Akciğer kanseri hastalarında dolaşımdaki tümör hücrelerini nasıl tespit ederiz? akım sitometrisi ile yapılan kısa bir çalışma
Öz
Dolaşımdaki tümör hücreleri (CTC’ler)
karsinomların metastatik yayılımında önemli role sahiptir. Bu nedenle, son
yıllarda birçok kanser türünde hastalığın seyri ve tedavi etkinliğinin
anlaşılmasında yardımcı olabileceği düşünülen CTC’ler üzerine yoğunlaşılmıştır.
Teknik yaklaşımlardaki gelişmeler ile özellikle akciğer kanseri gibi dokularına
ulaşılması zor olan tümörlerde sıvı biyopsi olarak tüm kandan CTC tespitinin
değeri her geçen gün artmaktadır. Tanı anında sıklıkla metastaz yaptıkları
teşhis edilen akciğer kanseri, hem kadınlarda hem de erkeklerde kanser ölüm
nedenlerinin birincil sebebidir. Çalışmamızın amacı akım sitometrisi ile
akciğer kanserli hastaların periferik kan örneklerinde (7.5 mL) CTC tespitidir.
Zenginleştirme ve saptama adımlarından oluşan modifiye ettiğimiz yöntemimiz ile
CTC sayısının tespit edilmesi için 9 akciğer kanserli birey ve 9 sağlıklı birey
çalışmaya dahil edilmiştir. Uyguladığımız metotta zenginleştirme basamağı için
fikol yoğunluk gradiyent ayrımı ve immünomanyetik ayırma tekniği (CD45 negatif
seçilim) gerçekleştirilmiştir. Sonrasında, zenginleştirilmiş hücreler arasında
CTC’leri tespit etmek için, anti-epitelyal hücre adhezyon molekülünün ve
sitokeratinlerin ekspresyonuna dayanan çok parametreli akım sitometrisi ile
analiz yapılmıştır. Çalışmamız sonucunda akciğer kanserli hastaların tümünde
CTC gözlenirken, sağlıklı bireylerde gözlenmemiştir (Z=3.823; p<0.001).
Böylece, modifiye ettiğimiz metodun akciğer kanserinde CTC tespitinde
kullanılabilirliği gösterilmiştir.
Anahtar Kelimeler
Dolaşımdaki tümör hücreleri akım sitometresi akciğer kanseri
Kaynakça
- 1. The Globocan Project. Cancer Fact Sheets: Population Fact Sheets. http://globocan.iarc.fr/Pages/fact_sheets_population.aspx. Accesed June 06, 2016.
- 2. Turkey Cancer Statistics. http://kanser.gov.tr/Dosya/ca_istatistik/ANA_rapor_2013v01_2.pdf. Accesed June 06, 2016.
- 3. Truini A, Alama A, Dal Bello MG, et al. Clinical applications of circulating tumor cells in lung cancer patients by cellsearch system. Front Oncol. 2014;4:242.
- 4. The Globocan Project. Lung Cancer Estimated Incidence, Mortality and Prevalence Worldwide. http://globocan.iarc.fr/old/FactSheets/cancers/lung-new.asp. Accesed June 24, 2016.
- 5. Spiro SG, Porter JC. Lung cancer-where are we today? Current advances in staging and nonsurgical treatment. Am J Respir Crit Care Med. 2002;166(9):1166-1196.
- 6. Wong BS, Hsiao YC, Lin TW, et al. The in vitro and in vivo apoptotic effects of Mahonia oiwakensis on human lung cancer cells. Chem Biol Interact. 2009;180(2):165-174.
- 7. Beckles MA, Spiro SG, Colice GL, et al. Initial evaluation of the patient with lung cancer: symptoms, signs, laboratory tests, and paraneoplastic syndromes. Chest. 2003;123:97-104.
- 8. Chambers AF. The metastatic process: basic research and clinical implications. Oncol Res. 1999;11(4):161-168.
- 9. Gao W, Yuan H, Jing F, et al. Analysis of circulating tumor cells from lung cancer patients with multiple biomarkers using high-performance size-based microfluidic chip. Oncotarget. 2017;8(8):12917-12928.
