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Desumoylating Isopeptidase 2 (DESI2) Inhibits Proliferation and Promotes Apoptosis of Pancreatic Cancer Cells through Regulating PI3K/AKT/mTOR Signaling Pathway

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Pathology & Oncology Research

Abstract

This study aimed to investigate the effects of desumoylating isopeptidase 2 (DESI2) on tumor cell proliferation, apoptosis and invasion of pancreatic cancer, and to assess the signaling pathway involved. Overexpression and silence of DESI2 were designed and the experiments were divided into 5 groups: a normal control group, an interference control group (shRNA-NC); an interference group (sh-DESI2); an overexpression control group (NC), an overexpression group (DESI2). Quantitative real time polymerase chain reaction (qRT-PCR) was used to screen the appropriate interference sequence. The silencing and overexpression of DESI2 were confirmed by qRT-PCR and western blotting. Cell cycling, apoptosis, invasion, and the expression of phosphatidylinositol-3-kinase (PI3K)-protein kinase B (AKT)-mammalian target of rapamycin (mTOR) pathway and caspase 3 were measured. Overexpression and silence of DESI2 were successfully designed in two pancreatic cancer cells, and the interference effect of sh-DESI2–3 showed the best silencing effects. The biological activities of DESI2 were detected in both ASPC-1 and PANCE-1 cells. Our results showed that cell proliferation was significantly increased in the sh-DESI2 group, while decreased in DESI2 group compared with the control group in both cell lines. In ASPC-1 cells, the events in G1 phase decreased and in S phase increased obviously in the sh-DESI2 group, compared with control group. An opposite result was found when DESI2 was overexpressed. In PANCE-1 cells, the events in G2 phase were higher in the sh-DESI2 group, while in the DESI2 group was significantly lower than that in control group. In ASPC-1 and PANCE-1 cells, sh-DESI2 group showed decreased apoptosis, increased cell invasion and increased expression of AKT, p-Akt, PI3K, p-PI3K, p-mTOR and mTOR and decreased caspase 3 expression compared with the control group, while overexpression of DESI2 leaded to increased apoptosis, decreased cell invasion and reduced expression of AKT, p-Akt, PI3K, p-PI3K, p-mTOR and mTOR and increased expression of caspase 3. DESI2 regulates the proliferation and apoptosis of pancreatic cancer cells through PI3K/AKT/mTOR signaling pathway.

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Fig. 1: Confirmation of DESI2 overexpression and interference plasmids using electrophoresis.
Fig. 2: Efficacy of the overexpression and silencing of DESI2.
Fig. 3: Silencing of DESI2 increased, while overexpression of DESI2 reduced cell proliferation of pancreatic cancer cells.
Fig. 4: Silencing of DESI2 increased, while overexpression of DESI2 reduced cell cycling of pancreatic cancer cells.
Fig. 5: Silencing of DESI2 decreased, while overexpression of DESI2 promoted apoptosis of pancreatic cancer cells.
Fig. 6: Silencing of DESI2 increased, while overexpression of DESI2 reduced cell invasion of pancreatic cancer cells.
Fig. 7: Silencing of DESI2 promoted, while overexpression of DESI2 reduced AKT/mTor in APC-1 cells.
Fig. 8: Silencing of DESI2 promoted, while overexpression of DESI2 reduced AKT/mTor in PANCE-1 cells.

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Funding

The present study was supported by the Science and Technology Development Fund Project of Shenzhen (grant no. JCYJ20160428164539088), the Sanming Project of Medicine in Shenzhen (grant no. SZSM201612021) and the Science and Technology Developing Project of Guangdong Province (grant no. 2017B090904010).

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Authors and Affiliations

  1. Department of Hepatopancreatobiliary Surgery, Peking University Shenzhen Hospital, Lian Hua Road 1120, Shenzhen, 518036, China

    Xi Ou, Guang-tao Zhang, Yong Xie, Ji-Kui Liu & Xiao-Ping Liu

  2. The Third Ward of Liver Disease, The Third People’s Hospital of Shenzhen, Shenzhen, 518112, China

    Zhe Xu

  3. Breast Department, Shenzhen Maternity and Child Healthcare Hospital, Shenzhen, 518028, China

    Jing-sen Chen

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  1. Xi Ou
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  2. Guang-tao Zhang
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  4. Jing-sen Chen
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Contributions

XO, GZ, ZX, JC, YX and JL did the experiments and analyzed the data. X Liu YC and XO designed the study and wrote the manuscript.

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Correspondence to Ji-Kui Liu.

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Ou, X., Zhang, Gt., Xu, Z. et al. Desumoylating Isopeptidase 2 (DESI2) Inhibits Proliferation and Promotes Apoptosis of Pancreatic Cancer Cells through Regulating PI3K/AKT/mTOR Signaling Pathway. Pathol. Oncol. Res. 25, 635–646 (2019). https://doi.org/10.1007/s12253-018-0487-4

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