DeSUMOylation of protein kinase B1 inhibits cell proliferation and metastasis of hepatocellular carcinoma
10.3760/cma.j.issn.0253-3766.2017.11.003
- VernacularTitle: 蛋白激酶B1去类泛素化修饰抑制肝癌的增殖和转移
- Author:
Feng SHENG
1
;
Yimeng SHEN
2
;
Qiaohao WAN
2
;
Yanxia LI
3
;
Xiaofang MA
3
;
Zhongmin JIANG
4
;
Dianying ZHANG
5
;
Xiaozhi LIU
3
;
Wenhan WU
2
Author Information
1. Department of Traditional Chinese Medicine, the Fifth Central Hospital of Tianjin, Tianjin 300450, China
2. Department of General Surgery, Peking University Frist Hospital, Beijing 100034, China
3. Department of Central Laboratory, the Fifth Central Hospital of Tianjin, Tianjin 300450, China
4. Department of Pathology, the Fifth Central Hospital of Tianjin, Tianjin 300450, China
5. Department of Orthopaedics, Peking University People′s Hospital, Beijing 100044, China
- Publication Type:Journal Article
- Keywords:
Liver neoplasms;
Small ubiquitin-related modifier-1;
Protein kinase B1;
Cell proliferation;
Neoplasm metastasis
- From:
Chinese Journal of Oncology
2017;39(11):814-820
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect of AKT1 deSUMOylation induced by Ubc9 silencing on the proliferation and metastasis of hepatocellular carcinoma (HCC) cells.
Methods:The Ubc9 gene was silenced using RNA interference, and the expression levels of Ubc9, SUMO1 and AKT1 protein were detected by Western blot. Cell proliferation and cell cycle was analyzed by MTT and flow cytometry. Wound healing and transwell assays were used to detect the cell migration ability. Furthermore, the xenograft model was established, and tumor growth curves were drawn. The in situ apoptotic rates was measured using TUNEL Apoptosis Assay. The expression of proliferating cell nuclear antigen (PCNA), matrix metalloproteinase (MMP)-2 and MMP-9 were evaluated by immunohistochemical staining.
Results:Knockdown of Ubc9 gene significantly decreased the protein expression levels of Ubc9, conjugated SUMO1, free SUMO1 and AKT1 in HCC cells (P<0.05 for all). In control, siR-neg and siR-Ubc9 groups, the cell proliferation indexes were 53.19%, 54.25% and 39.17%, respectively. Moreover, cell migration distance and migrating cells per low power field for all these three groups were (59.47±4.66) μm and 89.44±8.36, (56.56±5.37) μm and 93.84±8.79, as well as (34.57±6.61) μm and 41.67±5.39, respectively. In the xenograft model, the weights of subcutaneous tumors for these three groups were (3.78±0.69) g, (3.72±0.72) g and (2.09±0.61) g, respectively. The corresponding apoptotic cell rates were (7.79±2.21)%, (6.45±2.48)% and (33.59±5.44)%, respectively. The expression levels of PCNA, MMP-2 and MMP-9 protein were significantly decreased in siR-Ubc9 group (P<0.05).
Conclusions:Ubc9 silencing in HCC cells induces AKT1 deSUMOylation, and then inhibits the proliferation and metastasis. These results provide a new therapeutic strategy for liver cancer in the future.
