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BI2536 enhances the DNA damage effect of DDP in ESCC cells. (a), KYSE510 cells treated with BI2536 (20 nM), DDP (10 μM), or the combination of the two drugs for 24 h were fixed and co-labelled with anti-γH2AX and anti-RAD51 antibodies and DAPI. The γ-H2AX and RAD51 foci were analysed by immunofluorescence microscopy. (b), KYSE510 cells were treated as described in (a) and analysed via comet assay. At the bottom left is the enlarged image. (c), The percentage of cells positive for γ-H2AX and RAD51, which had N5 foci, was calculated. The results are displayed as the mean ± SEM from a minimum of 200 cells per condition for three independent experiments. *, P b .05. (d), Olive moment values were normalized to those of KYSE150 cells receiving no treatment. The results are shown as the mean ± SEM from one of three independent experiments. A minimum of 50 cells per experiment were analysed. ***, P b .05. (e), The expression of the indicated markers reflecting DNA damage and repair was examined using Western blotting. Lysates from KYSE150 (left) and KYSE510 (right) cells were probed with antibodies after receiving the same treatment described in (a).
Source publication
Background
Targeting PLK1 has recently been proven as a viable therapeutic strategy against oesophageal squamous cell carcinom (ESCC). Therefore, this study aimed to explore whether the PLK1 inhibitor BI2536 is able to sensitize ESCC cells to cisplatin (DDP) and determine the underlying mechanisms.
Methods
Viability, clonogenicity, cell cycle dist...
Contexts in source publication
Context 1
... and DDP, we employed immunofluorescence to analyse the coexpression of γ-H2AX and RAD51, which are markers of DNA doublestrand breaks (DSBs) and DNA damage repair (DDR), respectively. We observed a very large fraction of cells that were positive for γ-H2AX staining but negative for RAD51 staining after 24 h of treatment with BI2536 and DDP ( Fig. 4A and C, Supplementary file: Fig. S4A and C). These results suggested that BI2536 increased the DNA damage effect of DDP and decreased the DDR ability of ESCC cells. Single-cell gel electrophoresis or comet assays were performed to assess the DNA damage induced by BI2536 and DDP. The results showed that both monotherapy and combination ...
Context 2
... to analyse the coexpression of γ-H2AX and RAD51, which are markers of DNA doublestrand breaks (DSBs) and DNA damage repair (DDR), respectively. We observed a very large fraction of cells that were positive for γ-H2AX staining but negative for RAD51 staining after 24 h of treatment with BI2536 and DDP ( Fig. 4A and C, Supplementary file: Fig. S4A and C). These results suggested that BI2536 increased the DNA damage effect of DDP and decreased the DDR ability of ESCC cells. Single-cell gel electrophoresis or comet assays were performed to assess the DNA damage induced by BI2536 and DDP. The results showed that both monotherapy and combination therapy could cause DNA strand breaks, as ...
Context 3
... of DDP and decreased the DDR ability of ESCC cells. Single-cell gel electrophoresis or comet assays were performed to assess the DNA damage induced by BI2536 and DDP. The results showed that both monotherapy and combination therapy could cause DNA strand breaks, as indicated by the higher frequency of Olive tail moments in the combination group ( Fig. 4B and D, Supplementary file: Fig. S4B and D). To assess DNA damage and DDR pathway activation, we performed Western blotting to evaluate the expression of phosphorylated H2AX, TOPBP1, phosphorylated BRCA1, RAD51, 53BP1 and phosphorylated ATR (Fig. 4E). The results showed that the DNA damage marker γH2AX was remarkably increased when ...
Context 4
... of ESCC cells. Single-cell gel electrophoresis or comet assays were performed to assess the DNA damage induced by BI2536 and DDP. The results showed that both monotherapy and combination therapy could cause DNA strand breaks, as indicated by the higher frequency of Olive tail moments in the combination group ( Fig. 4B and D, Supplementary file: Fig. S4B and D). To assess DNA damage and DDR pathway activation, we performed Western blotting to evaluate the expression of phosphorylated H2AX, TOPBP1, phosphorylated BRCA1, RAD51, 53BP1 and phosphorylated ATR (Fig. 4E). The results showed that the DNA damage marker γH2AX was remarkably increased when cells were treated with both BI2536 and DDP, ...
