Thiostrepton induces apoptotic cell death at the level of BCL-2/CED-9 in C. elegans

doi:10.21203/rs.3.rs-4847784/v1

Download PDF

Research Article

Thiostrepton induces apoptotic cell death at the level of BCL-2/CED-9 in C. elegans

https://doi.org/10.21203/rs.3.rs-4847784/v1

This work is licensed under a CC BY 4.0 License

You are reading this latest preprint version

Thiostrepton, a thiopeptide antibiotic, has been attracting increasing attention for its anti-proliferative and anti-cancer properties in various tested cell culture models. Extensive in vitro analysis has been conducted to understand its anti-cancer effect across multiple cancer types and cell lines, with numerous reports indicating that thiostrepton can inhibit cancer cell proliferation and tumor growth and induce apoptosis in vitro. On the other hand, the in vivo anti-tumor effect of thiostrepton remains elusive. In this study, we aimed to genetically and physiologically characterize the effects of thiostrepton on apoptosis induction in vivo using C. elegans. Our data demonstrate that thiostrepton induces apoptosis in C. elegans, and this apoptotic induction is independent of the genomic instability and is not related to P53 activity. Rather, the apoptotic cell death mediated by thiostrepton treatment occurs at the level of the BCL-2/CED-9 protein at the core apoptotic machinery. Furthermore, we have unlinked the high ROS (reactive oxygen species) induction reported in earlier in vitro studies from apoptosis induction upon thiostrepton treatment in C. elegans. Overall, our genetic data indicate that apoptosis induction mediated by thiostrepton occurs at the level of the BCL-2/CED-9 protein.

Cancer

Thiostrepton

Apoptosis

BCL-2/CED-9

C. elegans

ROS

In the last decade, a novel class of naturally occurring thiopeptide antibiotics characterized by their sulfur-containing heterocyclic rings has emerged. Thiopeptides are distinct from other natural products derived from peptides containing azoles due to a set of shared properties. Their defining property is a central six-membered ring containing nitrogen, which can exist in various oxidation states [1, 2]. Among these thiopeptide antibiotics is Thiostrepton (Thio), which was first derived from Streptomyces azureus in 1954 [3]. Historically, research on Thio focused on its antibacterial properties [4], highlighting its role as a potent blocker of prokaryotic translation by binding to 23S rRNA and the N-terminal domain of the ribosomal uL11 protein [5, 6]. The anti-cancer effect of thiostrepton was initially detected in a small molecule screening to hunt for FoxM1 inhibitors [7]. It is well established that the oncogenic transcription factor forkhead box M1 (FoxM1) is among the most upregulated genes in various carcinomas [8–10]. FoxM1 is a member of the forkhead/winged helix protein family that acts as a transcription factor in various physiological activities, including cell cycle progression, angiogenesis, and apoptosis [11]. Importantly, treating cancer cell lines with various thiazole antibiotics, such as Thio, specifically blocks FoxM1 transcriptional activity and expression without affecting the general transcriptional profile of the cancer cells [12]. Recent emerging data strongly suggest that the various thiazole antibiotics, such as siomycin A and thiostrepton, stabilize proteins like p53, Mcl-1, and p21 [12]. More importantly, it is now evident that the downregulation of FoxM1 mediated by Thio induces apoptotic cell death in various cancer cell lines, such as MCF7 breast carcinoma cells, lung cancer, melanoma, and colorectal cancer [13–16]. A more recent study has clearly linked FoxM1 downregulation by gene silencing or its inactivation using thiostrepton to cell cycle arrest and, subsequently, caspase-dependent apoptosis induction in the B-precursor acute lymphoblastic leukemia cells [11, 16]. Some studies have reported that Thio induces apoptosis through both p53-dependent and -independent mechanisms. Thio significantly reduces cancer cell migration, invasion, and angiogenesis, and hinders tumor growth and metastasis [15, 17]. While there have been extensive in vitro studies on the anti-cancer effect of Thio, the effect of Thio in an in vivo setting is yet to be characterized. In this study, we have used Caenorhabditis elegans (C. elegans) as an in vivo model system to investigate the effect of Thio on organismal development and its role in apoptosis induction.

Thiostrepton treatment induces apoptosis in C. elegans germline

Thiostrepton has been reported to have an anti-cancer effect on various cancer cell lines by blocking the transcriptional activity of FoxM1 and reducing its expression, leading to apoptosis induction [12]. To evaluate the effect of Thio in an in vivo setting, we used C. elegans to better understand the effect of Thio at an organismal level on cell death induction. To this end, we treated the larval L4 stage worms with various dosages of Thio (from 100 mM to 180 mM) for 24 hours and investigated the effect of this treatment on germline apoptosis induction. To score the apoptotic cells upon Thio treatment, we used the ced-1::gfp-expressing C. elegans line. CED-1 is a transmembrane protein that mediates apoptotic corpse engulfment, and its accumulation around the dying cells is a canonical marker to detect apoptotic corpses [18]. Treating ced-1::gfp worms with 150-mM and 180-mM doses of Thio resulted in a significant increase in apoptotic cell death in the germline of the developing C. elegans, confirmed by the accumulation of CED-1 protein around the dying cells (Fig.1a). Analyzing the pachytene region of the Thio-treated germline using DIC optics highlighted the morphological features of apoptotic cell death that differentiate the corpses from healthy non-dying cells. These features include a button-like appearance under the DIC microscope due to their engulfment by the germline sheath cells and changes in the refractive index of the dying cells (Fig.1b). Notably, the 180-μM concentration exhibited the highest number of apoptotic cells compared to the control (Fig.1a and 1b). Our result shows that 24-hour treatment with 180-mM Thio induced apoptosis in the C. elegans germline, while exposure of the developing worms for 24 hours to various concentrations of Thio (5–180 mM) did not result in developmental arrest nor changes in the gross anatomy (Suppl Fig.1).

Thiostrepton -induced germ cell apoptosis depends on the core apoptotic machinery

During C. elegans development, a total of 131 cells is eliminated by apoptosis as part of the worm’s normal development. Nearly all apoptotic cell deaths in C. elegans are indispensable to the core apoptotic machinery consisting of the BH3-only domain protein EGL-1, the BCL2-related protein CED-9, the worm’s ortholog of mammalian Apaf1 CED-4, and the sole C. elegans Caspase CED-3. The executive CED-3 Caspase is the most downstream component of the core apoptotic machinery in worms and is activated by its physical interaction with CED-4/Apaf1 [19-23]. To functionally characterize the molecular mechanism of Thio-induced apoptosis, we aimed to measure the effect of 180-mM Thio on germ cells in various apoptotic-deficient mutants. To this end, we treated ced-3(n2452) and ced-4(n1162) mutants carrying the CED-1::GFP marker with 180-mM Thio for 24-hours. ced-3(n2452) contains a deletion in ced-3, and ced-4(n1162) carries a point mutation that abolishes their proapoptotic function. As expected, in both mutants, Thio treatment failed to induce apoptosis in the pachytene-stage germ cells (Fig.2 a, b, and c). To test if Thio-induced apoptosis is dependent on CED-9/ BCL-2 protein, we measure apoptotic cell death in the germline of the ced-9 gain of function (GoF) strain upon exposure to Thio. Surprisingly, the apoptotic phenotype impost by Thio treatment was abolished in the ced-9 gain of function mutants (Fig.2 d and e).

