Effect of  Mutation in the LLC1 Mouse Lewis Lung Adenocarcinoma Line on Sensitivity to Particle Radiotherapy

Gantsova, E.A.; Arutyunyan, I.V.; Soboleva, A.G.; Shakirova, K.M.; Kananykhina, E.Yu.; Balchir, D.V.; Vishnyakova, P.A.; Saburov, V.O.; Gordon, K.B.; Fatkhudinov, T.Kh.

doi:10.7868/S1998286025050204

PII

S19982860S0132342325050204-1

DOI

10.7868/S1998286025050204

Publication type

Article

Status

Published

Authors

E.A. Gantsova

Affiliation:
Research Institute of Molecular and Cellular Medicine, Patrice Lumumba Peoples' Friendship University of Russia
A.V. Avtsyn Research Institute of Human Morphology, P.G. Petrovsky National Research Center of Surgery

I.V. Arutyunyan

Affiliation:
Research Institute of Molecular and Cellular Medicine, Patrice Lumumba Peoples' Friendship University of Russia
A.V. Avtsyn Research Institute of Human Morphology, P.G. Petrovsky National Research Center of Surgery

A.G. Soboleva

Affiliation:
Research Institute of Molecular and Cellular Medicine, Patrice Lumumba Peoples' Friendship University of Russia
A.V. Avtsyn Research Institute of Human Morphology, P.G. Petrovsky National Research Center of Surgery

K.M. Shakirova

Affiliation:
Research Institute of Molecular and Cellular Medicine, Patrice Lumumba Peoples' Friendship University of Russia
N.I. Pirogov Russian National Research Medical University

E.Yu. Kananykhina

Affiliation: A.V. Avtsyn Research Institute of Human Morphology, P.G. Petrovsky National Research Center of Surgery

D.V. Balchir

Affiliation: Research Institute of Molecular and Cellular Medicine, Patrice Lumumba Peoples' Friendship University of Russia

P.A. Vishnyakova

Affiliation: Research Institute of Molecular and Cellular Medicine, Patrice Lumumba Peoples' Friendship University of Russia

V.O. Saburov

Affiliation: A.F. Tsyb Medical Radiological Research Center – Branch of the National Medical Research Center for Radiology, Ministry of Health of the Russian Federation

K.B. Gordon

Affiliation:
Research Institute of Molecular and Cellular Medicine, Patrice Lumumba Peoples' Friendship University of Russia
A.F. Tsyb Medical Radiological Research Center – Branch of the National Medical Research Center for Radiology, Ministry of Health of the Russi

T.Kh. Fatkhudinov

Affiliation:
Research Institute of Molecular and Cellular Medicine, Patrice Lumumba Peoples' Friendship University of Russia
A.V. Avtsyn Research Institute of Human Morphology, P.G. Petrovsky National Research Center of Surgery

Volume/ Edition

Volume 51 / Issue number 5

Pages

953-965

Abstract

Lung adenocarcinoma is a malignant tumor, which is the most common type of non-small cell lung cancer. Low efficiency of standard methods of treatment of lung adenocarcinoma with mutation of the tumor suppressor gene is a serious problem in clinical practice. Search and improvement of new therapeutic approaches to this disease remains an urgent task of modern biomedicine. The aim of the work was to create an model of lung cancer based on the LLC1 cell line with knockout of the gene to assess the sensitivity of mutant cells to various types of radiation therapy, including irradiation with photons, protons and neutrons. The main methods used were CRISPR/Cas9 genome editing technologies to obtain mutant clones, laser cell sorting, PCR analysis to confirm the deletion, as well as assessment of viability, proliferation (metabolic tests, marker expression), apoptosis induction (annexin V-PI method) and gene expression after cell irradiation with a dose of 2 Gy. As a result, heterozygous mutant lines LLC1-STK11-Mut were obtained. Cell irradiation revealed that in mutant cells, radio-induced growth stimulation persisted longer than in wild-type cells, and a significant increase in the proportion of late apoptotic and necrotic cells was observed. At the same time, expression temporarily decreased after irradiation, but quickly recovered in mutant cells, which indicates their higher radioresistance. Unlike wild-type cells, the expression level of the gene in mutant cells did not change significantly after irradiation. Thus, the mutation contributes to the formation of radioresistance in tumor cells by triggering various adaptation mechanisms. The obtained model can be used for further study of radioresistance and development of new approaches to the therapy of tumors with mutation.

