DETERMINATION OF METASTATIC CAPACITY IN PRIMARY LUNG CANCER CELLS: REFLECTION OF PATIENT PROFILE IN THE CLINIC USING IN VITRO METHODS

Year 2023, Volume: 6 Issue: 3, 302 - 311, 24.10.2023

Seçil Yılmaz , Medine Doğan Sarıkaya , Elif Yaşar , Burcu Şen Bağcı , Özlem Canöz , Ömer Önal

https://doi.org/10.26650/JARHS2023-1221034

Abstract

Objective: There is a scarcity of in vivo models that accurately reflect tumor growth and metastasis in cancer research. Research using cell lines with increasing passage numbers may give misleading results because the tumor loses its characteristic feature. Primary culture is the best method to represent the cellular profile of cancer patients in the laboratory environment. Therefore, we worked with patient-derived primary culture. The most common subtype of the most diagnosed lung cancer worldwide is Non-Small Cell Lung Cancer (NSCLC). Therefore, we aimed to determine patient-specific metastatic capacities by comparatively examining the migration abilities of primary cancer cells of NSCLC patients on the same platform.
Materials and methods: The migration abilities of primary cancer cells of NSCLC patients were demonstrated through wound healing assays on cisplatin and non-cisplatin groups, and measurements were made with Image J software.
Results: In the results of the wound healing assays performed on the cancer cells of five patients, it was observed that there was correlation between the wound widths, wound areas, wound closure percentages, and metastasis in the groups with and without cisplatin in Patient 3, Patient 4, and Patient 5.
Conclusion: To reflect the profile of patients visiting the clinic using patient-derived primary tumor cells, the wound healing assay can be used as a tool to demonstrate tumor behaviors, such as the patients’ responses to treatment and their metastasis-forming capacity. Detailed studies are needed in a larger population so that the physician can use in vitro tools in the decision support mechanism.

Keywords

Lung cancer, primary culture, migration, wound healing assay

Project Number

THD_2021_11187 (BAP), 215S849 (TÜBİTAK)

PRİMER AKCİĞER KANSER HÜCRELERİNDE METASTATİK KAPASİTENİN BELİRLENMESİ: KLİNİKTEKİ HASTA PROFİLİNİN İN VİTRO YÖNTEMLERLE YANSITILMASI

Abstract

Amaç: Kanser çalışmalarında tümöre dair büyüme ve metastaz durumunu birebir yansıtan in vivo modellerin azlığı söz konusudur. Hücre hatları kullanılan çalışmalarda ise pasaj sayıları ilerledikçe tümörün karakteristik özelliğini kaybetmesinden dolayı yanıltıcı sonuçlar verebilmektedir. Kanser hastalarının hücresel profilini laboratuvar ortamına taşıyabilmek için en iyi yöntem primer kültürdür. Bu nedenle çalışmamızda hasta kaynaklı primer kültür ile çalıştık. Dünya çapında en çok tanı konulan akciğer kanserinin en sık görülen alt tipi Küçük Hücreli Dışı Akciğer Kanseri (KHDAK)’dir. Bu nedenle çalışmamızda KHDAK tanısı alan hastalardan elde edilen primer kanser hücrelerinin migrasyon yeteneklerinin tek bir platformda karşılaştırmalı olarak incelenerek hastaya özgü metastatik kapasitelerinin belirlenmesi amaçlanmıştır.
Gereç ve Yöntem: KHDAK hastalarına ait primer kanser hücrelerinin migrasyon yetenekleri yara iyileştirmesi deneyiyle cisplatin içeren ve içermeyen grup üzerinde gösterilmiştir ve ölçümler Image J yazılımıyla yapılmıştır.
Bulgular: Beş hastanın kanser hücresine ait yapılan yara iyileşmesi deneyi sonuçlarında; Hasta 3, Hasta 4 ve Hasta 5’e ait cisplatin içeren ve içermeyen gruplarda yara genişlikleri, yara alanları ve yara bölgesini kapatma yüzdeleri ile metastaz arasında ilişki olduğu gözlendi.
Sonuç: Kliniğe başvuran hasta profilinin in vitro ortama en iyi şekilde yansıması için hasta kaynaklı primer tümör hücreleri kullanılarak hastaların tedaviye karşı vermiş oldukları yanıtların çeşitliliği ve metastaz oluşturma kapasiteleri gibi birçok tümör davranışının gösterilmesinde yara iyileşmesi deneyi bir araç olarak kullanılabilir. Hekimin karar destek mekanizmasında in vitro araç olarak kullanılabilmesi için daha geniş popülasyonda detaylı çalışmalara ihtiyaç duyulmaktadır.