Thiostrepton-induced germ cell apoptosis is independent of P53 function and the DNA damage response

In C. elegans, germ cell apoptosis occurs either as part of normal oogenesis and tissue hemostasis or upon the activation of the conserved DNA damage pathways mediated by the tumor suppressor protein P53 [24, 25]. C. elegans CEP-1, the ancestral homolog of the mammalian P53, transcriptionally induces the BH3-only protein egl-1 in response to genotoxic insult [26, 27]. To test if germ cell apoptosis induction upon Thio treatment was mediated by the activation of the conserved DNA damage pathway and CEP-1/P53, we scored the apoptotic germ cells upon treatment with 180-mM Thio in both cep-1(Ig12501), ced-1::gfp and egl-1(n1084n3082) mutants. cep-1(Ig12501) contains a loss-of-function mutation that hinders CEP-1 capacity to trigger apoptosis upon DNA damage, while egl-1 is a loss-of-function mutant that abolishes most DNA damage-induced cell death in the C. elegans germline. Interestingly, the cep-1 and egl-1 mutants exhibited apoptosis induction comparable to the wild-type strain, indicating that Thio induces apoptosis in the absence of functional cep-1/p53 (Fig.3 a, b, c, and d).

Although the apoptotic phenotype mediated by Thio treatment was independent of the P53 function, we aimed to further confirm that this treatment did not induce DNA damage response (DDR) nor genomic instability. To this end, we assessed the formation of RPA-1 foci in the germ cells upon Thio treatment. The RPA-1 protein is involved in various DNA metabolism, including DNA duplication, DNA damage repair, and the activation of the DNA repair pathway. The accumulation of RPA-1 foci on the chromatin is the canonical marker of the DNA breaks that trigger the activation of ATM/ATR protein kinases, leading to the activation of the canonical DNA damage checkpoint pathway [28]. To test if treating germline with Thio induces genomic instability, we exposed the SSM473 C. elegans transgenic strain carrying rpa-1::gfp with 180-mM Thio and measured the RPA-1 foci formation in the germ cells. As expected, we observed no significant differences in the number of RPA-1 foci upon treatment with Thio, confirming that Thio does not induce genomic instability (Suppl Fig 2a). To further confirm that Thio treatment did not activate the canonical DNA damage response pathway, we investigated the activity of Checkpoint Kinase 1 (CHK-1). Upon genotoxic stress, CHK-1 is phosphorylated on ser 345 through the activity of the ATM protein kinase. We therefore stained the germline of the Thio-treated worms with the antibody against CHK-1 p^{ser 345}. In line with our previous data, we observed no increase in CHK-1 phosphorylation upon Thio treatment (Suppl Fig. 2b). Altogether, our data indicate that the apoptotic phenotype induced by Thio is independent of P53 and unrelated to the genomic instability.

Thiostrepton induces apoptosis in the somatic cells of C. elegans

To test if Thio also induces somatic cell death and its effect is not limited to germ cells, we measured apoptosis induction in the developing worms. To this end, we took advantage of ced-1(e1735), a loss-of-function mutant strain that is defective in the elimination of apoptotic cells, making the apoptotic cells persist throughout the development. Interestingly, the Thio-treated L1 staged worms showed a significantly increased number of apoptotic cells in the pharyngeal region compared to the control (Suppl. Fig. 3a and b). These findings suggest that Thio treatment also induces somatic apoptosis and generally exerts its effect at the level of the core apoptotic machinery.

Thiostrepton treatment does not increase the level of radical oxygen species

It has previously been reported that Thio is capable of elevating intracellular radical oxygen species (ROS) levels invarious cell lines, and this elevation in ROS is believed to be linked to apoptotic induction [11, 29]. To test if Thio treatment induced ROS production in C. elegans and if an increase in ROS could be potentially linked to apoptosis induction, we quantified ROS levels in the wild-type worms with and without Thio treatment. Surprisingly, we observed no significant changes in ROS production upon Thio treatment (Fig. 4a and b). Our results demonstrate that a 24-hour Thio treatment does not increase intracellular ROS. To our surprise, Thio treatment reduced ROS elevation in the DMSO control sample to some degree. Several studies have shown that DMSO treatment leads to increased ROS production in cell cultures [30]. Additionally, we evaluated the expression level of the member of the Glutathione S Transferase GST-4, a key protein involved in the cellular response to oxidative stress, upon Thio treatment across multiple time points (6–24 hours). Previous reports have shown that gst-4::GFP is a robust reporter to measure cellular oxidative stress [31]. Although a slight increase in the GST-4::GFP signal was observed between 10 and 18 hours compared to the control, the overall elevation of the GFP signal was minimal and fluctuated stochastically across most of the time points. Altogether, there were no significant differences in intensities across the conditions and time points (Fig. 4c and d).

Thiostrepton -induced apoptosis is independent of FOXO/DAF-16

FoxM1 is an oncogenic transcriptional factor associated with various types of cancer [32-34] . There are over 100 members of forkhead transcriptional factors that are evolutionary and highly conserved from C. elegans to humans. Thio has been reported to inhibit FoxM1 transcriptional factor in a p53-dependent manner. To test if Thio-induced apoptosis was related to the C. elegans Foxo transcription factor, daf-16, we counted the number of apoptotic cells in the daf-16 mutant. To our surprise, treating daf-16 mutants with Thio induced apoptosis similar to the wild-type control (Fig. 4e and f), indicating that the apoptosis induction is independent of the daf-16/Foxo axis.

This study revealed the proapoptotic effect of thiostrepton (Thio) in C. elegans. We have demonstrated that the exposure of C. elegans to 180 µM of Thio induces caspase-dependent apoptosis at the level of CED-9/BCL-2. We have further shown that, unlike in mammalian cells, Thio-induced apoptosis in C. elegans is independent of the CEP-1/P53 function and unrelated to the DNA damage response. Also, we have clearly shown that the exposure of Thio to C. elegans does not lead to oxidative stress, although this finding contrasts with that reported in studies on human cells. Using various mutants of the core apoptotic machinery, we have shown that treating C. elegans with 180 µM of Thio results in apoptotic cell death. In this study, we used a well-customized model of C. elegans germline to characterize the apoptotic effect of Thio. Treating C. elegans with Thio at various concentrations for 24 hours leads to germ cell apoptosis that is dependent on the activity of the caspase CED-3. We have systematically investigated the proapoptotic effect of Thio in various mutants. As expected, the C. elegans counterpart of Apaf1, CED-4, is essential for Thio-induced apoptosis as a loss-of-function mutation in ced-4 abrogates the apoptotic phenotype. Interestingly, the apoptotic phenotype obverted with Thio treatment was dependent on the activity of the anti-apoptotic gene, CED-9/BCL-2. Thio failed to induce apoptosis in the germline of the CED-9 gains of function mutants, suggesting that the effect is Thio is most likely at the level of CED-9/ BCL-2 protein. This is in line with previous reports on the anti-cancer effect of Thio in melanoma cell lines. Treating melanoma cells with Thio induces apoptotic death that is inversely correlated with the antiapoptotic Mcl-1 expression level [35].

More recent studies have shown that Thio induces DNA damage response and activates the tumor suppressor protein P53 [15]. To understand the mechanism of Thio-induced apoptosis in an in vivo setting, we aimed to test if Thio could activate the DNA damage response and P53 induction. The antiproliferative and proapoptotic activity of Thio in vitro is well-established, and previous studies have reported its various modes of action. It has been shown that Thio physically interacts with and inhibits FoxM1. Thio treatment leads to the downregulation of FoxM1 at the RNA level, decreasing the proliferation rate in B-precursor acute lymphoblastic leukemia (B-pre-ALL) [1]. Other studies have suggested that Thio induces cell death by increasing the bax/bcl-2 ratio, a cell death switch, leading to the release of cytochrome c and activation of caspases [11].

On the other hand, treatment of worms carrying a loss-of-function mutation of cep-1/p53 with Thio did not influence the apoptotic response, indicating that Thio-induced apoptosis is independent of CEP-1/P53. Using cytological markers for the DNA damage response, we observed no signs of genomic instability upon Thio treatment, in contrast to human cells. We analyzed the chromatin-associated RPA-1 foci formation upon Thio treatment but observed no significant increase. Also, staining the isolated Thio-treated germline with an antibody specifically against the active form of CHK-1 protein was comparable with that of the wild-type. Altogether, our genetic and cytological data indicate that the apoptotic phenotype mediated by Thio is unrelated to the DNA damage response.

