Cell and Tissue BiologyCell and Tissue BiologyЦитология0041-37713034-6061The Russian Academy of Sciences66961510.31857/S0041377124020071RJVGQVArticlesСтатьиResearch ArticleAlpha-tocopheryl succinate induces ER stress, disregulates lipid metabolism and leads to apoptosis in normal and tumorous cell lines of epidermal originАльфа-токоферилсукцинат индуцирует стресс ЭПР, нарушение метаболизма липидов и апоптоз в культуре нормальных и опухолевых клеток эпидермального происхожденияSavitskayaM. A.СавицкаяМ. А.
Russian Federation

Department of Cell Biology and Histology

Кафедра клеточной биологии и гистологии

nakomis@mail.ruZakharovI. I.ЗахаровИ. И.
Russian Federation

Department of Cell Biology and Histology

Кафедра клеточной биологии и гистологии

nakomis@mail.ruSaidovaА. А.СаидоваА. А.
Russian Federation

Department of Cell Biology and Histology

Кафедра клеточной биологии и гистологии

nakomis@mail.ruSmirnovaЕ. А.СмирноваЕ. А.
Russian Federation

Department of Cell Biology and Histology

Кафедра клеточной биологии и гистологии

nakomis@mail.ruOnishchenkoG. E.ОнищенкоГ. Е.
Russian Federation

Department of Cell Biology and Histology

Кафедра клеточной биологии и гистологии

nakomis@mail.ruLomonosov Moscow State UniversityМосковский государственный университет им. М.В. Ломоносова1503202466217318727022025Copyright ©; 2024, Russian Academy of SciencesCopyright ©; 2024, Российская академия наук2024Russian Academy of SciencesРоссийская академия наукhttps://vietnamjournal.ru/0041-3771/article/view/669615

Vitamin E succinate (VES, α-tocopheryl succinate), is a potential antitumor agent known to selectively affect the mitochondria of tumor cells. However, the data on the proapoptotic mechanism of action of VES are unclear, and the effect of VES on normal, non-tumorigenic cells has not been fully investigated. Previously, we showed that VES induces apoptosis via the mitochondrial pathway in A431 human epidermoid carcinoma cells. The goal of this work is to investigate the effect of VES on non-tumorigenic cells and to reveal commonalities and differences in pathways activated in normal and tumorous cells. To achieve this, we studied how VES affects such organelles as the ER and the Golgi apparatus, analyzed the expression of ER stress-associated genes, and also assessed the ROS content and the accumulation of lipid droplets in A431 human epidermoid carcinoma cells and HaCaT immortalized human keratinocytes. We show that in both cell lines there are signs of ER stress, the amount of ROS and lipid droplets increases, as does the number of apoptotic cells. At the same time, the key difference in the mechanisms apoptotic cell death induction in A431 and HaCaT cells treated with VES lies in the reaction of mitochondria: in A431 cells, apoptotic cell death is triggered via the mitochondrial pathway, while HaCaT cells initiate apoptosis without involving mitochondria. Thus, the targets of VES in normal and tumor cells may differ and can possibly complement each other during apoptosis induction.

Сукцинат витамина Е (СВЕ) – потенциальный противоопухолевый агент, известный своим направленным воздействием на митохондрии опухолевых клеток. Однако данные о проапоптозном механизме действия СВЕ неоднозначны, а воздействие СВЕ на нормальные нетуморогенные клетки изучено недостаточно полно. Ранее удалось показать индукцию апоптоза по митохондриальному механизму при действии СВЕ на клетки эпидермоидной карциномы человека А431. Цель нашей работы – исследовать влияние СВЕ на нетуморогенные клетки и выявить общие механизмы, которые характерны как для нормальных, так и для опухолевых клеток, и механизмы, проявляющиеся только в одной из категорий клеток. Для достижения этой цели изучали действие СВЕ на такие органеллы, как эндоплазматический ретикулум (ЭПР) и аппарат Гольджи, анализировали экспрессию генов, связанных со стрессом ЭПР, а также оценивали содержание АФК и накопление липидных капель в цитоплазме в клетках эпидермоидной карциномы человека А431 и иммортализованных кератиноцитах человека НаСаТ. Показано, что в клетках обеих линий присутствуют признаки стресса ЭПР, возрастает содержание АФК и липидных включений, увеличивается число апоптотических клеток. При этом ключевое различие механизмов индукции апоптотической гибели клеток А431 и НаСаТ при действии СВЕ лежит в реакции митохондрий: в клетках А431 запуск апоптотической гибели осуществляется по митохондриальному механизму, в то время как клетки линии HaCaT вступают в апоптоз без участия митохондрий. Таким образом, мишени воздействия СВЕ на нормальные и опухолевые клетки могут различаться и, возможно, способны дополнять друг друга при индукции апоптоза.