Keywords

Akciğer Kanseri, primer kültür, migrasyon, yara iyileşmesi deneyi

Supporting Institution

Erciyes Üniversitesi BAP, TÜBİTAK

Project Number

THD_2021_11187 (BAP), 215S849 (TÜBİTAK)

Thanks

Bu çalışma kapsamında yardımlarından ötürü Nilhan Mutlu’ya, maddi desteklerinden ötürü Erciyes Üniversitesi BAP birimine ve TÜBİTAK’a teşekkür ederiz.

References

• Siegel RL, Miller KD, Fuchs HE, Jemal A. Cancer statistics, 2022. CA Cancer J Clin 2022;72(1):7-33. google scholar
• Howlader N, Forjaz G, Mooradian MJ, Meza R, Kong CY, Cronin KA, et al. The effect of advances in lung-cancer treatment on population mortality. N Engl J Med 2020;383(7):640-9. google scholar
• Cengiz H, Demirci A, Varım C, Mandel NM, Turna Z. Küçük hücreli dışı akciğer kanserinde neoadjuvan ve adjuvan tedavi sonuçları. Sakarya Tıp Dergisi 2020;10(3):450-8. google scholar
• Riihimaki M, Hemminki A, Fallah M, Thomsen H, Sundquist K, Sundquist J, et al. Metastatic sites and survival in lung cancer. Lung Cancer 2014;86(1):78-84. google scholar
• Popper HH. Progression and metastasis of lung cancer. Cancer Metastasis Rev 2016;35(1):75-91. google scholar
• Achen MG, Stacker SA. Molecular control of lymphatic metastasis. Ann N Y Acad Sci 2008;1131:225-34. google scholar
• Wu Q, Zhang B, Li B, Cao X, Chen X, Xue Q. PTBP3 promotes migration of non-small cell lung cancer through regulating E-cadherin in EMT signaling pathway. Cancer Cell Int 2020;20:172. google scholar
• Deisboeck TS, Couzin ID. Collective behavior in cancer cell populations. Bioessays 2009;31(2):190-7. google scholar
• Friedl P, Gilmour D. Collective cell migration in morphogenesis, regeneration and cancer. Nat Rev Mol Cell Biol 2009;10(7):445-57. google scholar
• Riahi R, Yang Y, Zhang DD, Wong PK. Advances in wound-healing assays for probing collective cell migration. J Lab Autom 2012;17(1):59-65. google scholar
• Bahar E, Yoon H. Modeling and predicting the cell migration properties from scratch wound healing assay on cisplatin-resistant ovarian cancer cell lines using artificial neural network. Healthcare (Basel) 2021;9(7):911. google scholar
• Kauanova S, Urazbayev A, Vorobjev I. The Frequent sampling of wound scratch assay reveals the “opportunity” window for quantitative evaluation of cell motility-impeding drugs. Front Cell Dev Biol 2021;9:640972. google scholar
• DeRose YS, Wang G, Lin YC, Bernard PS, Buys SS, Ebbert MT, et al. Tumor grafts derived from women with breast cancer authentically reflect tumor pathology, growth, metastasis and disease outcomes. Nat Med 2011;17(11):1514-20. google scholar
• Kondo J, Inoue M. Application of cancer organoid model for drug screening and personalized therapy. Cells 2019;8(5):470. google scholar
• Hait WN. Anticancer drug development: the grand challenges. Nat Rev Drug Discov 2010;9(4):253-4. google scholar
• Katt ME, Placone AL, Wong AD, Xu ZS, Searson PC. In vitro tumor models: Advantages, disadvantages, variables, and selecting the right platform. Front Bioeng Biotechnol 2016;4:12. google scholar
• Kaur G, Dufour JM. Cell lines: Valuable tools or useless artifacts. Spermatogenesis 2012;2(1):1-5. google scholar
• Fallahi P, Ferrari SM, Elia G, Ragusa F, Patrizio A, Paparo SR, et al. Primary cell cultures for the personalized therapy in aggressive thyroid cancer of follicular origin. Semin Cancer Biol 2022;79:203-16. google scholar
• Mitra A, Mishra L, Li S. Technologies for deriving primary tumor cells for use in personalized cancer therapy. Trends Biotechnol 2013;31(6):347-54. google scholar
• Grada A, Otero-Vinas M, Prieto-Castrillo F, Obagi Z, Falanga V. Research techniques made aimple: Analysis of collective cell migration using the wound healing assay. J Invest Dermatol 2017;137(2):e11-6. google scholar