- 10. Rolfo C, Castiglia M, Hong D, et al. Liquid biopsies in lung cancer: the new ambrosia of researchers. Biochim Biophys Acta. 2014;1846(2):539-546.
- 11. Zhang Z, Ramnath N, Nagrath S. Current status of CTCs as liquid biopsy in lung cancer and future directions. Front Oncol. 2015;5:209.
- 12. Hong B, Zu Y. Detecting circulating tumor cells: current challenges and new trends. Theranostics. 2013;3(6):377-394.
- 13. Lowes LE, Allan AL. Recent advances in the molecular characterization of circulating tumor cells. Cancers (Basel). 2014;6(1):595-624.
- 14. Simsek E, Guler OO, Carhan A, et al. The determination of circulating tumor cells in peripheral blood by flow cytometry. Niche. 2014;3. doi: 10.5152/niche.2015.246.
- 15. Wang L, Wu C, Qiao L, et al. Clinical Significance of Folate Receptor-positive Circulating Tumor Cells Detected by Ligand-targeted Polymerase Chain Reaction in Lung Cancer. J Cancer. 2017;8(1):104-110.
- 16. Joosse SA, Gorges TM, Pantel K. Biology, detection, and clinical implications of circulating tumor cells. EMBO Mol Med. 2015;7(1):1-11.
- 17. Welinder C, Jansson B, Lindell G, et al. Cytokeratin 20 improves the detection of circulating tumor cells in patients with colorectal cancer. Cancer Lett. 2015;358(1):43-46.
- 18. Bevilacqua S, Gallo M, Franco R, et al. A “live” biopsy in a small-cell lung cancer patient by detection of circulating tumor cells. Lung Cancer. 2009;65(1):123-125.
- 19. Tanaka F, Yoneda K, Kondo N, et al. Circulating tumor cell as a diagnostic marker in primary lung cancer. Clin Cancer Res. 2009;15(22):6980-6986.
- 20. Krebs MG, Sloane R, Priest L, et al. Evaluation and prognostic significance of circulating tumor cells in patients with non–small-cell lung cancer. J Clin Oncol. 2011;29(12):1556-1563.
- 21. Krebs MG, Hou JM, Sloane R, et al. Analysis of circulating tumor cells in patients with non-small cell lung cancer using epithelial marker-dependent and-independent approaches. J Thorac Oncol. 2012;7(2):306-315.
- 22. Alunni-Fabbroni M, Sandri MT. Circulating tumour cells in clinical practice: Methods of detection and possible characterization. Methods. 2010;50(4):289-297.
- 23. Allard WJ, Matera J, Miller MC, et al. Tumor cells circulate in the peripheral blood of all major carcinomas but not in healthy subjects or patients with nonmalignant diseases. Clin Cancer Res. 2004;10(20):6897-6904.
- 24. Bøyum A, Løvhaug D, Tresland L, et al. Separation of leucocytes: improved cell purity by fine adjustments of gradient medium density and osmolality. Scand J Immunol. 1991;34(6):697-712.
- 25. Liu Z, Fusi A, Klopocki E, et al. Negative enrichment by immunomagnetic nanobeads for unbiased characterization of circulating tumor cells from peripheral blood of cancer patients. J Transl Med. 2011;9:70.
- 26. Went P, Vasei M, Bubendorf L, et al. Frequent high-level expression of the immunotherapeutic target Ep-CAM in colon, stomach, prostate and lung cancers. Br J Cancer. 2006;94(1):128-135.
- 27. Domanski D, Perzanowska A, Kistowski M, et al. A multiplexed cytokeratin analysis using targeted mass spectrometry reveals specific profiles in cancer-related pleural effusions. Neoplasia. 2016;18(7):399-412.
- 28. Huang HB, Ge MJ. The effects of different surgical approaches on the perioperative level of circulating tumor cells in patients with non-small cell lung cancer. Thorac Cardiovasc Surg. 2016;64(6):515-519.
- 29. Ren C, Han C, Wang D, et al. Detection of circulating tumor cells: Clinical relevance of a novel metastatic tumor marker. Exp Ther Med. 2011;2(3):385-391.
How could we detect circulating tumor cells in lung cancer patients? A brief study by flow cytometry
Öz
Circulating tumor cells (CTCs) play a
crucial role in the metastatic spread of carcinoma. Therefore, CTC has been
interest of a subject in the past few decades in terms of prognosis and
response to the therapy in several cancer diseases. Recent improvements in
technical approaches maintain to identify CTCs from whole blood have
demonstrated the potential value of CTC detection as a liquid biopsy especially
in those tumors where tissue accessibility is often challenging as in lung
cancer. Lung cancer is the most common cause of death from cancer worldwide in
both men and women which is commonly metastasize before it is diagnosed. The
aim of this study is to enumerate of CTCs in peripheral blood sample (7.5 mL)
of lung cancer patients by flow cytometry. Our modified method which consists of
enrichment and detection steps get involved in 9 patients with lung cancer and
9 healthy volunteers. We performed a density-based ficoll gradient
centrifugation and a immunomagnetic separation technique (CD45 negative selection)
for the enrichment step. Next,
multi-parameter flow cytometry based on the expression of anti-epithelial cell
adhesion molecule and cytokeratins was used to detect circulating tumor cells
among enriched cells. According to our results,
circulating tumor cells were not detected on healthy volunteers but
circulating tumor cells were found in all of patients with lung cancer
(Z=3.823; p<0.001). We demonstrate that circulating tumor cells were
detectable in peripheral blood sample of lung cancer patients by our modified
method.
Anahtar Kelimeler
Kaynakça
- 1. The Globocan Project. Cancer Fact Sheets: Population Fact Sheets. http://globocan.iarc.fr/Pages/fact_sheets_population.aspx. Accesed June 06, 2016.
- 2. Turkey Cancer Statistics. http://kanser.gov.tr/Dosya/ca_istatistik/ANA_rapor_2013v01_2.pdf. Accesed June 06, 2016.
- 3. Truini A, Alama A, Dal Bello MG, et al. Clinical applications of circulating tumor cells in lung cancer patients by cellsearch system. Front Oncol. 2014;4:242.
- 4. The Globocan Project. Lung Cancer Estimated Incidence, Mortality and Prevalence Worldwide. http://globocan.iarc.fr/old/FactSheets/cancers/lung-new.asp. Accesed June 24, 2016.
- 5. Spiro SG, Porter JC. Lung cancer-where are we today? Current advances in staging and nonsurgical treatment. Am J Respir Crit Care Med. 2002;166(9):1166-1196.
- 6. Wong BS, Hsiao YC, Lin TW, et al. The in vitro and in vivo apoptotic effects of Mahonia oiwakensis on human lung cancer cells. Chem Biol Interact. 2009;180(2):165-174.
- 7. Beckles MA, Spiro SG, Colice GL, et al. Initial evaluation of the patient with lung cancer: symptoms, signs, laboratory tests, and paraneoplastic syndromes. Chest. 2003;123:97-104.
- 8. Chambers AF. The metastatic process: basic research and clinical implications. Oncol Res. 1999;11(4):161-168.
- 9. Gao W, Yuan H, Jing F, et al. Analysis of circulating tumor cells from lung cancer patients with multiple biomarkers using high-performance size-based microfluidic chip. Oncotarget. 2017;8(8):12917-12928.
- 10. Rolfo C, Castiglia M, Hong D, et al. Liquid biopsies in lung cancer: the new ambrosia of researchers. Biochim Biophys Acta. 2014;1846(2):539-546.
- 11. Zhang Z, Ramnath N, Nagrath S. Current status of CTCs as liquid biopsy in lung cancer and future directions. Front Oncol. 2015;5:209.
- 12. Hong B, Zu Y. Detecting circulating tumor cells: current challenges and new trends. Theranostics. 2013;3(6):377-394.
