Effect of STK11 Mutation in the LLC1 Mouse Lewis Lung Adenocarcinoma Line on Sensitivity to Particle Radiotherapy

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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 Stk11 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 in vitro model of lung cancer based on the LLC1 cell line with knockout of the Stk11 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, Mki67 marker expression), apoptosis induction (annexin V-PI method) and Pten 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 Stk11 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, Mki67 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 Pten gene in mutant cells did not change significantly after irradiation. Thus, the Stk11 mutation contributes to the formation of radioresistance in tumor cells by triggering various adaptation mechanisms. The obtained in vitro model can be used for further study of radioresistance and development of new approaches to the therapy of tumors with STK11 mutation.

About the authors

E. A Gantsova

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

Email: gantsova_ea@rudn.ru
Moscow, Russia; Moscow, Russia

I. V Arutyunyan

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

Moscow, Russia; Moscow, Russia

A. G Soboleva

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

Moscow, Russia

K. M Shakirova

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

Moscow, Russia; Moscow, Russia

E. Yu Kananykhina

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

Moscow, Russia

D. V Balchir

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

Moscow, Russia

P. A Vishnyakova

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

Moscow, Russia

V. O Saburov

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

Obninsk, Russia

K. B Gordon

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 Russian Federation

Moscow, Russia; Obninsk, Russia

T. Kh Fatkhudinov

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

Moscow, Russia; Moscow, Russia

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

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