Probing the Mechanism of Action of Cry41Aa on HepG2 Through the Establishment of a Resistant Subline

Overview

Journal Toxins (Basel)

Publisher MDPI

Specialty Toxicology

Date 2022 May 27

PMID 35622566

Authors

Wided Souissi

Tweedie Alistair

Barbara Domanska

Eva Fortea

Michelle J West

Jean-Louis Schwartz

Neil Crickmore

Affiliations

Soon will be listed here.

Abstract

Cry41Aa, also called parasporin-3, belongs to a group of toxins from the entomopathogenic bacterium that show activity against human cancer cells. Cry41Aa exhibits preferential cytocidal activity towards HL-60 (human promyelocytic leukaemia cells) and HepG2 (human liver cancer cells) cell lines after being proteolytically activated. To better understand the mechanism of action of Cry41Aa, we evolved resistance in HepG2 cells through repeated exposure to increasing doses of the toxin. Concentrations of Cry41Aa that killed over 50% of the parental HepG2 cells had no significant effect on the viability of the resistant cells and did not induce either pore formation or p38 phosphorylation (both characteristic features of pore-forming toxins). Preliminary RNA sequencing data identified AQP9 as a potential mediator of resistance, but extensive investigations failed to show a causal link and did not support an enhanced cell repair process as the resistance mechanism.

References

Kuppen P, Schuitemaker H, van t Veer L, de Bruijn E, Van Oosterom A, Schrier P . cis-diamminedichloroplatinum(II)-resistant sublines derived from two human ovarian tumor cell lines. Cancer Res. 1988; 48(12):3355-9. View

Los F, Kao C, Smitham J, McDonald K, Ha C, Peixoto C . RAB-5- and RAB-11-dependent vesicle-trafficking pathways are required for plasma membrane repair after attack by bacterial pore-forming toxin. Cell Host Microbe. 2011; 9(2):147-57. PMC: 3057397. DOI: 10.1016/j.chom.2011.01.005. View

Huffman D, Abrami L, Sasik R, Corbeil J, van der Goot F, Aroian R . Mitogen-activated protein kinase pathways defend against bacterial pore-forming toxins. Proc Natl Acad Sci U S A. 2004; 101(30):10995-1000. PMC: 503732. DOI: 10.1073/pnas.0404073101. View

Okamura M, Shimada J, Sakagami H . Comparative analysis of cell death induction by cisplatin and 5-FU in human oral squamous and hepatocellular carcinoma cell lines. Anticancer Res. 2008; 28(1A):253-9. View

Keyel P, Loultcheva L, Roth R, Salter R, Watkins S, Yokoyama W . Streptolysin O clearance through sequestration into blebs that bud passively from the plasma membrane. J Cell Sci. 2011; 124(Pt 14):2414-23. PMC: 3124372. DOI: 10.1242/jcs.076182. View

Ferre J, Van Rie J . Biochemistry and genetics of insect resistance to Bacillus thuringiensis. Annu Rev Entomol. 2001; 47:501-33. DOI: 10.1146/annurev.ento.47.091201.145234. View

Chan J, Chu A, Fung K . Inhibition of P-glycoprotein expression and reversal of drug resistance of human hepatoma HepG2 cells by multidrug resistance gene (mdr1) antisense RNA. Life Sci. 2000; 67(17):2117-24. DOI: 10.1016/s0024-3205(00)00798-0. View

Domanska B, Fortea E, West M, Schwartz J, Crickmore N . The role of membrane-bound metal ions in toxicity of a human cancer cell-active pore-forming toxin Cry41Aa from Bacillus thuringiensis. Toxicon. 2019; 167:123-133. DOI: 10.1016/j.toxicon.2019.06.003. View

Peyronnet O, Nieman B, Genereux F, Vachon V, Laprade R, Schwartz J . Estimation of the radius of the pores formed by the Bacillus thuringiensis Cry1C delta-endotoxin in planar lipid bilayers. Biochim Biophys Acta. 2002; 1567(1-2):113-22. DOI: 10.1016/s0005-2736(02)00605-3. View

10.

Endo H, Azuma M, Adegawa S, Kikuta S, Sato R . Water influx via aquaporin directly determines necrotic cell death induced by the Bacillus thuringiensis Cry toxin. FEBS Lett. 2016; 591(1):56-64. DOI: 10.1002/1873-3468.12506. View

11.

Ma G, Roberts H, Sarjan M, Featherstone N, Lahnstein J, Akhurst R . Is the mature endotoxin Cry1Ac from Bacillus thuringiensis inactivated by a coagulation reaction in the gut lumen of resistant Helicoverpa armigera larvae?. Insect Biochem Mol Biol. 2005; 35(7):729-39. DOI: 10.1016/j.ibmb.2005.02.011. View

12.

Luqmani Y . Mechanisms of drug resistance in cancer chemotherapy. Med Princ Pract. 2005; 14 Suppl 1:35-48. DOI: 10.1159/000086183. View

13.

Romero M, Keyel M, Shi G, Bhattacharjee P, Roth R, Heuser J . Intrinsic repair protects cells from pore-forming toxins by microvesicle shedding. Cell Death Differ. 2017; 24(5):798-808. PMC: 5423106. DOI: 10.1038/cdd.2017.11. View

14.

Xiao Y, Liu K, Zhang D, Gong L, He F, Soberon M . Resistance to Bacillus thuringiensis Mediated by an ABC Transporter Mutation Increases Susceptibility to Toxins from Other Bacteria in an Invasive Insect. PLoS Pathog. 2016; 12(2):e1005450. PMC: 4752494. DOI: 10.1371/journal.ppat.1005450. View

15.

Carroll J, Ellar D . Analysis of the large aqueous pores produced by a Bacillus thuringiensis protein insecticide in Manduca sexta midgut-brush-border-membrane vesicles. Eur J Biochem. 1997; 245(3):797-804. DOI: 10.1111/j.1432-1033.1997.00797.x. View

16.

Husmann M, Dersch K, Bobkiewicz W, Beckmann E, Veerachato G, Bhakdi S . Differential role of p38 mitogen activated protein kinase for cellular recovery from attack by pore-forming S. aureus alpha-toxin or streptolysin O. Biochem Biophys Res Commun. 2006; 344(4):1128-34. DOI: 10.1016/j.bbrc.2006.03.241. View

17.

Cooper S, McNeil P . Membrane Repair: Mechanisms and Pathophysiology. Physiol Rev. 2015; 95(4):1205-40. PMC: 4600952. DOI: 10.1152/physrev.00037.2014. View

18.

Gunning R, Dang H, Kemp F, Nicholson I, Moores G . New resistance mechanism in Helicoverpa armigera threatens transgenic crops expressing Bacillus thuringiensis Cry1Ac toxin. Appl Environ Microbiol. 2005; 71(5):2558-63. PMC: 1087549. DOI: 10.1128/AEM.71.5.2558-2563.2005. View

19.

Zhai B, Shao Z, Zhao C, Hu K, Wu F . Development and characterization of multidrug resistant human hepatocarcinoma cell line in nude mice. World J Gastroenterol. 2006; 12(41):6614-9. PMC: 4125665. DOI: 10.3748/wjg.v12.i41.6614. View

20.

Oppert B, Kramer K, Beeman R, Johnson D, McGaughey W . Proteinase-mediated insect resistance to Bacillus thuringiensis toxins. J Biol Chem. 1997; 272(38):23473-6. DOI: 10.1074/jbc.272.38.23473. View