Ou Xi, Zhang Guang-Tao, Xu Zhe, Chen Jing-Sen, Xie Yong, Liu Ji-Kui, Liu Xiao-Ping
Department of Hepatopancreatobiliary Surgery, Peking University Shenzhen Hospital, Lian Hua Road 1120, Shenzhen, 518036, China.
The Third Ward of Liver Disease, The Third People's Hospital of Shenzhen, Shenzhen, 518112, China.
Pathol Oncol Res. 2019 Apr;25(2):635-646. doi: 10.1007/s12253-018-0487-4. Epub 2018 Nov 8.
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.
本研究旨在探讨去SUMO化异肽酶2（DESI2）对胰腺癌细胞增殖、凋亡及侵袭的影响，并评估其涉及的信号通路。设计了DESI2的过表达和沉默实验，实验分为5组：正常对照组、干扰对照组（shRNA-NC）；干扰组（sh-DESI2）；过表达对照组（NC）、过表达组（DESI2）。采用定量实时聚合酶链反应（qRT-PCR）筛选合适的干扰序列。通过qRT-PCR和蛋白质免疫印迹法证实DESI2的沉默和过表达。检测细胞周期、凋亡、侵袭以及磷脂酰肌醇-3-激酶（PI3K）-蛋白激酶B（AKT）-雷帕霉素靶蛋白（mTOR）信号通路和半胱天冬酶3的表达。在两种胰腺癌细胞中成功设计了DESI2的过表达和沉默，sh-DESI2-3的干扰效果显示出最佳的沉默效果。在ASPC-1和PANCE-1细胞中均检测到DESI2的生物学活性。我们的结果表明，与对照组相比，sh-DESI2组细胞增殖显著增加，而DESI2组细胞增殖减少。在ASPC-1细胞中，与对照组相比，sh-DESI2组G1期事件减少，S期事件明显增加。当DESI2过表达时发现相反的结果。在PANCE-1细胞中，sh-DESI2组G2期事件较高，而DESI2组明显低于对照组。在ASPC-1和PANCE-1细胞中，与对照组相比，sh-DESI2组凋亡减少、细胞侵袭增加、AKT、p-Akt、PI3K、p-PI3K、p-mTOR和mTOR表达增加，半胱天冬酶3表达减少，而DESI2过表达导致凋亡增加、细胞侵袭减少、AKT、p-Akt、PI3K、p-PI3K、p-mTOR和mTOR表达降低，半胱天冬酶3表达增加。DESI2通过PI3K/AKT/mTOR信号通路调节胰腺癌细胞的增殖和凋亡。