α-tocopheryl succinateER stressapoptosislipid inclusionsROSα-токоферилсукцинатстресс ЭПРапоптозлипидные включенияАФКПравительство РФThe Russian Government121032300098-5Савицкая М.А., Вильданова М.С., Кисурина-Евгеньева О.П., Смирнова Е.А., Онищенко Г.Е. 2012. Митохондриальный путь апоптоза в клетках эпидермоидной карциномы человека А431 при действии α-токоферилсукцината. ActaNaturae. Т. 4. С. 93. (Savitskaya M.A., Vildanova M.S., Kisurina-Evgenieva O.P., Smirnova E.A., Onischenko G.E. 2012. Mitochondrial pathway of α-tocopheryl succinate-induced apoptosis in human epidermoid carcinoma A431 cells. ActaNaturae. V. 4. P. 88.)Савицкая М.А., Онищенко Г.Е. 2016. α-Токоферилсукцинат влияет на жизнеспособность, пролиферацию и дифференцировку опухолевых клеток. Биохимия. Т. 81. № 8. С. 1036. (Savitskaya M.A., Onischenko G.E. 2016. α-Tocopherylsuccinate affects malignant cell viability, proliferation, and differentiation. Biochemistry (Moscow). V. 8. P. 806.)Badamchian M., Spangelo B.L., Bao Y., Hagiwara Y., Hagiwara H., Ueyama H., Goldstein A.L. 1994. Isolation of a vitamin E analog from a green barley leaf extract that stimulates release of prolactin and growth hormone from rat anterior pituitary cells in vitro. J. NutrBiochem. V. 5. P. 145. https://doi.org/10.1016/0955-2863(94)90086-8Bjelakovic G., Nikolova D., Simonetti R.G., Gluud C. 2004. Antioxidant supplements for prevention of gastrointestinal cancers: a systematic review and meta-analysis. Lancet. V. 364. P. 1219.Bommiasamy H., Back S.H., Fagone P., Lee K., Meshinchi S., Vink E., Sriburi R., Frank M., Jackowski S., Kaufman R.J., Brewer J.W. 2009. ATF6-alpha induces XBP1-independent expansion of the endoplasmic reticulum. J. Cell Sci. V. 122. Pt. 10. P. 1626. https://doi.org/10.1242/jcs.045625Boren J., Brindle K.M. 2012. Apoptosis-induced mitochondrial dysfunction causes cytoplasmic lipid droplet formation. Cell Death Differ. V. 19. P. 1561. https://doi.org/10.1038/cdd.2012.34Delikatny E.J., Cooper W.A., Brammah S., Sathasivam N., Rideout D.C. 2002. Nuclear magnetic resonance-visible lipids induced by cationic lipophilic chemotherapeutic agents are accompanied by increased lipid droplet formation and damaged mitochondria. Cancer Res. V. 62. P. 1394.Dong L.F., Jameson V.J., Tilly D., Cerny J., Mahdavian E. 2011. Mitochondrial targeting of vitamin E succinate enhances its pro-apoptotic and anti-cancer activity via mitochondrial complex II. J. Biol. Chem. V. 286. P. 3717.Dong L.F., Low P., Dyason J.C., Wang X.F., Prochazka L., Witting P.K., Freeman R., Swettenham E., Valis K., Liu J., Zobalova R., Turanek J., Spitz D.R., Domann F.E., Scheffler I.E., Ralph S.J., Neuzil J. 2008. Alpha-tocopheryl succinate induces apoptosis by targeting ubiquinone-binding sites in mitochondrial respiratory complex II. Oncogene. V. 27. P. 4324.Dos Santos G.A., Abreu e Lima R.S., Pestana C.R., Lima A.S., Scheucher P.S., Thomé C.H., Gimenes-Teixeira H.L., Santana-Lemos B.A., Lucena-Araujo A.R., Rodrigues F.P., Nasr R., Uyemura S.A., Falcão R.P., de Thé H., Pandolfi P.P. et al. 2012. (+)α-Tocopheryl succinate inhibits the mitochondrial respiratory chain complex I and is as effective as arsenic trioxide or ATRA against acute promyelocytic leukemia in vivo. Leukemia. V. 26. P. 451.Gao F.F., Quan J.H., Lee M.A., Ye W., Yuk J.M., Cha G.H., Choi