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LINC00313 facilitates osteosarcoma carcinogenesis and metastasis through enhancing EZH2 mRNA stability and EZH2-mediated silence of PTEN expression

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Abstract

Background

Osteosarcoma is one of the five leading causes of cancer death among all pediatric malignancies. Recent advances in non-coding RNAs suggested that many long noncoding RNAs (lncRNAs) are dysregulated in cancer tissues and play important roles in carcinogenesis. We aimed to further explore the mechanisms of Long Intergenic Non-Protein Coding RNA 313 (LINC00313)-promoted malignant phenotypes of osteosarcoma.

Methods

The mRNA expressions were determined by quantitative real-time PCR. Protein levels were detected using Western blotting or immunohistochemistry staining. Protein binding to genomic DNA and RNA were measured using chromatin and RNA immunoprecipitation assay, respectively. CCK-8 and EdU incorporation assay were adopted to detect cell proliferation. Transwell assay was employed to assess the capacity of cell migration and invasion. The roles of LINC00313 and its target genes in tumorigenesis and metastasis of osteosarcoma were evaluated using subcutaneous xenograft models and tail vein inoculation models.

Results

LINC00313 was elevated in osteosarcoma tissues compared with adjacent normal tissues. Higher LINC00313 was associated with advanced grades of osteosarcoma. LINC00313 promoted cell proliferation, migration, invasion in vitro and tumor growth as well as metastasis in vivo through inhibiting PTEN expression to promote AKT phosphorylation. Mechanistically, LINC00313 favored the interaction between FUS and EZH2, leading to the prolonged half-life of EZH2 mRNA, thereby in turn up-regulating EZH2 proteins and increasing EZH2-mediated epigenetic silence of PTEN.

Conclusion

LINC00313 exerted oncogene-like actions through increasing EZH2 mRNA stability, leading to PTEN deficiency in osteosarcoma.

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Availability of data and material

The datasets used or analyzed during the current study are available from the corresponding author on reasonable request.

Code availability

Not applicable.

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Acknowledgements

We thank Hunan UCbio Co., Ltd. for providing technical support for animal experiments.

Funding

This work was supported by Natural Science Foundation of Zhejiang Province (LY19H160047).

Author information

Authors and Affiliations

  1. Department of Orthopedics, First Affiliated Hospital, College of Medicine, Zhejiang University, No. 79, Qingchun Road, Hangzhou, 310003, Zhejiang Province, China

    Chun-Yang Xing & Yu-Zhu Zhang

  2. Department of Radiotherapy, The People’s Hospital of Changxing County, Huzhou, 313199, Zhejiang Province, China

    Wei Hu

  3. Department of Emergency, Qilu Hospital, Shandong University, Jinan, 250063, Shandong Province, China

    Li-Yuan Zhao

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  1. Chun-Yang Xing
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  2. Yu-Zhu Zhang
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  3. Wei Hu
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Corresponding author

Correspondence to Chun-Yang Xing.

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Conflict of interest

The authors declare no conflicts of interest.

Ethics approval and consent to participate

The specimen collection and processing procedures were approved by the research ethics committee of The First Affiliated Hospital, Zhejiang University. All patients were informed of the study and signed the written consent. All the procedures of animal experiments were approved by the research ethics committee of The First Affiliated Hospital, Zhejiang University.

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Supplementary Information

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Supplementary Figure 1. A

Time-series study of LINC00313 post transfection of sh-LINC00313 plasmid in HOS and 143B cells. B Time-series study of LINC00313 post transfection of pcDNA-LINC00313 plasmid in HOS and 143B cells. C Time-series qRT-PCR of PTEN detection post transfection of siPTEN in HOS and 143B cells. D Time-series qRT-PCR of PTEN detection post transfection of pcDNA-PTEN in HOS and 143B cells. E Time-series Western blotting of PTEN detection post transfection of siPTEN in HOS and 143B cells. F Time-series Western blotting of PTEN detection post transfection of pcDNA-PTEN in HOS and 143B cells. G Time-series qRT-PCR of FUS detection post transfection of sh-FUS in HOS and 143B cells. H Time-series qRT-PCR of FUS detection post transfection of pcDNA-FUS in HOS and 143B cells. I Time-series Western blotting of FUS detection post transfection of sh-FUS in HOS and 143B cells. J Time-series Western blotting of FUS detection post transfection of pcDNA-FUS in HOS and 143B cells. K Comparison of LINC00313 knockdown efficiencies between sh-LINC00313 and sh-LINC00313+siPTEN. Comparison of LINC00313 levels between pcDNA-LINC00313 and pcDNA-LINC00313+pcDNA-PTEN in HOS and 143B cells. L Comparison of LINC00313 knockdown efficiencies between sh-LINC00313 and sh-LINC00313+pcDNA-FUS. Comparison of LINC00313 levels between pcDNA-LINC00313 and pcDNA-LINC00313+sh-FUS in HOS and 143B cells. M Comparison of LINC00313 levels between pcDNA-LINC00313 single transfection and co-transfection with other plasmids in HOS cells. N=3. Error bars represented mean ± Standard deviation (SD). *p<0.05, **p<0.01, ***p<0.001 (TIF 1748 KB)

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Xing, CY., Zhang, YZ., Hu, W. et al. LINC00313 facilitates osteosarcoma carcinogenesis and metastasis through enhancing EZH2 mRNA stability and EZH2-mediated silence of PTEN expression. Cell. Mol. Life Sci. 79, 382 (2022). https://doi.org/10.1007/s00018-022-04376-1

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