Context 5
... strand breaks, as indicated by the higher frequency of Olive tail moments in the combination group ( Fig. 4B and D, Supplementary file: Fig. S4B and D). To assess DNA damage and DDR pathway activation, we performed Western blotting to evaluate the expression of phosphorylated H2AX, TOPBP1, phosphorylated BRCA1, RAD51, 53BP1 and phosphorylated ATR (Fig. 4E). The results showed that the DNA damage marker γH2AX was remarkably increased when cells were treated with both BI2536 and DDP, while this DNA damage marker was hardly detected in the control and single drug groups. These findings indicated that the combination of the two drugs caused significant DNA damage to the ESCC cells. ...
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Hepatocyte pyroptosis has been shown to be involved in liver damage progression. Previously, we found that growth arrest-specific 5 (GAS5) is a regulator of hepatic stellate cell (HSC) activation. However, whether GAS5 plays a role in hepatocyte pyroptosis remains unclear. In this study, reduced GAS5 was shown in CCl4-treated mice and restoration o...
Citations
... 49 In another study, BI2536, a PLK1 kinase inhibitor, increased the chemosensitivity of the tumor to DDP by promoting the conversion of apoptosis to pyroptosis in esophageal squamous cell carcinoma. 50 Therefore, it is necessary to further explore the use of molecular targeted drugs for pyroptosis-based tumor therapy, and nanomaterials can further improve the therapeutic effect. ...
Pyroptosis, a pro-inflammatory and lytic programmed cell death pathway, possesses great potential for antitumor immunotherapy. By releasing cellular contents and a large number of pro-inflammatory factors, tumor cell pyroptosis can promote dendritic cell maturation, increase the intratumoral infiltration of cytotoxic T cells and natural killer cells, and reduce the number of immunosuppressive cells within the tumor. However, the efficient induction of pyroptosis and prevention of damage to normal tissues or cells is an urgent concern to be addressed. Recently, a wide variety of nanoplatforms have been designed to precisely trigger pyroptosis and activate the antitumor immune responses. This review provides an update on the progress in nanotechnology for enhancing pyroptosis-based tumor immunotherapy. Nanomaterials have shown great advantages in triggering pyroptosis by delivering pyroptosis initiators to tumors, increasing oxidative stress in tumor cells, and inducing intracellular osmotic pressure changes or ion imbalances. In addition, the challenges and future perspectives in this field are proposed to advance the clinical translation of pyroptosis-inducing nanomedicines.
... In a study, the PLK1 inhibitor BI2536 increased cell sensitivity to DDP, leading to increased pyroptosis that killed the cancer cells. Combining these two drugs offers a novel pharmacological strategy for the management of ESCC [118]. ...
... However, more lethal exposures to CP markedly activate JNK and p38 in several cancer cell types, including HeLa cells, neuroblastoma, prostate cancer, lung carcinoma, hepatoma, breast cancer, and colorectal adenocarcinoma cell lines, leading to cell cycle arrest, then bcl2 proapoptotic proteins bax and bak initiate mitochondria-dependent cell death via cytochrome c release and caspase 9/3-driven apoptosis [39, 116,123,142,164]. In several cancer cell types with high gasdermin E (GSDME) expression, CP treatment also led to pyroptosis [142], a highly inflammatory manifestation of programmed necrosis caused by gasdermin (including GSDME) cleavage by caspase 3, leading to GSDMN pore formation that perforates the cell membrane [218][219][220][221]. ERK1/2 expression and phosphorylation in cancer cells can also be significantly downregulated by CP [144,222,223]. ...
Cold plasma (CP) is an ionised gas containing excited molecules and ions, radicals, and free electrons, and which emits electric fields and UV radiation. CP is potently antimicrobial, and can be applied safely to biological tissue, birthing the field of plasma medicine. Reactive oxygen and nitrogen species (RONS) produced by CP affect biological processes directly or indirectly via the modification of cellular lipids, proteins, DNA, and intracellular signalling pathways. CP can be applied at lower levels for oxidative eustress to activate cell proliferation, motility, migration, and antioxidant production in normal cells, mainly potentiated by the unfolded protein response, the nuclear factor-erythroid factor 2-related factor 2 (Nrf2)-activated antioxidant response element, and the phosphoinositide 3-kinase/protein kinase B (PI3K/Akt) pathway, which also activates nuclear factor-kappa B (NFκB). At higher CP exposures, inactivation, apoptosis, and autophagy of malignant cells can occur via the degradation of the PI3K/Akt and mitogen-activated protein kinase (MAPK)-dependent and -independent activation of the master tumour suppressor p53, leading to caspase-mediated cell death. These opposing responses validate a hormesis approach to plasma medicine. Clinical applications of CP are becoming increasingly realised in wound healing, while clinical effectiveness in tumours is currently coming to light. This review will outline advances in plasma medicine and compare the main redox and intracellular signalling responses to CP in wound healing and cancer.
... So, the High GSDME expression may explain why MTUS1/ATIP1-mediated pyroptosis induction in HNSCC cells. Our previous study had verified that MTUS1/ATIP1 induced apoptosis (9,14), So pyroptosis maybe a secondary necrosis after apoptosis (55,56,60). ...
We showed that microtubule-associated tumor suppressor gene (MTUS1/ATIP) downregulation correlated with poor survival in head and neck squamous cell carcinoma (HNSCC) patients and that MTUS1/ATIP1 was the most abundant isoform in HNSCC tissue. However, the location and function of MTUS1/ATIP1 have remain unclear. In this study, we confirmed that MTUS1/ATIP1 inhibited proliferation, growth and metastasis in HNSCC in cell- and patient-derived xenograft models in vitro and in vivo. MTUS1/ATIP1 localized in the outer mitochondrial membrane, influence the morphology, movement and metabolism of mitochondria and stimulated oxidative stress in HNSCC cells by directly interacting with MFN2. MTUS1/ATIP1 activated ROS, recruiting Bax to mitochondria, facilitating cytochrome c release to the cytosol to activate caspase-3, and inducing GSDME-dependent pyroptotic death in HNSCC cells. Our findings showed that MTUS1/ATIP1 localized in the outer mitochondrial membrane in HNSCC cells and mediated anticancer effects through ROS-induced pyroptosis, which may provide a novel therapeutic strategy for HNSCC treatment.
... Earlier studies identified Gasdermin D (GSDMD), a pyroptosis executor, which is cleaved in immunocytes following activation by caspase-1 and caspase-11/4/5 [6,7]. Recently, Gasdermin E (GSDME), another member of the pyroptosis family, was found to trigger pyroptosis in a variety of cancer cells [8][9][10][11]. Unlike GSDMD, activated caspase-3 cleaved GSDME, producing GSDME-N fragments that form pores within membranes, resulting in pyroptosis [12]. However, in the majority of cancer cells, including those from the stomach, breast, and colon, GSDME is silenced as a result of promoter DNA hypermethylation [13]. ...
Pyroptosis, a novel type of programmed cell death (PCD), which provides a feasible therapeutic option for the treatment of tumors. However, due to the hypermethylation of the promoter, the critical protein Gasdermin E (GSDME) is lacking in the majority of cancer cells, which cannot start the pyroptosis process and leads to dissatisfactory therapeutic effects. Additionally, the quick clearance, systemic side effects, and low concentration at the tumor site of conventional pyroptosis reagents restrict their use in clinical cancer therapy. Here, we described a combination therapy that induces tumor cell pyroptosis via the use of ultrasound-targeted microbubble destruction (UTMD) in combination with DNA demethylation. The combined application of UTMD and hydralazine-loaded nanodroplets (HYD-NDs) can lead to the rapid release of HYD (a demethylation drug), which can cause the up-regulation of GSDME expression, and produce reactive oxygen species (ROS) by UTMD to cleave up-regulated GSDME, thereby inducing pyroptosis. HYD-NDs combined with ultrasound (US) group had the strongest tumor inhibition effect, and the tumor inhibition rate was 87.15% (HYD-NDs group: 51.41 ± 3.61%, NDs + US group: 32.73%±7.72%), indicating that the strategy had a more significant synergistic anti-tumor effect. In addition, as a new drug delivery carrier, HYD-NDs have great biosafety, tumor targeting, and ultrasound imaging performance. According to the results, the combined therapy reasonably regulated the process of tumor cell pyroptosis, which offered a new strategy for optimizing the therapy of GSDME-silenced solid tumors.
Supplementary Information
The online version contains supplementary material available at 10.1186/s12951-024-02453-0.
... However, more lethal exposures to CP markedly activate JNK and p38 in several cancer cell types, including HeLa cells, neuroblastoma, prostate cancer, lung carcinoma, hepatoma, breast cancer, and colorectal adenocarcinoma cell lines, leading to cell cycle arrest, then bcl2 proapoptotic proteins bax and bak initiate mitochondria-dependent cell death via cytochrome c release and caspase 9/3-driven apoptosis [101,108,127,150,204]. In several cancer cell types with high gasdermin E (GSDME) expression, CP treatment also led to pyroptosis [127]; a highly inflammatory manifestation of programmed necrosis caused by gasdermin (including GSDME) cleavage by caspase 3 leading to GSDMN pore formation that perforates the cell membrane [205][206][207][208]. ERK1/2 expression and phosphorylation in cancer cells can also be significantly downregulated by CP [129,209,210]. ...
Cold plasma (CP) is an ionised gas containing excited molecules and ions, radicals, free electrons, and emits electric fields and UV radiation. CP is potently antimicrobial and can be applied safely to biological tissue, birthing the field of plasma medicine. Reactive oxygen and nitrogen species (RONS) produced by CP affect biological processes directly or indirectly via the modification of cellular lipids, proteins, DNA, and intracellular signalling pathways. CP can be applied at lower levels for oxidative eustress to activate cell proliferation, motility, migration, and antioxidant production in normal cells, mainly potentiated by the unfolded protein response, Nrf2-activated antioxidant response element and PI3K/Akt pathway, which also activates NFκB. At higher CP exposures, inactivation, apoptosis and autophagy of malignant cells can occur via degradation of PI3K/Akt and MAPK-dependent and -independent activation of the master tumour suppressor p53, leading to caspase-mediated cell death. These opposing responses validate a hormesis approach to plasma medicine. Clinical applications of CP are becoming increasingly realised in wound healing, while clinical effectiveness in tumours is currently coming to light. This review will outline advances in plasma medicine and compare the main redox and intracellular signalling responses to CP in wound healing and cancer.
... Additionally, it was found that the administration of BI2536/DDP could trigger the cleavage of caspase-3 and increase the level of GSDME in the cytoplasm, leading to pyroptosis in ESCC cells. Moreover, it was confirmed that the Bax/caspase-3/GSDME axis was responsible for pyroptosis induced by BI2536/DDP (Wu et al., 2019). ...
... Its mechanism of reducing PLK1 activity is mainly by targeting the ATP binding domain [19,20]. BI 2536 also exerts therapeutic effects on adrenocortical carcinoma by disrupting centrosome homeostasis and enhances the sensitivity of esophageal cancer to cisplatin by inducing apoptosis [21,22]. The combination of BI 2536 and cisplatin shows high anti-tumor activity in gastric cancer [23]. ...
Background
Polo-like kinase 1 (PLK1) is a critical therapeutic target in the treatment of head and neck squamous cell carcinoma (HNSCC). The objective of this study was to investigate the therapeutic effect of the combination of BI 2536, a PLK1 inhibitor, and erastin, a ferroptosis inducer, in HNSCC.
Methods
The proliferation, invasion, and migration abilities of Tu177 and FaDu cells upon exposure to BI 2536 and erastin, used in combination or alone, were tested. Fe²⁺, glutathione (GSH), and malondialdehyde (MDA) detection kits were used to determine whether the addition of BI 2536 enhanced the accumulation of Fe²⁺ and MDA, along with the depletion of GSH. Quantitative real-time PCR, western blot analyses were performed to investigate whether BI 2536 further altered the mRNA and expression level of ferroptosis genes. Furthermore, si PLK1 was used to investigate whether targeting PLK1 gene promoted erastin-induced ferroptosis.
Results
The combination of BI 2536 and erastin exerted a stronger cytotoxicity than treatment with a single agent. Compared with erastin treatment alone, the combination of BI 2536 and erastin lowered the ability of tumor cells to self-clone, invade, and migrate. BI 2536 enhanced the accumulation of Fe²⁺ and MDA, and the depletion of GSH. BI 2536 increased erastin-induced changes in ferroptosis-related gene mRNA and expression. Importantly, targeting PKL1 enhanced the anti-cancer effect of erastin.
Conclusion
BI 2536 enhanced the sensitivity of HNSCC cells to erastin, which provides a new perspective for cancer treatment.
... In vitro and in vivo tests demonstrated that BI-2536 inhibited the proliferation of osteosarcoma cell lines 74,75 . What's more, BI-2536 was reported to enhance the effects of various conventional chemotherapeutic agents [76][77][78] . Hence, for patients with the low risk scores, BI-2536 is a potentially potent complement to chemotherapy or neoadjuvant chemotherapy. ...
Osteosarcoma is generally considered a cold tumor and is characterized by epigenetic alterations. Although tumor cells are surrounded by many immune cells such as macrophages, T cells may be suppressed, be inactivated, or not be presented due to various mechanisms, which usually results in poor prognosis and insensitivity to immunotherapy. Immunotherapy is considered a promising anti-cancer therapy in osteosarcoma but requires more research, but osteosarcoma does not currently respond well to this therapy. The cancer immunity cycle (CIC) is essential for anti-tumor immunity, and is epigenetically regulated. Therefore, it is possible to modulate the immune microenvironment of osteosarcoma by targeting epigenetic factors. In this study, we explored the correlation between epigenetic modulation and CIC in osteosarcoma through bioinformatic methods. Based on the RNA data from TARGET and GSE21257 cohorts, we identified epigenetic related subtypes by NMF clustering and constructed a clinical prognostic model by the LASSO algorithm. ESTIMATE, Cibersort, and xCell algorithms were applied to analyze the tumor microenvironment. Based on eight epigenetic biomarkers (SFMBT2, SP140, CBX5, HMGN2, SMARCA4, PSIP1, ACTR6, and CHD2), two subtypes were identified, and they are mainly distinguished by immune response and cell cycle regulation. After excluding ACTR6 by LASSO regression, the prognostic model was established and it exhibited good predictive efficacy. The risk score showed a strong correlation with the tumor microenvironment, drug sensitivity and many immune checkpoints. In summary, our study sheds a new light on the CIC-related epigenetic modulation mechanism of osteosarcoma and helps search for potential drugs for osteosarcoma treatment.
... GSDME-mediated pyroptosis may increase the sensitivity of platinum by promoting T cell infiltration [42]. The efficacy of platinum could also be enhanced by combination with BI2536, a PLK1 kinase inhibitor, by inducing pyroptosis and affecting DNA damage repair [43]. Some noncoding RNAs can also sensitize chemoresistant cancer cells to cisplatin by activating pyroptosis [44]. ...
Pyroptosis is an active cell death process mediated by gasdermin family proteins including Gasdermin A (GSDMA), Gasdermin B (GSDMB), Gasdermin C (GSDMC), Gasdermin D (GSDMD), Gasdermin E (GSDME, DFNA5), and DFNB59. Emerging evidences have shown that pyroptosis contributes to many pulmonary diseases, especially lung cancer, and pneumonia. The exact roles of pyroptosis and gasdermin family proteins are tremendously intricate. Besides, there are evidences that pyroptosis contributes to these respiratory diseases. However, it often plays a dual role in these diseases which is a cause for concern and makes it difficult for clinical translation. This review will focus on the multifaceted roles of pyroptosis in respiratory diseases.
