Yang Sun1, Jie Zhang, Zhi-lan Peng. 1. Department of Obstetrics and Gynecology, West China Second Hospital, West China Center of Medical Sciences, Sichuan University, Chengdu, Sichuan, China.
Abstract
BACKGROUND: To investigate the role of Beclin1 in the relationship between autophagy and apoptosis after carboplatin treatment. METHODS: After overexpression or partial silencing of Beclin1 in cervical cancer SiHa cells, the transfected group and the control group were treated with carboplatin for 48 hours. The expression of Beclin1 and LC3 was measured using a western blot. The ultrastructural analysis was conducted with an electron microscope. Fluorescent changes in autophagic cells were observed using reverse fluorescent microscopy. The percentage of apoptotic cells and autophagic cells and cell proliferation were assessed by flow cytometry and MTT assay. RESULTS: Expression of LC3 and Beclin1 protein was up-regulated in overexpressed transfectants of SiHa cells. After treatment with carboplatin for 48 hours, the flow cytometric analysis indicated that the Beclin1 transfected group showed a greater increase in apoptosis and autophagic cells than did the nontransfected group. MTT assay showed that carboplatin inhibited proliferation of SiHa cells in Beclin1-overexpressed transfectant cells more than in nontransfectant cells. CONCLUSIONS: These results suggest that autophagy and apoptosis had differential contributions to carboplatin-induced death of cervical cancer SiHa cells. Overexpression of Beclin1 in SiHa cells may enhance apoptosis signaling induced by carboplatin.
BACKGROUND: To investigate the role of Beclin1 in the relationship between autophagy and apoptosis after carboplatin treatment. METHODS: After overexpression or partial silencing of Beclin1 in cervical cancer SiHa cells, the transfected group and the control group were treated with carboplatin for 48 hours. The expression of Beclin1 and LC3 was measured using a western blot. The ultrastructural analysis was conducted with an electron microscope. Fluorescent changes in autophagic cells were observed using reverse fluorescent microscopy. The percentage of apoptotic cells and autophagic cells and cell proliferation were assessed by flow cytometry and MTT assay. RESULTS: Expression of LC3 and Beclin1 protein was up-regulated in overexpressed transfectants of SiHa cells. After treatment with carboplatin for 48 hours, the flow cytometric analysis indicated that the Beclin1 transfected group showed a greater increase in apoptosis and autophagic cells than did the nontransfected group. MTT assay showed that carboplatin inhibited proliferation of SiHa cells in Beclin1-overexpressed transfectant cells more than in nontransfectant cells. CONCLUSIONS: These results suggest that autophagy and apoptosis had differential contributions to carboplatin-induced death of cervical cancer SiHa cells. Overexpression of Beclin1 in SiHa cells may enhance apoptosis signaling induced by carboplatin.
Authors: Mitzie-Ann Davis; Joe R Delaney; Chandni B Patel; Ryan Storgard; Dwayne G Stupack Journal: Drug Des Devel Ther Date: 2016-06-02 Impact factor: 4.162