Mobile website
Archive > Volume 27 Issue 8 > 2020,27(8):884-888. DOI:10.3872/j.issn.1007-385X.2020.08.008 Prev Next
Basic Research
Oxaliplatin promotes pyroptosis of colorectal cancer Lovo cells overexpressing GSDME
- Article
- Figures
- Metrics
- Preview PDF
- Reference
- Related
- Cited by
- Materials
Abstract:
Objective: To observe the pyrolysis of colorectal cancer Lovo cells overexpressing Gasdermin E (GSDME) after the treatment with oxaliplatin. Methods: The expression level of GSDME gene in colorectal cancer Lovo cells and normal colorectal epithelial HCOEPIC cells was detected by qPCR. The GSDME-WT (wild-type GSDME) and GSDME-D270A (mutant GSDME) recombinant plasmids were constructed. The plasmids were packaged as lentivirus and then transfected into Lovo cells to construct Lovo cell line with stable and high expression of GSDME. Western blotting was used to detect the expression level of GSDME in cells of WT, D270A and empty vector groups. Different concentrations of oxaliplatin (0, 4, 8, 16, 32, 64 μg/ml) were applied to treat Lovo cells and HCOEPIC cells in WT and D270Agroups, and the morphological changes of the cells were observed under a microscope. Results: The expression of GSDME in HCOEPIC cells was significantly higher than that in Lovo cells (P<0.01). GSDME-WT and GSDME-D270A plasmids with high GSDME expression and the corresponding Lovo cell lines were successfully constructed. Compared with the empty vector group, the expression level of GSDME in Lovo cells of WT and D270A groups were significantly increased (all P<0.05). Observation under the microscope showed that after being treated with 64 μg/ml oxaliplatin for 9 and 12 hours, the volume of Lovo cells and HCOEPIC cells in WT group gradually increased and“blistered”to one side and showed obvious pyrolysis phenomenon. The pyrolysis rate of cells in WT group was significantly higher than that of the control group without oxaliplatin treatment (Lovo cells: [7.405± 1.010]% vs [3.441±0.401]%, P<0.05; HCOEPIC cells: [7.203±1.020]% vs [4.201±0.302]%, P<0.05). Conclusion: Oxaliplatin pro-motes the pyrolysis of colorectal cancer Lovo cells overexpressing GSDME gene.
Keywords:
Project Supported:
Project supported by the Key Grant of Tianjin Health and Family Planning Commission and Tianjin Traditional Chinese Medicine Administration (No.2017057), the National Key Laboratory Open Research Project (No.2018094), and the Science and Technology Support Key Program of Tianjin (No.19YFZCSY00420)
Clc Number:
