Relationship Between ISX Gene Expression and Cancer Stem Cell Markers in Hepatocellular Carcinoma Cells Exposed to Microenvironmental Changes
Öz
Aim: This study aimed to demonstrate the homeobox (ISX) protein, which is a newly identified proto-oncogene and known to be associated with hypervitaminosis A disease at the level of ISX gene expression by culturing hepatocellular carcinoma (HCC) cell line in different extracellular matrix media and to show the effect of culture media differences on cancer stem cell markers.
Material and Methods: ISX expression was 4 times higher in Matrigel culture and 2 times higher in type I collagen culture compared to the control group. Expression of the Oct-4 antibody was significantly higher in type I collagen culture medium compared to the control group (p<0.05). Nanog antibody showed a significant decrease in HepG2 cells in Matrigel culture compared to the control (p<0.05), while Ov-6 and AFP expression was highest in type I collagen, but no statistically significant difference was found (p>0.05).
Results: ISX expression was detected 4-fold higher in the Matrigel and 2-fold higher in the type I collagen culture than the control group. The Oct-4 antibody is significantly higher in the type I collagen culture (p<0.05) and the Nanog antibody showed a significant decrease in the Matrigel culture than the control group cells (p<0.05). Ov-6 and AFP antibodies are the highest type I collagen but there are no statistical significance was found (p>0.05).
Conclusion: It is thought that the results obtained from this study may help further studies on the mechanisms of liver cancer and the development of treatment options that will affect the behavior of cancer stem cells.
Anahtar Kelimeler
Hepatocellular carcinoma Vitamin A ISX gene Type 1 collagen Matrigel
Kaynakça
- 1. McGlynn, K. A., Petrick, J. L., & El-Serag, H. B. (2021). Epidemiology of Hepatocellular Carcinoma. Hepatology (Baltimore, Md.), 73 Suppl 1(Suppl 1), 4–13.
- 2. Fan, J., Chen, Y., Zhang, D., Yao, J., Zhao, Z., Jiang, Y., Li, Y., & Guo, Y. (2020). Evaluation of the diagnostic accuracy of des-gamma-carboxy prothrombin and alpha-fetoprotein alone or in combination for hepatocellular carcinoma: A systematic review and meta-analysis. Surgical oncology, 34, 245–255.
- 3. Dawson M. I. (2000). The importance of vitamin A in nutrition. Current pharmaceutical design, 6(3), 311–325.
- 4. Hsu, S. H., Wang, L. T., Lee, K. T., Chen, Y. L., Liu, K. Y., Suen, J. L., Chai, C. Y., & Wang, S. N. (2013). Proinflammatory homeobox gene, ISX, regulates tumor growth and survival in hepatocellular carcinoma. Cancer research, 73(2), 508–518.
- 5. Di Carlo, I., Mannino, M., Toro, A., Ardiri, A., Galia, A., Cappello, G., & Bertino, G. (2012). Persistent increase in alpha-fetoprotein level in a patient without underlying liver disease who underwent curative resection of hepatocellular carcinoma. A case report and review of the literature. World journal of surgical oncology, 10, 79.
- 6. Galle, P. R., Foerster, F., Kudo, M., Chan, S. L., Llovet, J. M., Qin, S., Schelman, W. R., Chintharlapalli, S., Abada, P. B., Sherman, M., & Zhu, A. X. (2019). Biology and significance of alpha-fetoprotein in hepatocellular carcinoma. Liver international: official journal of the International Association for the Study of the Liver, 39(12), 2214–2229.
- 7. Zhang JL, Gong LQ, Yan Q, Zhou NN, Lee VHF, Guan XY. (2019) Advances in surface markers of liver cancer stem cell. Hepatoma Res 5(27),1-18.
- 8. Yang, W., Wang, C., Lin, Y., Liu, Q., Yu, L. X., Tang, L., Yan, H. X., Fu, J., Chen, Y., Zhang, H. L., Tang, L., Zheng, L. Y., He, Y. Q., Li, Y. Q., Wu, F. Q., Zou, S. S., Li, Z., Wu, M. C., Feng, G. S., & Wang, H. Y. (2012). OV6⁺ tumor-initiating cells contribute to tumor progression and invasion in human hepatocellular carcinoma. Journal of hepatology, 57(3), 613–620.
- 9. Mahalaxmi, I., Devi, S. M., Kaavya, J., Arul, N., Balachandar, V., & Santhy, K. S. (2019). New insight into NANOG: A novel therapeutic target for ovarian cancer (OC). European journal of pharmacology, 852, 51–57.
- 10. Mohiuddin, I. S., Wei, S. J., & Kang, M. H. (2020). Role of OCT4 in cancer stem-like cells and chemotherapy resistance. Biochimica et biophysica acta. Molecular basis of disease, 1866(4), 165432.
- 11. Nissen, N. I., Karsdal, M., & Willumsen, N. (2019). Collagens and Cancer associated fibroblasts in the reactive stroma and its relation to Cancer biology. Journal of experimental & clinical cancer research: CR, 38(1), 115.
- 12. Hielscher, A., McGuire, T. , Weisenburger, D. and Sharp, J. (2013).Matrigel modulates a stem cell phenotype and promotes tumor formation in a mantle cell lymphoma cell line. Stem Cell Discovery, 3(3),167-179.
- 13. Liu, K. Y., Wang, L. T., Hsu, S. H., & Wang, S. N. (2019). Homeobox Genes and Hepatocellular Carcinoma. Cancers, 11(5), 621.
- 14. Zheng, X., Liu, W., Xiang, J., Liu, P., Ke, M., Wang, B., Wu, R., & Lv, Y. (2017). Collagen I promotes hepatocellular carcinoma cell proliferation by regulating integrin β1/FAK signaling pathway in nonalcoholic fatty liver. Oncotarget, 8(56), 95586–95595.
- 15. Zhang JL, Gong LQ, Yan Q, Zhou NN, Lee VHF, Guan XY. (2019) Advances in surface markers of liver cancer stem cell. Hepatoma Res 5(27),1-18.
- 16. Liang, C., Zhang, K., Ge, H., Li, W., Li, G., & Wu, J. (2018). Prognostic and clinicopathological value of Nanog in hepatocellular carcinoma: A meta-analysis. Clinica chimica acta; international journal of clinical chemistry, 477, 24–31.
- 17. Vasefifar, P., Najafi, S., Motafakkerazad, R., Amini, M., Safaei, S., Najafzadeh, B., Alemohammad, H., Jafarlou, M., & Baradaran, B. (2023). Targeting Nanog expression increased Cisplatin chemosensitivity and inhibited cell migration in Gastric cancer cells. Experimental cell research, 429(2), 113681.
- 18. Khosravi, N., Shahgoli, V. K., Amini, M., Safaei, S., Mokhtarzadeh, A., Mansoori, B., Derakhshani, A., Baghbanzadeh, A., & Baradaran, B. (2021). Suppression of Nanog inhibited cell migration and increased the sensitivity of colorectal cancer cells to 5-fluorouracil. European journal of pharmacology, 894, 173871.
- 19. Kosaka, T., Mikami, S., Yoshimine, S., Miyazaki, Y., Daimon, T., Kikuchi, E., Miyajima, A., & Oya, M. (2016). The prognostic significance of OCT4 expression in patients with prostate cancer. Human pathology, 51, 1–8.
- 20. Dong, Z., Zeng, Q., Luo, H., Zou, J., Cao, C., Liang, J., Wu, D., & Liu, L. (2012). Increased expression of OCT4 is associated with low differentiation and tumor recurrence in human hepatocellular carcinoma. Pathology, research and practice, 208(9), 527-533.
Mikroçevre değişimlerine maruz kalan hepatosellüler karsinom hücrelerinde ISX gen ekspresyonu ile kanser kök hücre belirteçleri arasındaki ilişki
Öz
Amaç: Bu çalışmada yeni tanımlanan bir proto-onkogen olan ve vücutta çok fazla A vitamini birikmesiyle ortaya çıkan hipervitaminoz A hastalığıyla ilişkili olduğu bilinen homeobox (ISX) proteininin, Hepatoselüler karsinom (HCC) hücre hattının farklı ekstrasellüler matriks ortamlarda kültüre edilmesiyle, ISX gen ekspresyonu düzeyinde gösterilmesi, ayrıca kültür ortam faklılıklarının kanser kök hücre belirteçleri üzerindeki etkisinin değerlendirmesi amaçlanmıştır.
Materyal ve Metotlar: Karaciğer karsinom hücre hattı olan HepG2 hücreler tip I kollajen ve matrigel kültür ortamlardakültüre edilerek, ISX geninin ekspresyonu real time PCR kullanılarak, alfa-fetoprotein (AFP), kök hücre belirteçleri olan OV-6 ve Nanog proteinlerinin hücresel miktarları ve lokalizasyonlarındaki değişiklikler konfokal mikroskop ile araştırılmıştır.
Bulgular: ISX gen ekspresyonu, kontrol grubuna göre Matrigel kültür ortamda 4 kat, tip I kolajen kültürde ise 2 kat daha fazla tespit edilmiştir. Oct-4 antikorunun ekspresyonu tip I kollajen kültür ortamında kontrol grubuna göre istatistiksel olarak anlamlı derecede yüksektir (p<0,05). Nanog antikoru, kontrol grubuna göre Matrigel ortamda HepG2 hücrelerinde anlamlı bir azalma gösterirken (p<0,05), Ov-6 ve AFP ekspresyonu en yüksek tip I kollajen ortamda görülmüş ancak istatistiksel olarak anlamlı bir fark bulunmamıştır (p>0,05).
Sonuç: Bu çalışmadan elde edilen sonuçların karaciğer kanseri mekanizmaları ve kanser kök hücrelerinin davranışını etkileyecek tedavi seçeneklerinin geliştirilmesi konusunda yapılacak ileri çalışmalara yardımcı olabileceği düşünülmektedir.
Anahtar Kelimeler
Hepatoselüler karsinom A vitamini ISX geni Tip1 kollajen Matrigel
Etik Beyan
Hücre Kültürü Çalışması
Destekleyen Kurum
TÜBİTAK 2209A Lisans Öğrencileri Destekleme Programı
Kaynakça
- 1. McGlynn, K. A., Petrick, J. L., & El-Serag, H. B. (2021). Epidemiology of Hepatocellular Carcinoma. Hepatology (Baltimore, Md.), 73 Suppl 1(Suppl 1), 4–13.
- 2. Fan, J., Chen, Y., Zhang, D., Yao, J., Zhao, Z., Jiang, Y., Li, Y., & Guo, Y. (2020). Evaluation of the diagnostic accuracy of des-gamma-carboxy prothrombin and alpha-fetoprotein alone or in combination for hepatocellular carcinoma: A systematic review and meta-analysis. Surgical oncology, 34, 245–255.
- 3. Dawson M. I. (2000). The importance of vitamin A in nutrition. Current pharmaceutical design, 6(3), 311–325.
- 4. Hsu, S. H., Wang, L. T., Lee, K. T., Chen, Y. L., Liu, K. Y., Suen, J. L., Chai, C. Y., & Wang, S. N. (2013). Proinflammatory homeobox gene, ISX, regulates tumor growth and survival in hepatocellular carcinoma. Cancer research, 73(2), 508–518.
- 5. Di Carlo, I., Mannino, M., Toro, A., Ardiri, A., Galia, A., Cappello, G., & Bertino, G. (2012). Persistent increase in alpha-fetoprotein level in a patient without underlying liver disease who underwent curative resection of hepatocellular carcinoma. A case report and review of the literature. World journal of surgical oncology, 10, 79.
- 6. Galle, P. R., Foerster, F., Kudo, M., Chan, S. L., Llovet, J. M., Qin, S., Schelman, W. R., Chintharlapalli, S., Abada, P. B., Sherman, M., & Zhu, A. X. (2019). Biology and significance of alpha-fetoprotein in hepatocellular carcinoma. Liver international: official journal of the International Association for the Study of the Liver, 39(12), 2214–2229.
- 7. Zhang JL, Gong LQ, Yan Q, Zhou NN, Lee VHF, Guan XY. (2019) Advances in surface markers of liver cancer stem cell. Hepatoma Res 5(27),1-18.
- 8. Yang, W., Wang, C., Lin, Y., Liu, Q., Yu, L. X., Tang, L., Yan, H. X., Fu, J., Chen, Y., Zhang, H. L., Tang, L., Zheng, L. Y., He, Y. Q., Li, Y. Q., Wu, F. Q., Zou, S. S., Li, Z., Wu, M. C., Feng, G. S., & Wang, H. Y. (2012). OV6⁺ tumor-initiating cells contribute to tumor progression and invasion in human hepatocellular carcinoma. Journal of hepatology, 57(3), 613–620.
- 9. Mahalaxmi, I., Devi, S. M., Kaavya, J., Arul, N., Balachandar, V., & Santhy, K. S. (2019). New insight into NANOG: A novel therapeutic target for ovarian cancer (OC). European journal of pharmacology, 852, 51–57.
- 10. Mohiuddin, I. S., Wei, S. J., & Kang, M. H. (2020). Role of OCT4 in cancer stem-like cells and chemotherapy resistance. Biochimica et biophysica acta. Molecular basis of disease, 1866(4), 165432.
- 11. Nissen, N. I., Karsdal, M., & Willumsen, N. (2019). Collagens and Cancer associated fibroblasts in the reactive stroma and its relation to Cancer biology. Journal of experimental & clinical cancer research: CR, 38(1), 115.
- 12. Hielscher, A., McGuire, T. , Weisenburger, D. and Sharp, J. (2013).Matrigel modulates a stem cell phenotype and promotes tumor formation in a mantle cell lymphoma cell line. Stem Cell Discovery, 3(3),167-179.
- 13. Liu, K. Y., Wang, L. T., Hsu, S. H., & Wang, S. N. (2019). Homeobox Genes and Hepatocellular Carcinoma. Cancers, 11(5), 621.
- 14. Zheng, X., Liu, W., Xiang, J., Liu, P., Ke, M., Wang, B., Wu, R., & Lv, Y. (2017). Collagen I promotes hepatocellular carcinoma cell proliferation by regulating integrin β1/FAK signaling pathway in nonalcoholic fatty liver. Oncotarget, 8(56), 95586–95595.
- 15. Zhang JL, Gong LQ, Yan Q, Zhou NN, Lee VHF, Guan XY. (2019) Advances in surface markers of liver cancer stem cell. Hepatoma Res 5(27),1-18.
- 16. Liang, C., Zhang, K., Ge, H., Li, W., Li, G., & Wu, J. (2018). Prognostic and clinicopathological value of Nanog in hepatocellular carcinoma: A meta-analysis. Clinica chimica acta; international journal of clinical chemistry, 477, 24–31.
- 17. Vasefifar, P., Najafi, S., Motafakkerazad, R., Amini, M., Safaei, S., Najafzadeh, B., Alemohammad, H., Jafarlou, M., & Baradaran, B. (2023). Targeting Nanog expression increased Cisplatin chemosensitivity and inhibited cell migration in Gastric cancer cells. Experimental cell research, 429(2), 113681.
- 18. Khosravi, N., Shahgoli, V. K., Amini, M., Safaei, S., Mokhtarzadeh, A., Mansoori, B., Derakhshani, A., Baghbanzadeh, A., & Baradaran, B. (2021). Suppression of Nanog inhibited cell migration and increased the sensitivity of colorectal cancer cells to 5-fluorouracil. European journal of pharmacology, 894, 173871.
- 19. Kosaka, T., Mikami, S., Yoshimine, S., Miyazaki, Y., Daimon, T., Kikuchi, E., Miyajima, A., & Oya, M. (2016). The prognostic significance of OCT4 expression in patients with prostate cancer. Human pathology, 51, 1–8.
- 20. Dong, Z., Zeng, Q., Luo, H., Zou, J., Cao, C., Liang, J., Wu, D., & Liu, L. (2012). Increased expression of OCT4 is associated with low differentiation and tumor recurrence in human hepatocellular carcinoma. Pathology, research and practice, 208(9), 527-533.
Ayrıntılar
| Birincil Dil | İngilizce |
|---|---|
| Konular | Klinik Tıp Bilimleri (Diğer) |
| Bölüm | Araştırma Makalesi |
| Yazarlar | |
| Yayımlanma Tarihi | 29 Nisan 2025 |
| Gönderilme Tarihi | 3 Mart 2025 |
| Kabul Tarihi | 28 Nisan 2025 |
| Yayımlandığı Sayı | Yıl 2025 Cilt: 17 Sayı: 1 |
