Study on abnormal activation and targeted therapy of SUMO pathway in osteosarcoma
10.3760/cma.j.cn121113-20200415-00258
- VernacularTitle:小泛素样修饰蛋白通路在骨肉瘤中异常激活与靶向治疗的研究
- Author:
Kang YU
1
;
Xiuxin HAN
;
Guowen WANG
;
Yulin MA
;
Chao ZHANG
;
Jin ZHANG
;
Chao ZHANG
;
Lili LI
Author Information
1. 天津医科大学肿瘤医院骨与软组织肿瘤科，国家肿瘤临床医学研究中心，天津市 肿瘤防治重点实验室，天津市恶性肿瘤临床医学研究中心 300060
- From:
Chinese Journal of Orthopaedics
2020;40(13):864-872
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To observe the expression characteristics of small ubiquitin-like modified protein (SUMO) pathway members in osteosarcomaby using molecular biology methods, and provide theoretical basis for targeted therapy based on protein SUMO modification.Methods:Eighteen fresh osteosarcoma tissue samples surgically resected at Tianjin Cancer Hospital from January 2017 to June 2019 were collected. Western blot and immunohistochemical methods were used to detect the protein expressions of SUMO1, SAE1, Ubc9, and SENP1, which are core members of SUMO pathway, in the cancerous and adjacent tissue. Taking osteosarcoma cell line 143B as the research object, three targeted treatment regimens were designed and grouped as follows: control group, nonsense group, siR-SUMO1 group, siR-Ubc9 group, and SENP1 group. Western blot was used to verify the efficiency of gene transfection, the positive expression rate of EdU was detected by cell proliferation detection kit, the ability of cell migration and invasion was measured by scratch test and Transwell invasion test, and the apoptosis rate was detected by flow cytometry. Bone marrow mesenchymal stem cells (BMSCs) were used to evaluate the side effects of three treatment regimens.Results:Results from Western blot and immunohistochemistry showed that the protein expression levels of SUMO1, SAE1, andUbc9 in osteosarcoma tissues were significantly higher than those in adjacent tissues ( P<0.05), but SENP1 was significantly lower than in adjacent tissues. The experimental results based on 143B osteosarcoma cells showed that the siR-SUMO1 group, siR-Ubc9 group, and SENP1 group can significantly inhibit the activation of SUMO pathway in osteosarcoma cells, showing lower tumor cell proliferation ( P<0.05), slower cell migration and invasion capacity ( P<0.05), and higher apoptotic rate ( P<0.05), than the control group and nonsense group; however, experimental results based on BMSCs showed that the siR-SUMO1 group and siR-Ubc9 group can significantly inhibit the proliferation of BMSCs, induce apoptosis, and show great therapeutic side effects, but the SENP1 group has almost no effect on the proliferation and apoptosis of BMSCs ( P>0.05). Conclusion:The SUMO pathway is abnormally activated in osteosarcoma. Exogenous supplementation or endogenous activation of SENP1 is one of the alternatives for targeted osteosarcoma treatment.