- 13. Lowes LE, Allan AL. Recent advances in the molecular characterization of circulating tumor cells. Cancers (Basel). 2014;6(1):595-624.
- 14. Simsek E, Guler OO, Carhan A, et al. The determination of circulating tumor cells in peripheral blood by flow cytometry. Niche. 2014;3. doi: 10.5152/niche.2015.246.
- 15. Wang L, Wu C, Qiao L, et al. Clinical Significance of Folate Receptor-positive Circulating Tumor Cells Detected by Ligand-targeted Polymerase Chain Reaction in Lung Cancer. J Cancer. 2017;8(1):104-110.
- 16. Joosse SA, Gorges TM, Pantel K. Biology, detection, and clinical implications of circulating tumor cells. EMBO Mol Med. 2015;7(1):1-11.
- 17. Welinder C, Jansson B, Lindell G, et al. Cytokeratin 20 improves the detection of circulating tumor cells in patients with colorectal cancer. Cancer Lett. 2015;358(1):43-46.
- 18. Bevilacqua S, Gallo M, Franco R, et al. A “live” biopsy in a small-cell lung cancer patient by detection of circulating tumor cells. Lung Cancer. 2009;65(1):123-125.
- 19. Tanaka F, Yoneda K, Kondo N, et al. Circulating tumor cell as a diagnostic marker in primary lung cancer. Clin Cancer Res. 2009;15(22):6980-6986.
- 20. Krebs MG, Sloane R, Priest L, et al. Evaluation and prognostic significance of circulating tumor cells in patients with non–small-cell lung cancer. J Clin Oncol. 2011;29(12):1556-1563.
- 21. Krebs MG, Hou JM, Sloane R, et al. Analysis of circulating tumor cells in patients with non-small cell lung cancer using epithelial marker-dependent and-independent approaches. J Thorac Oncol. 2012;7(2):306-315.
- 22. Alunni-Fabbroni M, Sandri MT. Circulating tumour cells in clinical practice: Methods of detection and possible characterization. Methods. 2010;50(4):289-297.
- 23. Allard WJ, Matera J, Miller MC, et al. Tumor cells circulate in the peripheral blood of all major carcinomas but not in healthy subjects or patients with nonmalignant diseases. Clin Cancer Res. 2004;10(20):6897-6904.
- 24. Bøyum A, Løvhaug D, Tresland L, et al. Separation of leucocytes: improved cell purity by fine adjustments of gradient medium density and osmolality. Scand J Immunol. 1991;34(6):697-712.
- 25. Liu Z, Fusi A, Klopocki E, et al. Negative enrichment by immunomagnetic nanobeads for unbiased characterization of circulating tumor cells from peripheral blood of cancer patients. J Transl Med. 2011;9:70.
- 26. Went P, Vasei M, Bubendorf L, et al. Frequent high-level expression of the immunotherapeutic target Ep-CAM in colon, stomach, prostate and lung cancers. Br J Cancer. 2006;94(1):128-135.
- 27. Domanski D, Perzanowska A, Kistowski M, et al. A multiplexed cytokeratin analysis using targeted mass spectrometry reveals specific profiles in cancer-related pleural effusions. Neoplasia. 2016;18(7):399-412.
- 28. Huang HB, Ge MJ. The effects of different surgical approaches on the perioperative level of circulating tumor cells in patients with non-small cell lung cancer. Thorac Cardiovasc Surg. 2016;64(6):515-519.
- 29. Ren C, Han C, Wang D, et al. Detection of circulating tumor cells: Clinical relevance of a novel metastatic tumor marker. Exp Ther Med. 2011;2(3):385-391.
Ayrıntılar
| Birincil Dil | İngilizce |
|---|---|
| Konular | Sağlık Kurumları Yönetimi |
| Bölüm | Araştırma Makalesi |
| Yazarlar | |
| Yayımlanma Tarihi | 29 Aralık 2018 |
| Gönderilme Tarihi | 28 Mart 2018 |
| Kabul Tarihi | 26 Nisan 2018 |
| Yayımlandığı Sayı | Yıl 2018 Cilt: 3 Sayı: 4 |
