IMB5036 inhibits human pancreatic cancer growth primarily through activating necroptosis
Qi Zhao
Department of Pharmacology, Shanxi Medical University, Key Laboratory of Cellular Physiology (Shanxi Medical University), Ministry of Education, Taiyuan, China
Department of Oncology, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China
Search for more papers by this authorYanbo Zheng
Department of Oncology, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China
Search for more papers by this authorXing Lv
Department of Biochemistry and Molecular Biology, Shanxi Medical University, Key Laboratory of Cellular Physiology (Shanxi Medical University), Ministry of Education, Taiyuan, China
Search for more papers by this authorCorresponding Author
Jianhua Gong
Department of Oncology, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China
Correspondence
Jianhua Gong, Department of Oncology, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Tiantan Xili, 100050, Beijing, China.
Email: [email protected]
Lijun Yang, Department of Pharmacology, Shanxi Medical University, Key Laboratory of Cellular Physiology (Shanxi Medical University), Ministry of Education. Taiyuan, 030001, Shanxi, China.
Email: [email protected]
Search for more papers by this authorCorresponding Author
Lijun Yang
Department of Pharmacology, Shanxi Medical University, Key Laboratory of Cellular Physiology (Shanxi Medical University), Ministry of Education, Taiyuan, China
Correspondence
Jianhua Gong, Department of Oncology, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Tiantan Xili, 100050, Beijing, China.
Email: [email protected]
Lijun Yang, Department of Pharmacology, Shanxi Medical University, Key Laboratory of Cellular Physiology (Shanxi Medical University), Ministry of Education. Taiyuan, 030001, Shanxi, China.
Email: [email protected]
Search for more papers by this authorQi Zhao
Department of Pharmacology, Shanxi Medical University, Key Laboratory of Cellular Physiology (Shanxi Medical University), Ministry of Education, Taiyuan, China
Department of Oncology, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China
Search for more papers by this authorYanbo Zheng
Department of Oncology, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China
Search for more papers by this authorXing Lv
Department of Biochemistry and Molecular Biology, Shanxi Medical University, Key Laboratory of Cellular Physiology (Shanxi Medical University), Ministry of Education, Taiyuan, China
Search for more papers by this authorCorresponding Author
Jianhua Gong
Department of Oncology, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China
Correspondence
Jianhua Gong, Department of Oncology, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Tiantan Xili, 100050, Beijing, China.
Email: [email protected]
Lijun Yang, Department of Pharmacology, Shanxi Medical University, Key Laboratory of Cellular Physiology (Shanxi Medical University), Ministry of Education. Taiyuan, 030001, Shanxi, China.
Email: [email protected]
Search for more papers by this authorCorresponding Author
Lijun Yang
Department of Pharmacology, Shanxi Medical University, Key Laboratory of Cellular Physiology (Shanxi Medical University), Ministry of Education, Taiyuan, China
Correspondence
Jianhua Gong, Department of Oncology, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Tiantan Xili, 100050, Beijing, China.
Email: [email protected]
Lijun Yang, Department of Pharmacology, Shanxi Medical University, Key Laboratory of Cellular Physiology (Shanxi Medical University), Ministry of Education. Taiyuan, 030001, Shanxi, China.
Email: [email protected]
Search for more papers by this authorQi Zhao and Yanbo Zheng equally contributed to this work.
Funding information: National Natural Science Foundation of China, Grant/Award Number: 81972325; Fundamental Research Funds for the Central Universities, Grant/Award Number: 2016ZX350056; CAMS Innovation Fund for Medical Sciences, Grant/Award Number: 2021-1-I2M-030
Abstract
IMB5036 is a novel pyridazinone compound with potent cytotoxicity. In this study, we reported its antitumour activity against pancreatic cancer and the underlying mechanism. We found that IMB5036 induced rapid cell swelling and increased membrane permeability in pancreatic cancer cells. IMB5036 increased the ratio of PI+ cells, which could be rescued by necroptosis inhibitor. Furthermore, MLKL inhibitor NSA attenuated the killing effect of IMB5036 on pancreatic cancer cells. IMB5036 stimulated translocation of MLKL and p-MLKL from cytoplasm to cell membrane. IMB5036 upregulated the level of p-RIPK1, p-RIPK3, and p-MLKL. At the same time, IMB5036 also partially activated apoptosis and pyroptosis. IMB5036 inhibited tumour growth in pancreatic xenograft. IMB5036 induced larger necrosis area, increased p-MLKL level, and inhibited Ki67 expression in tumour mass. The study indicates that IMB5036 inhibits human pancreatic cancer growth primarily activating necroptosis.
CONFLICT OF INTEREST
The authors declare no conflict of interest.
Open Research
DATA AVAILABILITY STATEMENT
The datasets used in the current study are available from the corresponding author on reasonable request.
Supporting Information
| Filename | Description |
|---|---|
| bcpt13694-sup-0001-Figure_S1-S2.docxWord 2007 document , 1.1 MB |
FIGURE S1. IMB5036 increased membrane permeability. IMB5036 treatment significantly induced cells swelling in MIAPaCa-2 and Capan-2 cells. Cells were treated with IMB5036 for 4 h and observed by bright field microscopy (Scale bar = 40 μm). Data were based on at least three independent experiments. |
| bcpt13694-sup-0001-Figure_S1-S2.docxWord 2007 document , 1.1 MB |
FIGURE S2. The effect of IMB5036 on cell apoptosis and cell cycle. (A) Flow cytometry analysis of apoptosis in MIA PaCa-2 cells treated with IMB5036. Cells were treated with IMB5036 for 24 h, following Annexin V/PI staining. (B) Expression level of pro-caspase-3 and PARP assessed by Western blotting for the MIAPaCa-2 cells treated with IMB5036 for 24 h. (C-D) Expression level of pro-caspase-3 and PARP, GSDME, GSDMD assessed by Western blotting for the MIAPaCa-2 cells treated with IMB5036 (5 μM) for 2 h,6 h. (E) The distribution of cell cycle was detected by flow cytometry after PI staining in MIAPaCa-2 cells. Quantitative analysis of the number of cells in different phases of the cell cycle treated with IMB5036. Data were based on at least three independent experiments. |
| bcpt13694-sup-0002-Video_S1.avivideo/avi, 17.9 MB |
Video S1. IMB5036 rapidly induces membrane blebbing and permeabilization of MIAPaCa-2 cells. Cells were imaged over a period of 40 min, with IMB5036 (10 μM) being added to cells at 30 min. |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
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