• Fangjun L, Zhijia Y. Tumor suppressive roles of eugenol in human lung cancer cells. Thorac Cancer 2018;9(1):25-9. google scholar
• Schettino C, Bareschino MA, Rossi A, Maione P, Sacco PC, Colantuoni G, et al. Targeting angiogenesis for treatment of NSCLC brain metastases. Curr Cancer Drug Targets 2012;12(3):289-99. google scholar
• Spano D, Heck C, De Antonellis P, Christofori G, Zollo M. Molecular networks that regulate cancer metastasis. Semin Cancer Biol 2012;22(3):234-49. google scholar
• Fares J, Fares MY, Khachfe HH, Salhab HA, Fares Y. Molecular principles of metastasis: a hallmark of cancer revisited. Signal Transduct Target Ther 2020;5(1):28. google scholar
• Bravo-Cordero JJ, Hodgson L, Condeelis J. Directed cell invasion and migration during metastasis. Curr Opin Cell Biol 2012;24(2):277-83. google scholar
• Simpson KJ, Selfors LM, Bui J, Reynolds A, Leake D, Khvorova A, et al. Identification of genes that regulate epithelial cell migration using an siRNA screening approach. Nat Cell Biol 2008;10(9):1027-38. google scholar
• Friedl P, Wolf K. Tumour-cell invasion and migration: Diversity and escape mechanisms. Nat Rev Cancer 2003;3(5):362-74. google scholar
• Wang X, Decker CC, Zechner L et al. In vitro wound healing of tumor cells: inhibition of cell migration by selected cytotoxic alkaloids. BMC Pharmacol Toxicol 2019;20:4. google scholar
• Tonnesen MG, Feng X, Clark RA. Angiogenesis in wound healing. J Invest Derm Symp Proc 2000;5(1):40-6. google scholar
• Small JV, Geiger B, Kaverina I, Bershadsky A. How do microtubules guide migrating cells? Nat Rev Mol Cell Bio 2002;3(12):957-64. google scholar
• Ridley AJ, Schwartz MA, Burridge K, Firtel RA, Ginsberg MH, Borisy G, Parsons JT, Horwitz AR. Cell migration: integrating signals from front to back. Science. 2003;302(5651):1704-9. google scholar
• Pjuan J, Barcelo C, Moreno DF, Maiques O, Siso P, Marti RM, et al. In vitro cell migration, invasion, and adhesion assays: From Cell Imaging to Data Analysis. Front Cell Dev Biol 2019;7:107. google scholar
• Jonkman JE, Cathcart JA, Xu F, Bartolini ME, Amon JE, Stevens KM et al. An introduction to the wound healing assay using live-cell microscopy. Cell Adh Migr 2014;8(5):440-51. google scholar
• Sullivan R, Holden T, Tremberger G, Jr J, Cheung E, Branch C, et al. Fractal dimension of breast cancer cell migration in a wound healing assay. Int J Biomed Biol Engine 2008; 2(8):186-91. google scholar
• Pirker R. Adjuvant chemotherapy of non-small cell lung cancer. Tanaffos 2012;11(1):12-7. google scholar
• Han XJ, Yang ZJ, Jiang LP, Wei YF, Liao MF, Qian Y, Li Y, Huang X, Wang JB, Xin HB, Wan YY. Mitochondrial dynamics regulates hypoxia-induced migration and antineoplastic activity of cisplatin in breast cancer cells. Int J Oncol 2015;46(2):691-700. google scholar
• Manguinhas R, Fernandes AS, Costa JG, Saraiva N, Camöes SP, Gil N, et al. Impact of the APE1 redox function inhibitor E3330 in non-small cell lung cancer cells exposed to cisplatin: increased cytotoxicity and impairment of cell migration and invasion. Antioxidants 2020;9(6): 550. google scholar
• Maiuthed A, Chanvorachote P. Cisplatin at sub-toxic levels mediates integrin switch in lung cancer cells. Anticancer Res 2014;34(12):7111-7. google scholar
• Wang H, Zhang G, Zhang H, Zhang F, Zhou B, Ning F, et al. Acquisition of epithelial-mesenchymal transition phenotype and cancer stem cell-like properties in cisplatin-resistant lung cancer cells through AKT/Ş-catenin/Snail signaling pathway. Eur J Pharmacol 2014;723:156-66. google scholar