Keywords

аденокарцинома легкого серин/треониновая киназа 11 (STK11) адронная терапия терапия частицами иммунотерапия рака

Date of publication

01.05.2025

Year of publication

2025

Number of purchasers

Views

References

1. Khozyainova A.A., Menyailo M.E., Tretyakova M.S., Bokova U.A., Korobeynikova A.A., Gerashchenko T.S., Rodionov E.O., Miller S.V., Denisov E.V. // RUDN J. Med. 2024. V. 28. P. 439–451. https://doi.org/10.22363/2313-0245-2024-28-4-439-451
2. Spella M., Stathopoulos G.T. // Cancers (Basel). 2021. V. 13. P. 384. https://doi.org/10.3390/cancers13030384
3. Xin S., Wen M., Tian Y., Dong H., Wan Z., Jiang S., Meng F., Xiong Y., Han Y. // BMC Cancer. 2025. V. 23. P. 66. https://doi.org/10.1186/S12957-025-03701-9
4. Schabath M.B., Welsh E.A., Fulp W.J., Chen L., Teer J.K., Thompson Z.J., Engel B.E., Xie M., Berglund A.E., Creelan B.C., Antonia S.J., Gray J.E., Eschrich S.A., Chen D.T., Cress W.D., Haura E.B., Beg A.A. // Oncogene. 2016. V. 35. P. 3209–3216. https://doi.org/10.1038/onc.2015.375
5. Pons-Tostivint E., Lugat A., Fontenau J.F., De-nis M.G., Bennouna J. // Cells. 2021. V. 10. P. 3129. https://doi.org/10.3390/cells10113129
6. Facchinetti F., Bluthgen M.V., Tergemina-Clain G., Faivre L., Pignon J.P., Planchard D., Remon J., Soria J.C., Lacroix L., Besse B. // Lung Cancer. 2017. V. 112. P. 62–68. https://doi.org/10.1016/j.lungcan.2017.06.016
7. Mazzaschi G., Leonetti A., Minari R., Gnetti L., Quaini F., Tiseo M., Facchinetti F. // Curr. Treat. Options Oncol. 2021. V. 22. P. 96. https://doi.org/10.1007/s11864-021-00891-8
8. Zhang C.Y., Sun H., Su J.W., Chen Y.Q., Zhang S.L., Zheng M.Y., Li Y.F., Huang J., Zhang C., Tai Z.X., Cai M., Zhang X.C., Su J., Xu C.R., Yan H.H., Chen H.J., Wu Y.L., Yang J.J. // Lung Cancer. 2023. V. 175. P. 68–78. https://doi.org/10.1016/j.lungcan.2022.11.016
9. Jumaniyazova E.D., Sentyabreva A.V., Kosyreva A.M., Lokhonina A.V. // RUDN J. Med. 2024. V. 28. P. 413–426. https://doi.org/10.22363/2313-0245-2024-28-4-413-426
10. Kiseleva V., Gordon K., Vishnyakova P., Gantsova E., Elchaninov A., Fatkhudinov T. // Life (Basel). 2022. V. 12. P. 2071. https://doi.org/10.3390/life12122071
11. Jumaniyazova E., Smyk D., Vishnyakova P., Fatkhudinov T., Gordon K. // Life (Basel). 2023. V. 13. P. 30. https://doi.org/10.3390/LIFE13010030
12. Mirjolet C., Nicol A., Limagne E., Mura C., Richard C., Morgand V., Rousseau M., Boidot R., Ghiringhelli F., Noel G., Burckel H. // Sci. Rep. 2021. V. 11. P. 13444. https://doi.org/10.1038/s41598-021-92942-1
13. Qu Y., Zhang B., Liu S., Zhang A., Wu T., Zhao Y. // Cell. Mol. Immunol. 2010. V. 7. P. 419–427. https://doi.org/10.1038/cmi.2010.45
14. Gordon K., Gulidov I., Fatkhudinov T., Koryakin S., Kaprin A. // Int. J. Part. Ther. 2022. V. 9. P. 59– 69. https://doi.org/10.14338/IJPT-22-00017
15. Stankevicius V., Kuodyte K., Schveigert D., Bulotiene D., Paulauskas T., Daniunaite K., Suziedelis K. // Oncol. Lett. 2017. V. 13. P. 4190–4200. https://doi.org/10.3892/OL.2017.5877
16. Fasola C.E., Jones J.C., Huang D.D., Le Q.T., Hoppe R.T., Donaldson S.S. // Int. J. Radiat. Oncol. Biol. Phys. 2013. V. 86. P. 930–935. https://doi.org/10.1016/J.IJROBP.2013.04.035
17. Schaub L., Harrabi S.B., Debus J. // Br. J. Radiol. 2020. V. 93. P. 20200183. https://doi.org/10.1259/bjr.20200183
18. Kuwahara Y., Mori M., Kitahara S., Fukumoto M., Ezaki T., Mori S., Echigo S., Ohkubo Y., Fukumoto M. // Cancer Med. 2014. V. 3. P. 310–321. https://doi.org/10.1002/CAM4.185
19. Mardynsky Y.S., Gulidov I.A., Ivanov S.A., Kaprin A.D., Gordon K.B. // Radiat. Risk. 2024. V. 33. P. 40–54. https://doi.org/10.21870/0131-3878-2024-33-1-40-54
20. Hardie D.G., Alessi D.R. // BMC Biol. 2013. V. 11. Р. 36. https://doi.org/10.1186/1741-7007-11-36