I.W., Lee Y.H. 2021. Trichomonasvaginalis induces apoptosis via ROS and ER stress response through ER-mitochondria crosstalk in SiHa cells. Parasit. Vectors. V. 14. P. 603. https://doi.org/10.1186/s13071-021-05098-2Gorman A.M., Healy S.J., Jäger R., Samali A. 2012. Stress management at the ER: regulators of ER stress-induced apoptosis. Pharmacol. Ther. V. 134. P. 306. https://doi.org/10.1016/j.pharmthera.2012.02.003.Gruber J., Staniek K., Krewenka C., Moldzio R., Patel A., Böhmdorfer S., Rosenau T., Gille L. 2014. Tocopheramine succinate and tocopheryl succinate: mechanism of mitochondrial inhibition and superoxide radical production. Bioorg. Med. Chem. V. 22. P. 684.Hakumäki J.M., Poptani H., Sandmair A.M., Ylä-Herttuala S., Kauppinen R.A. 1999. 1H MRS detects polyunsaturated fatty acid accumulation during gene therapy of glioma: implications for the in vivo detection of apoptosis. Nat. Med. V. 5. P. 1323. https://doi.org/10.1038/15279. PMID: 10546002Hapala I., Marza E., Ferreira T. 2011. Is fat so bad? Modulation of endoplasmic reticulum stress by lipid droplet formation. Biol. Cell. V. 103. P. 271. https://doi.org/10.1042/BC20100144. PMID: 21729000Hitomi J., Katayama T., Eguchi Y., Kudo T., Taniguchi M., Koyama Y., Manabe T., Yamagishi S., Bando Y., Imaizumi K., Tsujimoto Y., Tohyama M. 2004. Involvement of caspase-4 in endoplasmic reticulum stress-induced apoptosis and Abeta-induced cell death. J. Cell Biol. V. 165. P. 347. https://doi.org/10.1083/jcb.200310015Huang X., Li L., Zhang L., Zhang Z., Wang X., Zhang X., Hou L., Wu K. 2013. Crosstalk between endoplasmic reticulum stress and oxidative stress in apoptosis induced by α-tocopheryl succinate in human gastric carcinoma cells. Br. J. Nutr. V. 109. P. 727. https://doi.org/10.1017/S0007114512001882.Huang X., Zhang Z., Jia L., Zhao Y., Zhang X., Wu K. 2010. Endoplasmic reticulum stress contributes to vitamin E succinate-induced apoptosis in human gastric cancer SGC-7901 cells. Cancer Lett. V. 296. P. 123. https://doi.org/10.1016/j.canlet.2010.04.002Israel K., Yu W., Sanders B.G., Kline K. 2000. Vitamin E succinate induces apoptosis in human prostate cancer cells: role for Fas in vitamin E succinate-triggered apoptosis. Nutr. Cancer. V. 36. P. 90.Jarc E., Petan T. 2019. Lipid droplets and the management of cellular stress. Yale J. Biol. Med. V. 92. P. 435.Kogure K., Hama S., Manabe S., Tokumura A., Fukuzawa K. 2002. High cytotoxicity of alphatocopherylhemisuccinate to cancer cells is due to failure of their antioxidative defense systems. Cancer Lett. V. 186. P. 151.Majima D., Mitsuhashi R., Fukuta T., Tanaka T., Kogure K. 2019. Biological functions of α-tocopheryl succinate. J. Nutr. SciVitaminol. V. 65. P. S104. https://doi.org/10.3177/jnsv.65.S104. PMID: 31619606Neuzil J., Dyason J.C., Freeman R., Dong L.F., Prochazka L., Wang X.F., Scheffler I., Ralph S.J. 2007. Mitocans as anti-cancer agents targeting mitochondria: lessons from studies with vitamin E analogues, inhibitors of complex II. J. Bioenerg. Biomembr. V. 39. P. 65.Neuzil J., Weber T., Gellert N., Weber C. 2001. Selective cancer cell killing by alpha-tocopheryl succinate. Br. J. Cancer. V. 84. P. 87.Neuzil J., Zhao M., Ostermann G., Sticha M., Gellert N., Weber C., Eaton J.W., Brunk U.T. 2002. Alpha-tocopheryl succinate, an agent with in vivo anti-tumour