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Expression and mechanisms of long non-coding RNA genes MEG3 and ANRIL in gallbladder cancer

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Tumor Biology

Abstract

The objective of this study was to investigate the expression, proliferation, and apoptosis function of long-chain non-coding RNA maternally expressed gene 3 (MEG3) and antisense non-coding RNA at the INK4 locus (ANRIL) in gallbladder cancer (GBC) tissues. GBC tissues and adjacent normal samples were collected from 84 patients from January 2008 to June 2010. Empty vector, pcDNA-MEG3, and pcDNA-ANRIL vectors were transfected into GBC-SD and QBC939 cells. An MTT assay, real-time quantitative polymerase chain reaction (RT-qPCR), flow cytometry, Western blotting, and immunohistochemistry were applied. The effects of MEG3 and ANRIL were also verified in mice. Compared with normal tissues, the expression of MEG3 was significantly lower in GBC tissues, whereas the expression of ANRIL was significantly higher (both P < 0.05). The overexpression of MEG3 and underexpression of ANRIL were significantly associated with GBC prognosis (both P < 0.05). The expressions of MEG3 and ANRIL were higher in pcDNA-MEG3 and pcDNA-ANRIL-transfected cells than in empty vector-transfected cells in vitro (both P < 0.05). Most of the pcDNA-MEG3-transfected cells were in the G0-G1 phase, which showed reduced cell activity and clone counts and increased p53 and decreased cyclin D1, whereas the pcDNA-ANRIL-transfected cells were mostly in the S phase and showed contrasting behavior. Mice injected with pcDNA-MEG3-transfected cells had smaller and lighter tumors, decreased ki-67 levels, and increased caspase 3 levels, whereas those injected with pcDNA-ANRIL showed contrasting results (all P < 0.05). MEG3 can inhibit the proliferation of GBC cells and promote apoptosis, whereas ANRIL can improve the proliferation of gallbladder cells and inhibit apoptosis. Collectively, our results suggest that therapeutic strategies directed toward upregulating MEG3 and downregulating ANRIL may be clinically relevant for the inhibition of GBC deterioration.

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Acknowledgments

This project was supported by the National Natural Science Foundation of China (No: 81302075). We would like to acknowledge the reviewers for their helpful comments on this paper.

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Authors and Affiliations

  1. Department of General Surgery, Xiangya No. 2 Hospital, Central South University, Renmin Road No 139, Furong Area, Changsha, 410000, People’s Republic of China

    Bo Liu & Wei-Dong Chen

  2. Department of Oncology, No. 1 People’s Hospital of Yueyang City, Yueyang, 414000, People’s Republic of China

    Er-Dong Shen

  3. Department of General Surgery, Xiangya Hospital, Central South University, Changsha, 410000, People’s Republic of China

    Ming-Mei Liao

  4. Department of Pathology, Xiangya Hospital, Central South University, Changsha, 410000, People’s Republic of China

    Yong-Bin Hu

  5. Department of Physiatry, Xiangya Hospital, Central South University, Changsha, 410000, People’s Republic of China

    Kai Wu

  6. Department of Vascular Surgery, Xiangya Hospital, Central South University, Changsha, 410000, People’s Republic of China

    Pu Yang

  7. Department of Colorectal and Anal Surgery, Xiangya Hospital, Central South University, Changsha, 410000, People’s Republic of China

    Lin Zhou

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  1. Bo Liu
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Correspondence to Wei-Dong Chen.

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All in vivo studies were carried out in strict accordance with the recommendations of the Animal Experiment Association.

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The study was approved by the Institute Review Board of Xiangya No. 2 Hospital of Central South University and fulfilled the ethical principles outlined in the Declaration of Helsinki. All patients and their family members provided written informed consent.

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Liu, B., Shen, ED., Liao, MM. et al. Expression and mechanisms of long non-coding RNA genes MEG3 and ANRIL in gallbladder cancer. Tumor Biol. 37, 9875–9886 (2016). https://doi.org/10.1007/s13277-016-4863-y

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  • DOI: https://doi.org/10.1007/s13277-016-4863-y

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