Abstract
We aimed to investigate the mechanism and effects of autophagy on cisplatin (DDP)-induced apoptosis in human gastric cancer cell line SGC7901. After SGC7901 cells were treated with DDP and/or chloroquine, cell proliferation was measured using MTT assay; cell apoptosis was determined by flow cytometry; autophagy and apotosis-related proteins expression were detected by Western blot; and quantitative analysis of autophagy after monodansylcadaverine (MDC) staining was performed using fluorescence microscopy. We found after treatment with 5 mg/L DDP for 24 h, the rates of cell apoptosis were (21.07±2.12)%. Autophagy, characterized by an increase in the number of autophagic vesicles and the level of LC3-II protein was observed in cells treated with DDP. After inhibition of autophagy by chloroquine, the rates of cell apoptosis were increased to (30.16±3.54)%, and the level of Caspase-3 and P53 protein were increased, and Bcl-2 protein was decreased. Therefore, autophagy protects human gastric cancer cell line SGC7901 against DDP-induced apoptosis, inhibition of autophagy can promote apoptosis, and combination therapy with DDP and chloroquine may be a promising therapeutic strategy for gastric cancer.
Highlights
Chemotherapy is the main method to cure gastric cancer
We aimed to investigate the mechanism and effects of autophagy on cisplatin (DDP)-induced apoptosis in human gastric cancer cell line SGC7901
After SGC7901 cells were treated with DDP and/or chloroquine, cell proliferation was measured using MTT assay; cell apoptosis was determined by flow cytometry; autophagy and apotosis-related proteins expression were detected by Western blot; and quantitative analysis of autophagy after monodansylcadaverine (MDC) staining was performed using fluorescence microscopy
Summary
Chemotherapy is the main method to cure gastric cancer. As the traditional Chemotherapeutic drug, cisplatin is used throughout all stage of chemotherapy on gastric cancer, such as ECF protocol of adjuvant chemotherapy, DCF protocol of palliative chemotherapy and PF+ Trastuzumab protocol of molecular targeted therapy, all of which are recommended in NCCN Guide as the I type proof (Cunningham et al, 2006; Van Cutsem et al, 2006; Bang et al, 2010). It’s demonstrated in the study that the terminal gastric cancer patients cured by cisplatin protocol gain better therapy effects with longer lifetime (Pasini et al, 2011). It’s reported lately that there is autophagy when cisplatin induces lung cancer cell A549 apoptosis, which will result in cancer cell survive (Ren et al, 2010). There is no domestic and abroad report on whether cisplatin induce the gastric cancer cell autophagy and the influence of autophagy on apoptosis. It’s aimed in the study that the changes of gastric cancer cell SGC7901 are observed before and after autophagy, as well as the influence of autophagy combined inhibitor - cisplatin on autophagy and its mechanism are preliminarily discussed
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