21. Pierce S.E., Granja J.M., Corces M.R., Brady J.J., Tsai M.K., Pierce A.B., Tang R., Chu P., Feldser D.M., Chang H.Y., Bassik M.C., Greenleaf W.J., Winslow M.M. // Nat. Cell Biol. 2021. V. 23. P. 915– 924. https://doi.org/10.1038/S41556-021-00728-4
22. Urruticoechea A., Smith I.E., Dowsett M. // J. Clin. Oncol. 2005. V. 23. P. 7212–7220. https://doi.org/10.1200/JCO.2005.07.501
23. Oh M., Tanaka T., Kobayashi M., Furukawa T., Mori T., Kudo T., Fujieda S., Fujibayashi Y. // Nucl. Med. Biol. 2009. V. 36. P. 419–426. https://doi.org/10.1016/J.NUCMEDBIO.2009.01.016
24. van Gogh M., Garzon J.F.G., Sahin D., Knopfova L., Benes P., Boyman O., Jurisica I., Borsig L. // Cancer Immunol. Res. 2023. V. 11. P. 1432–1444. https://doi.org/10.1158/2326-6066.cir-22-0912
25. Andrés-Sánchez N., Fisher D., Krasinska L. // J. Cell Sci. 2022. V. 135. Р. jcs258932. https://doi.org/10.1242/jcs.258932
26. Mrouj K., Andrés-Sánchez N., Dubra G., Singh P., Sobecki M., Chahar D., Al Ghoul E., Aznar A.B., Prieto S., Pirot N., Bernex F., Bordignon B., Hassen- Khodja C., Villalba M., Krasinska L., Fisher D. // Proc. Natl. Acad. Sci. USA. 2021. V. 118. P. e2026507118. https://doi.org/10.1073/pnas.2026507118
27. Zhang Y.P., Guo Z.Q., Cai X.T., Rong Z.X., Fang Y., Chen J.Q., Zhuang K.M., Ruan M.J., Ma S.C., Lin L.Y., Han D.D., Li Y.S., Wang Y.Y., Wang J., Cao C.H., Tang X.R., Xie Q.K., Chen Y., Lin Y., Le Tan J., Yu Z.H., Wu Z.N., Wei W., Zheng D.Y., Zeng Y.J., Ruan Y.C., Xu Z.P., Gu J.Z., Xiao L.S., Liu L., Guan J., Bai X., Wu D.H., Dong Z.Y. // Cancer Cell. 2025. V. 43. Р. 856–874.e9. https://doi.org/10.1016/j.ccell.2025.02.024
28. Jung S., Jiang L., Zhao J., Shultz L.D., Greiner D.L., Bae M., Li X., Ordikhani F., Kuai R., Joseph J., Kasinath V., Elmaleh D.R., Abdi R. // Bioeng. Transl. Med. 2023. V. 8. P. e10273. https://doi.org/10.1002/btm2.10273
29. Exposito F., Redrado M., Houry M., Hastings K., Molero-Abraham M., Lozano T., Solorzano J.L., Sanz-Ortega J., Adradas V., Amat R., Redin E., Leon S., Legarra N., Garcia J., Serrano D., Valencia K., Robles-Oteiza C., Foggetti G., Otegui N., Felip E., Lasarte J.J., Paz-Ares L., Zugazagoitia J., Politi K., Montuenga L., Calvo A. // Cancer Res. 2023. V. 83. P. 2513–2526. https://doi.org/10.1158/0008-5472.CAN-22-2900
30. Lázaro S., Lorz C., Enguita A.B., Seller I., Paramio J.M., Santos M.// Cancers (Basel). 2022. V. 14. P. 3671. https://doi.org/10.3390/cancers14153671
31. Stankevicius V., Kuodyte K., Schveigert D., Bulotiene D., Paulauskas T., Daniunaite K., Suziedelis K. // Oncol. Lett. 2017. V. 13. P. 4190–4200. https://doi.org/10.3892/ol.2017.5877
32. Vallabhaneni S., Liu J., Morel M., Wang J., De-Mayo F.J., Long W. // Mol. Oncol. 2022. V. 16. P. 1184–1199. https://doi.org/10.1002/1878-0261.13173
33. Long L.L., Ma S.C., Guo Z.Q., Zhang Y.P., Fan Z., Liu L.J., Liu L., Han D.D., Leng M.X., Wang J., Guo X.J., Le Tan J., Cai X.T., Lin Y., Pan X., Wu D.H., Bai X., Dong Z.Y. // Cancer Res. 2023. V. 83. P. 568–581. https://doi.org/10.1158/0008-5472.can-22-1740
34. Bai X., Guo Z.Q., Zhang Y.P., Fan Z.-Z., Liu L.J., Liu L., Long L.L., Ma S.C., Wang J., Fang Y., Tang X.R., Zeng Y.J., Pan X., Wu D.H., Dong Z.Y. // Nat. Commun. 2023. V. 14. P. 1247. https://doi.org/10.1038/s41467-023-36892-4
35. Kitajima S., Tani T., Springer B.F., Campisi M., Osaki T., Haratani K., Chen M., Knelson E.H., Mahadevan N.R., Ritter J., Yoshida R., Köhler J., Ogino A., Nozawa R.S., Sundararaman S.K., Thai T.C., Homme M., Piel B., Kivlehan S., Obua B.N., Purcell C., Yajima M., Barbie T.U., Lizotte P.H., Jänne P.A., Paweletz C.P., Gokhale P.C., Barbie D.A. // Cancer Cell. 2022. V. 40. P. 1128–1144.e8. https://doi.org/10.1016/j.ccell.2022.08.015
36. He Q., Sun C., Pan Y. // Sci Rep. 2024. V. 14. P. 100. https://doi.org/10.1038/s41598-023-50703-2

