Araştırma Makalesi
BibTex RIS Kaynak Göster

PATULİNİN KARACİĞER VE AKCİĞER KANSERİ HÜCRE HATLARI ÜZERİNDEKİ İN VİTRO ANTİTÜMÖR AKTİVİTESİ

Yıl 2022, Cilt: 46 Sayı: 2, 393 - 404, 29.05.2022
https://doi.org/10.33483/jfpau.1078080

Öz

Amaç: Patulin, başta Aspergillus ve Penicilium olmak üzere bazı küf mantarları tarafından üretilen bir mikotoksin olup, hayvanlarda ve insanlarda mikotoksikozdan sorumludur. Patulinin antitümör aktivitesi hakkında hala çok ayrıntılı veriler bulunmamakla birlikte bazı raporlar hücresel apoptozu ve toksisiteyi indüklediğini göstermektedir. Bizim bu çalışmadaki amacımız patulinin insan karaciğer ve akciğer kanseri hücreleri üzerindeki antiproliferatif, antimigrasyon etkilerini ve apoptotik yolaklardaki gen ifadeleri üzerine etkisini belirlemektir.
Gereç ve Yöntem: Patulinin karaciğer ve akciğer kanserleri tedavisindeki etkinliğini belirlemek için HEP3B ve A549 hücre hatları kullanıldı. Hücre hatları, artan konsantrasyonlarda patuline (1, 2.5, 5, 10, 25, 50 ve 100 µM) maruz bırakıldı, ardından MTS testi, yara iyileşme testi ve RT-PCR deneyleri ile, hücre canlılığı, hücre göçü ve apoptoz belirlendi.
Sonuç ve Tartışma: Patulin her iki kanser hücre hattında da doza bağlı olarak hücre canlılığında ve hücre göçünde önemli bir azalmaya neden oldu Ayrıca her iki hücre hattında da apoptozun indüksiyonunu teşvik etti. Sonuçlarımız patulinin insan karaciğer ve akciğer kanser hücrelerinde tümör büyümesini in vitro olarak önemli ölçüde azaltabileceğini gösterdi.

Destekleyen Kurum

İNÖNÜ ÜNİVERSİTESİ BİLİMSEL ARAŞTIRMA PROJE BİRİMİ (İUBAP)

Proje Numarası

TSG-2020-2087

Teşekkür

Bu çalışma TSG-2020-2087 numaralı İUBAP (İnönü Üniversitesi Bilimsel Araştırma Proje Birimi) tarafından desteklenmiştir.

Kaynakça

  • Referans1. Ismaiel, A.A., Papenbrock, J. (2017). Effect of patulin from penicillium vulpinum on the activity of glutathione-S-transferase and selected antioxidative enzymes in Maize. International Journal of Environmental Research and Public Health, 14(7), 825. [CrossRef]
  • Referans2. Puel, O., Galtier, P., Oswald, I.P. (2010). Biosynthesis and toxicological effects of patulin. Toxins (Basel), 2(4), 613-631. [CrossRef]
  • Referans3. Liu, B.H., Yu, F.Y., Wu, T.S., Li, S.Y., Su, M.C., Wang, M.C., Shih, S.M. (2003). Evaluation of genotoxic risk and oxidative DNA damage in mammalian cells exposed to mycotoxins, patulin and citrinin. Toxicology and Applied Pharmacology, 191(3), 255-63. [CrossRef]
  • Referans4. IARC. Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to Humans. IARC; Lyon, France: 1986. Some Naturally Occurring and Synthetic Food Components, Furocoumarins and Ultraviolet Radiation; pp. 83-98.
  • Referans5. Abastabar, M., Akbari, A., Akhtari, J., Hedayati, M. T., Shokohi, T., Mehrad-Majd, H., Ghalehnoei, H., Ghasemi, S. (2017). In vitro antitumor activity of patulin on cervical and colorectal cancer cell lines. Current Medical Mycology, 3(1), 25-29.[CrossRef]
  • Referans6. Boussabbeh, M., Ben Salem, I., Prola, A., Guilbert, A., Bacha, H., Abid-Essefi, S., Lemaire, C. (2015). Patulin induces apoptosis through ROS-mediated endoplasmic reticulum stress pathway. Toxicological Sciences, 144(2), 328-337.[CrossRef]
  • Referans7. Pal, S., Singh, N., & Ansari, K. M. (2017). Toxicological effects of patulin mycotoxin on the mammalian system: an overview. Toxicology Research, 6(6), 764–771. [CrossRef]
  • Referans8. Liu, J., Liu, Q., Han, J., Feng, J., Guo, T., Li, Z., Min, F., Jin, R., Peng, X. (2021). N-Acetylcysteine inhibits patulin-induced apoptosis by affecting ROS-mediated oxidative damage pathway. Toxins, 13(9), 595. [CrossRef]
  • Referans9. Barltrop, J.A., Owen, T.C., Cory, A.H., Cory, J.G. (1991). 5-(3- carboxymethoxyphenyl)-2-(4,5-dimethylthiazolyl)-3- (4-sulfophenyl) tetrazolium, inner salt (MTS) and related analogs of 3-(4,5-dimethylthiazolyl)-2,5- diphenyltetrazolium bromide (MTT) reducing to purple water-soluble formazans as cell-viability indicators. Bioorganic & Medicinal Chemistry Letters, 1(11), 611-614. [CrossRef]
  • Referans10. Liang, C.C., Park, A., Guan, J.L. (2007). In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro. Nature Protocols, 2(2), 329-333. [CrossRef]
  • Referans11. Demirci, S., Doğan, A., Türkmen, N. B., Telci, D., Rizvanov, A. A., Şahin, F. (2017). Schiff base-Poloxamer P85 combination demonstrates chemotherapeutic effect on prostate cancer cells in vitro. Biomedicine & Pharmacotherapy, 86, 492-501. [CrossRef]
  • Referans12. Andersen, B., Smedsgaard, J., Frisvad, J.C. (2004). Penicillium expansum: Consistent production of patulin, chaetoglobosins, and other secondary metabolites in culture and their natural occurrence in fruit products. Journal of Agricultural and Food Chemistry, 52(8), 2421-2428. [CrossRef]
  • Referans13. Kwon, O., Soung, N. K., Thimmegowda, N. R., Jeong, S. J., Jang, J. H., Moon, D. O., Chung, J. K., Lee, K. S., Kwon, Y. T., Erikson, R. L., Ahn, J. S., Kim, B. Y. (2012). Patulin induces colorectal cancer cells apoptosis through EGR-1 dependent ATF3 up-regulation. Cellular Signalling, 24(4), 943-950. [CrossRef]
  • Referans14. Liu, B. H., Wu, T. S., Yu, F. Y., Wang, C. H. (2006). Mycotoxin patulin activates the p38 kinase and JNK signaling pathways in human embryonic kidney cells. Toxicological Sciences, 89(2), 423-430. [CrossRef]
  • Referans15. Guo, X., Dong, Y., Yin, S., Zhao, C., Huo, Y., Fan, L. (2013). Patulin induces pro-survival functions via autophagy inhibition and p62 accumulation. Cell Death Disease, 4(10), e822. [CrossRef]
  • Referans16. Ferrer, E., Juan-García, A., Font, G., Ruiz, M. J. (2009). Reactive oxygen species induced by beauvericin, patulin and zearalenone in CHO-K1 cells. Toxicology In Vitro, 23(8), 1504-1509.[CrossRef]
  • Referans17. Alves, I., Oliveira, N., Laires, A., Rodrigues, A., Rueff, J. (2000). Induction of micronuclei and chromosomal aberrations by the mycotoxin patulin in mammalian cells: role of ascorbic acid as a modulator of patulin clastogenicity. Mutagenesis, 15(3), 229-34. [CrossRef]
  • Referans18. Burghardt, R. C., Barhoumi, R., Lewis, E. H., Bailey, R. H., Pyle, K. A., Clement, B. A., Phillips, T. D. (1992). Patulin-induced cellular toxicity: a vital fluorescence study. Toxicology and Applied Pharmacology, 112(2), 235-244. [CrossRef]
  • Referans19. Turkmen, N. B., Yuce, H., Ozek, D. A., Aslan, S., Yasar, S., Unuvar, S. (2021). Dose dependent cytotoxic activity of patulin on neuroblastoma, colon and breast cancer cell line. Annals of Medical Research, 28(9), 1767-1770.[CrossRef]
  • Referans20. Boussabbeh, M., Ben Salem, I., Rjiba-Touati, K., Bouyahya, C., Neffati, F., Najjar, M. F., Bacha, H., Abid-Essefi, S. (2016). The potential effect of patulin on mice bearing melanoma cells: an anti-tumour or carcinogenic effect? Tumour Biology, 37(5), 6285-6295. [CrossaRef]
  • Referans21. Wu, T. S., Liao, Y. C., Yu, F. Y., Chang, C. H., Liu, B. H. (2008). Mechanism of patulin-induced apoptosis in human leukemia cells (HL-60). Toxicology Letters, 183(1-3), 105-111.[CrossRef]
  • Referans22. Luft, P., Oostingh, G. J., Gruijthuijsen, Y., Horejs-Hoeck, J., Lehmann, I., Duschl, A. (2008). Patulin influences the expression of Th1/Th2 cytokines by activated peripheral blood mononuclear cells and T cells through depletion of intracellular glutathione. Environmental Toxicology, 23(1), 84-95.[CrossRef]
  • Referans23. Jin, H., Yin, S., Song, X., Zhang, E., Fan, L., Hu, H. (2016). p53 activation contributes to patulin-induced nephrotoxicity via modulation of reactive oxygen species generation. Scientific Reports, 6, 24455. [CrossRef]
  • Referans24. Zhou, S. M., Jiang, L. P., Geng, C. Y., Cao, J., & Zhong, L. F. (2010). Patulin-induced oxidative DNA damage and p53 modulation in HepG2 cells. Toxicon, 55(2-3), 390-395. [CrossRef]
  • Referans25. Saxena, N., Ansari, K. M., Kumar, R., Dhawan, A., Dwivedi, P. D., Das, M. (2009). Patulin causes DNA damage leading to cell cycle arrest and apoptosis through modulation of Bax, p(53) and p(21/WAF1) proteins in skin of mice. Toxicology and Applied Pharmacology, 234(2), 192-201. [CrossRef]

IN VITRO ANTITUMOR ACTIVITY OF PATULIN ON LİVER AND LUNG CANCER CELL LINES

Yıl 2022, Cilt: 46 Sayı: 2, 393 - 404, 29.05.2022
https://doi.org/10.33483/jfpau.1078080

Öz

Objective: Patulin is a mycotoxin produced by some molds, mainly Aspergillus and Penicillium, and is responsible for mycotoxicosis in animals and humans. Although there are still no detailed data on the antitumor activity of patulin, some reports indicate that it induces cellular apoptosis and toxicity. Our aim in this study is to determine the antiproliferative and antimigration effects of patulin on human liver and lung cancer cells and its effect on gene expressions in apoptotic pathways.
Material and Method: HEP3B and A549 cell lines were used to determine the efficacy of patulin in the treatment of liver and lung cancers. Cell lines were exposed to increasing concentrations of patulin (1, 2.5, 5, 10, 25, 50, and 100 µM), followed by determination of cell viability, cell migration, and apoptosis by MTS assay, wound healing assay, and RT-PCR assays.
Result and Discussion: Patulin caused a dose-dependent significant decrease in cell viability and cell migration in both cancer cell lines. It also promoted the induction of apoptosis in both cell lines. Our results showed that patulin can significantly reduce tumor growth in human liver and lung cancer cells in vitro.

Proje Numarası

TSG-2020-2087

Kaynakça

  • Referans1. Ismaiel, A.A., Papenbrock, J. (2017). Effect of patulin from penicillium vulpinum on the activity of glutathione-S-transferase and selected antioxidative enzymes in Maize. International Journal of Environmental Research and Public Health, 14(7), 825. [CrossRef]
  • Referans2. Puel, O., Galtier, P., Oswald, I.P. (2010). Biosynthesis and toxicological effects of patulin. Toxins (Basel), 2(4), 613-631. [CrossRef]
  • Referans3. Liu, B.H., Yu, F.Y., Wu, T.S., Li, S.Y., Su, M.C., