00998-overexpressed U937 and THP-1 cells with mTORC2 overexpression. **P < 0.01, ***P < 0.001CC BY · PubMed Central
Figure 4. LINC00998 enhances ZFP36-mediated mTORC2 mRNA decay. (a) The promoter activity of mTORC2 affected by LINC00998. (b) qRT-PCR analysis of mTORC2 mRNA level at the indicated time in LINC00998-overexpressed AML cells treated with Actinomycin D. (c) qRT-PCR testing the effect of ZFP36 silencing on mTORC2 expression in LINC00998-overexpressed AML cells. (d) The enrichment of mTORC2 3`-UTR by ZFP36 antibody. (e) RNA pull-down assay using LINC00998 probe, followed by western blot analysis of ZFP36 level. (f) The enrichment of LINC00998 by ZFP36 antibody. *P < 0.05, **P < 0.01
Figure 4. LINC00998 enhances ZFP36-mediated mTORC2 mRNA decay. (a) The promoter activity of mTORC2 affected by LINC00998. (b) qRT-PCR analysis of mTORC2 mRNA level at the indicated time in LINC00998-overexpressed AML cells treated with Actinomycin D. (c) qRT-PCR testing the effect of ZFP36 silencing on mTORC2 expression in LINC00998-overexpressed AML cells. (d) The enrichment of mTORC2 3`-UTR by ZFP36 antibody. (e) RNA pull-down assay using LINC00998 probe, followed by western blot analysis of ZFP36 level. (f) The enrichment of LINC00998 by ZFP36 antibody. *P < 0.05, **P < 0.01CC BY · PubMed Central
Figure 5. LINC00998 inhibits tumor growth in vivo. (a–c). Tumor volume, weight and image in control and LINC00998-overexpressing groups. (d) qRT-PCR analysis of LINC00998 and mTORC2 levels in the indicated two groups. (e) Western blot analyzing mTORC2 and Ki-67 levels in the indicated two groups. (f) TUNEL staining testing cell apoptosis in the indicated two groups. Staining of nucleus with DAPI. Scale bar = 50 μm, **P < 0.01, ***P < 0.001
Figure 5. LINC00998 inhibits tumor growth in vivo. (a–c). Tumor volume, weight and image in control and LINC00998-overexpressing groups. (d) qRT-PCR analysis of LINC00998 and mTORC2 levels in the indicated two groups. (e) Western blot analyzing mTORC2 and Ki-67 levels in the indicated two groups. (f) TUNEL staining testing cell apoptosis in the indicated two groups. Staining of nucleus with DAPI. Scale bar = 50 μm, **P < 0.01, ***P < 0.001CC BY · PubMed Central

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Metadata

Authors
Fang, X., Pan, X., Mai, H., Yuan, X., Liu, S., & Wen, F.
Affiliations
  • The First Affiliated Hospital of Jinan University
  • The University of Hong Kong-Shenzhen Hospital
  • Shenzhen Children’s Hospital
ORCID
Wen, Feiqiu
Venue
Bioengineered
Publisher
Informa UK Limited
Published
2021-12-02
Volume
12
Issue
2
Pages
10363-10372
ISSN
  • 2165-5979
  • 2165-5987
DOI
10.1080/21655979.2021.1996506
PMC
PMC8810020
Cited by
10
References
0
Language
en
Licence
doi.org/10.1080/policies-text-data-mining (tdm)
Funders
Shenzhen Science and Technology Project, Guangdong Province, China (JCYJ20160429180540076)

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CoChat. (2026). Citation check: LINC00998 functions as a novel tumor suppressor in acute myeloid leukemia via regulating the ZFP36 ring finger protein/mammalian target of rapamycin complex 2 axis. https://app.cochat.ai/papers/10.1080/21655979.2021.1996506
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Fang, X., Pan, X., Mai, H., Yuan, X., Liu, S., & Wen, F. (2021). LINC00998 functions as a novel tumor suppressor in acute myeloid leukemia via regulating the ZFP36 ring finger protein/mammalian target of rapamycin complex 2 axis. Bioengineered. https://doi.org/10.1080/21655979.2021.1996506