GOST	Arutyunyan I., Balchir D., Fatkhudinov T., Gantsova E., Gordon K., Kananykhina E., Saburov V., Shakirova K., Soboleva A., Vishnyakova P. Effect of Mutation in the LLC1 Mouse Lewis Lung Adenocarcinoma Line on Sensitivity to Particle Radiotherapy // Russian Journal of Bioorganic Chemistry. – 2025. – V. 51. – Issue number 5 C. 953-965 . URL: https://bioorgchemras.ru/s19982860s0132342325050204-1/?version_id=120727. DOI: 10.7868/S1998286025050204
MLA	Arutyunyan, I.V, Balchir, D.V, Fatkhudinov, T.Kh, Gantsova, E.A, Gordon, K.B, Kananykhina, E.Yu, Saburov, V.O, Shakirova, K.M, Soboleva, A.G, Vishnyakova, P.A "Effect of Mutation in the LLC1 Mouse Lewis Lung Adenocarcinoma Line on Sensitivity to Particle Radiotherapy." Russian Journal of Bioorganic Chemistry. 51.5 (2025).:953-965. DOI: 10.7868/S1998286025050204
APA	Arutyunyan I., Balchir D., Fatkhudinov T., Gantsova E., Gordon K., Kananykhina E., Saburov V., Shakirova K., Soboleva A., Vishnyakova P. (2025). Effect of Mutation in the LLC1 Mouse Lewis Lung Adenocarcinoma Line on Sensitivity to Particle Radiotherapy. Russian Journal of Bioorganic Chemistry. vol. 51, no. 5, pp.953-965 DOI: 10.7868/S1998286025050204

RAS BiologyБиоорганическая химия Russian Journal of Bioorganic Chemistry

Effect of Mutation in the LLC1 Mouse Lewis Lung Adenocarcinoma Line on Sensitivity to Particle Radiotherapy

You can

References

Индексирование

RAS BiologyБиоорганическая химия Russian Journal of Bioorganic Chemistry

Effect of Mutation in the LLC1 Mouse Lewis Lung Adenocarcinoma Line on Sensitivity to Particle Radiotherapy

You can

References

Индексирование

Via social network