activity, induces apoptosis by causing lysosomal instability. Biochem. J. V. 362. Pt. 3. P. 709. https://doi.org/10.1042/0264-6021:3620709Potashnikova D., Gladkikh A., Vorobjev I.A. 2015. Selection of superior reference genes’ combination for quantitative real-time PCR in B-cell lymphomas. Ann. Clin. Lab. V. 45. P. 64.Prochazka L., Dong L.F., Valis K., Freeman R., Ralph S.J., Turanek J., Neuzil J. 2010. alpha-Tocopheryl succinate causes mitochondrial permeabilization by preferential formation of Bak channels. Apoptosis. V. 15. P. 782.Qiu L.Z., Yue L.X., Ni Y.H., Zhou W., Huang C.S., Deng H.F., Wang N.N., Liu H., Liu X., Zhou Y.Q., Xiao C.R., Wang Y.G, Gao Y. 2021. Emodin-induced oxidative inhibition of mitochondrial function assists BiP/IRE1α/CHOP signaling-mediated ER-related apoptosis. Oxid. Med. Cell Longev. V. 2021. P. 8865813. https://doi.org/10.1155/2021/8865813Quintero M., Cabañas M.E., Arús C. 2010. 13C-labelling studies indicate compartmentalized synthesis of triacylglycerols in C6 rat glioma cells. Biochim. Biophys. Acta. V. 1801. P. 693. https://doi.org/10.1016/j.bbalip.2010.03.013Rauchová H., Vokurková M., Drahota Z. 2014. Inhibition of mitochondrial glycerol-3- phosphate dehydrogenase by α-tocopheryl succinate. Int. J. Biochem. Cell Biol. V. 53. P. 409.Rodriguez-Enriquez S., Marin-Hernandez A., Gallardo-Perez J.C., Carreno-Fuentes L., Moreno-Sanchez R. 2009. Targeting of cancer energy metabolism. Mol. Nutr. Food Res. V. 53. P. 29.Sriburi R., Jackowski S., Mori K., Brewer J.W. 2004. XBP1: a link between the unfolded protein response, lipid biosynthesis, and biogenesis of the endoplasmic reticulum. J. Cell Biol. V. 167. P. 35. https://doi.org/10.1083/jcb.200406136Verfaillie T., Garg A.D., Agostinis P. 2013. Targeting ER stress induced apoptosis and inflammation in cancer. Cancer Letters. V. 332. P. 249. https://doi.org/10.1016/j.canlet.2010.07.016Wang X.F., Witting P.K., Salvatore B.A., Neuzil J. 2005. Vitamin E analogs trigger apoptosis in HER2/erbB2-overexpressing breast cancer cells by signaling via the mitochondrial pathway. Biochem. Biophys. Res. Commun. V. 326. P. 282.Weber T., Dalen H., Andera L., Nègre-Salvayre A., Augé N., Sticha M., Lloret A., Terman A., Witting P.K., Higuchi M., Plasilova M.,Zivny J., Gellert N., Weber C., Neuzil J. 2003. Mitochondria play a central role in apoptosis induced by alpha-tocopheryl succinate, an agent with antineoplastic activity: comparison with receptor-mediated pro-apoptotic signaling. Biochemistry. V. 42. P. 4277.Wlodkowic D., Skommer J., McGuinness D., Hillier C., Darzynkiewicz Z. 2009. ER-Golgi network – a future target for anti-cancer therapy. Leuk Res. V. 33. P. 1440. https://doi.org/10.1016/j.leukres.2009.05.025Yang Y., Wang G., Wu W., Yao S., Han X., He D., He J., Zheng G., Zhao Y., Cai Z., Yu R. 2018. Camalexin Induces apoptosis via the ROS-ER stress-mitochondrial apoptosis pathway in AML Cells. Oxid. Med. Cell Longev. V. 2018: 7426950. https://doi.org/10.1155/2018/7426950Yu W., Sanders B.G., Kline K. 2003. RRR-alpha-tocopheryl succinate-induced apoptosis of human breast cancer cells involves Bax translocation to mitochondria. Cancer Res. V. 63. P. 2483.Zhao Y., Neuzil J., Wu K. 2009. Vitamin E analogues as mitochondria-targeting compounds: from the bench to the bedside? Mol. Nutr. Food Res. V. 53. P. 129.