C. elegans Strains and maintenance:

C. elegans strains were maintained at 20°C on Nematode Growth Media (NGM) plates seeded with E. coli (OP50) bacteria as described previously [36]. C. elegans strains used in this study: Bristol N2 (wildtype), MD701: bcIs39 [lim-7p::ced-1::GFP + lin-15(+)], EPD077: cep-1(Ig12501);[lim-7p::ced-1::GFP + lin-15(+)], KX84: ced-3(n2452) IV; bcIs39 V, CB3203: ced-1(e1735)I, KX89: ced-4(n1162) III; bcIs39 [lim-7p::ced-1::GFP + lin-15(+)], SSM473: rpa-1(iow89[GFP11::rpa-1])II; iowSi8II; unc-119(ed3)III, MT4770: ced-9 (n1950) III, CF1038: daf-16 (mu86), MT8735: egl-1(n1084n3082), and CL2166: dvIs19 [(pAF15) gst-4p::gfp::nls].

Thiostrepton treatment:

The L4-stage worms were treated with a corresponding dosage of Thio dissolved in dimethyl sulfoxide (DMSO) in the liquid culture and were incubated for 24-hours. For the control, the corresponding DMSO concentration of the pair of experimental samples was used.

Scoring of apoptotic cells:

The apoptotic cells were visualized and scored using the Leica 3D-Thunder Imager. Apoptotic cells were identified as previously described by either the DIC optic or the presence of the CED-1::GFP [34]. Apoptotic cells of the pharyngeal region of the F1 generation (highly synchronized L1-staged larvae) of the Thio-treated worms were scored using DIC microscopy.

Immunostaining and microscopy:

Synchronized wild-type worms were grown until they reached the L4 stage and were treated with Thiostrepton and DMSO (control). Twenty-four hours later, germlines were extracted and immunostained as previously described [19, 37]. The primary and secondary antibodies used are as follows: Primary antibodies: Phospho-CHK1 (Ser345) Rabbit Monoclonal Antibody. Secondary antibodies used in this study: Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 568) (1:200). DAPI staining was used at a concentration of 0.5 μg/ml. The 3D-Thunder Imager was used to visualize the samples. The small Volume Computational Clearing (SVCC) method was used to increase the clarity of the images. Either a single z stack (~0.3 μm) or the maximum projection of multiple stacks was used to generate the images.

Analyses of oxidative stress:

To detect Reactive Oxygen Species (ROS), the CellROX Green staining kit was used (C10444, Invitrogen, USA). The L4 staged worms were treated with Thio (180 μM) for 24-hours. Adult worms were washed three times with M9 and were fixed for 30 minutes in 2% formaldehyde. Fixed samples were washed three times in PBS prior to staining. Samples were stained with 5 μM of the CellROX Green solution for 1 hour at room temperature and then washed three times with PBS. The stained samples were mounted for microscopic analysis. Images were analyzed using ImageJ.

Statistical analysis:

Statistical analyses were performed using GraphPad Prism Version 10.1.0 (264) (GraphPad Software, San Diego, CA, USA). To compare the mean values among the control and treated groups, a two-way analysis of variance (ANOVA) test was used. The calculated p-values were used as indicators of statistical significance. The threshold for statistical significance was set at p< 0.05, and the levels of statistical significance were designated as: *p<0.05, **p<0.01, ***p<0.001, and ****p<0.0001.

Acknowledgments

We would like to thank the members of the Pourkarimi Lab for their constructive comments and feedback throughout this work. We would also like to acknowledge the Caenorhabditis Genetics Center (CGC), funded by the NIH (National Institute of Health) Office of Research Infrastructure Programs (P40OD010440), for providing C. elegans strains. This work and the Pourkarimi Lab are funded by the Qatar Foundation and the College of Health and Life Sciences at Hamad Bin Khalifa University (HBKU).

Author Contributions

Conceptualization, investigation, methodology, visualization, writing of the original draft, AK.; conceptualization, investigation, T.A.A.; conceptualization, investigation, MI.; investigation, KP.; resources, validation, review, SU.; conceptualization, project administration, resources, supervision, validation, writing, review, and editing of the original manuscript, E.P. All authors have read and agreed to the published version of the manuscript.

Compliance with ethical standards

Conflict of interest: The authors declare no conflict of interest.

Kwok, J.M., et al., Thiostrepton selectively targets breast cancer cells through inhibition of forkhead box M1 expression. Molecular cancer therapeutics, 2008. 7(7): p. 2022-2032.
Just-Baringo, X., F. Albericio, and M. Álvarez, Thiopeptide antibiotics: retrospective and recent advances. Marine drugs, 2014. 12(1): p. 317-351.
Donovick, R., et al., Thiostrepton, a new antibiotic. I. In vitro studies. Antibiotics annual, 1955. 3: p. 554-559.
Bailly, C., The bacterial thiopeptide thiostrepton. An update of its mode of action, pharmacological properties and applications. European Journal of Pharmacology, 2022. 914: p. 174661.
Wang, S., et al., Target-oriented design and biosynthesis of thiostrepton-derived thiopeptide antibiotics with improved pharmaceutical properties. Organic Chemistry Frontiers, 2015. 2(2): p. 106-109.
Polikanov, Y.S., et al., The mechanisms of action of ribosome-targeting peptide antibiotics. Frontiers in molecular biosciences, 2018. 5: p. 48.
Radhakrishnan, S.K., et al., Identification of a chemical inhibitor of the oncogenic transcription factor forkhead box M1. Cancer research, 2006. 66(19): p. 9731-9735.
Bektas, N., et al., Tight correlation between expression of the Forkhead transcription factor FOXM1 and HER2 in human breast cancer. BMC cancer, 2008. 8: p. 1-9.
Garber, M.E., et al., Diversity of gene expression in adenocarcinoma of the lung. Proceedings of the National Academy of Sciences, 2001. 98(24): p. 13784-13789.
Van den Boom, J., et al., Characterization of gene expression profiles associated with glioma progression using oligonucleotide-based microarray analysis and real-time reverse transcription-polymerase chain reaction. The American journal of pathology, 2003. 163(3): p. 1033-1043.
Kuttikrishnan, S., et al., Thiostrepton inhibits growth and induces apoptosis by targeting FoxM1/SKP2/MTH1 axis in B-precursor acute lymphoblastic leukemia cells. Leukemia & Lymphoma, 2021. 62(13): p. 3170-3180.
Gartel, A.L., FoxM1 inhibitors as potential anticancer drugs. Expert opinion on therapeutic targets, 2008. 12(6): p. 663-665.
Ju, S., et al., Identification of thiostrepton as a novel therapeutic agent that targets human colon cancer stem cells. Cell death & disease, 2015. 6(7): p. e1801-e1801.
Kongsema, M., et al., Molecular mechanism of Forkhead box M1 inhibition by thiostrepton in breast cancer cells. Oncology Reports, 2019. 42(3): p. 953-962.
Jiang, L., et al., Targeting FoxM1 by thiostrepton inhibits growth and induces apoptosis of laryngeal squamous cell carcinoma. Journal of Cancer Research and Clinical Oncology, 2015. 141: p. 971-981.
Khan, I., et al., Nuclear FOXM1 drives chemoresistance in AML. Leukemia, 2017. 31(1): p. 251-255.
Pandit, B. and A.L. Gartel, Proteasome inhibitors induce p53-independent apoptosis in human cancer cells. The American journal of pathology, 2011. 178(1): p. 355-360.
Zhou, Z., E. Hartwieg, and H.R. Horvitz, CED-1 is a transmembrane receptor that mediates cell corpse engulfment in C. elegans. Cell, 2001. 104(1): p. 43-56.
Pourkarimi, E., S. Greiss, and A. Gartner, Evidence that CED-9/Bcl2 and CED-4/Apaf-1 localization is not consistent with the current model for C. elegans apoptosis induction. Cell Death & Differentiation, 2012. 19(3): p. 406-415.
Ellis, H.M. and H.R. Horvitz, Genetic control of programmed cell death in the nematode C. elegans. Cell, 1986. 44(6): p. 817-829.
Yuan, J. and H.R. Horvitz, The Caenorhabditis elegans cell death gene ced-4 encodes a novel protein and is expressed during the period of extensive programmed cell death. Development, 1992. 116(2): p. 309-320.
Hengartner, M.O., R. Ellis, and R. Horvitz, Caenorhabditis elegans gene ced-9 protects cells from programmed cell death. Nature, 1992. 356(6369): p. 494-499.
Izadi, M., T.A. Ali, and E. Pourkarimi, Over fifty years of life, death, and cannibalism: a historical recollection of apoptosis and autophagy. International journal of molecular sciences, 2021. 22(22): p. 12466.
Gartner, A., et al., A conserved checkpoint pathway mediates DNA damage–induced apoptosis and cell cycle arrest in C. elegans. Molecular cell, 2000. 5(3): p. 435-443.
Gumienny, T.L., et al., Genetic control of programmed cell death in the Caenorhabditis elegans hermaphrodite germline. Development, 1999. 126(5): p. 1011-1022.
Schumacher, B., et al., The C. elegans homolog of the p53 tumor suppressor is required for DNA damage-induced apoptosis. Current biology, 2001. 11(21): p. 1722-1727.
Derry, W.B., A.P. Putzke, and J.H. Rothman, Caenorhabditis elegans p53: role in apoptosis, meiosis, and stress resistance. Science, 2001. 294(5542): p. 591-595.
Zou, Y., et al., Functions of human replication protein A (RPA): from DNA replication to DNA damage and stress responses. Journal of cellular physiology, 2006. 208(2): p. 267-273.
Zhang, W., et al., Thiostrepton induces ferroptosis in pancreatic cancer cells through STAT3/GPX4 signalling. Cell death & disease, 2022. 13(7): p. 630.
Sadowska-Bartosz, I., et al., Dimethyl sulfoxide induces oxidative stress in the yeast Saccharomyces cerevisiae. FEMS yeast research, 2013. 13(8): p. 820-830.
Link, C.D. and C.J. Johnson, Reporter transgenes for study of oxidant stress in Caenorhabditis elegans, in Methods in enzymology. 2002, Elsevier. p. 497-505.
Halasi, M. and A.L. Gartel, FOX (M1) news—it is cancer. Molecular cancer therapeutics, 2013. 12(3): p. 245-254.
Nandi, D., et al. FoxM1: Repurposing an oncogene as a biomarker. in Seminars in Cancer Biology. 2018. Elsevier.
Prabhu, K.S., et al., Bortezomib exerts its anti-cancer activity through the regulation of Skp2/p53 axis in non-melanoma skin cancer cells and C. elegans. Cell Death Discovery, 2024. 10(1): p. 225.
Bhat, U.G., et al., Novel anticancer compounds induce apoptosis in melanoma cells. Cell cycle, 2008. 7(12): p. 1851-1855.
Brenner, S., The genetics of Caenorhabditis elegans. Genetics, 1974. 77(1): p. 71-94.
Izadi, M., et al., Tryptophanyl-tRNA synthetase-1 (WARS-1) depletion and high tryptophan concentration lead to genomic instability in Caenorhabditis elegans. Cell Death Discovery, 2024. 10(1): p. 165.

No competing interests reported.

SupplementalFigures.docx

Download PDF

Reviewers agreed at journal
12 Sep, 2024
Reviewers invited by journal
11 Sep, 2024
Editor assigned by journal
03 Aug, 2024
Submission checks completed at journal
03 Aug, 2024
First submitted to journal
02 Aug, 2024

You are reading this latest preprint version

Thiostrepton induces apoptotic cell death at the level of BCL-2/CED-9 in C. elegans

Status:

Version 1

Abstract

Figures

Introduction

Results

Discussion

Materials and Methods

Declarations

References

Additional Declarations

Supplementary Files

Status:

Version 1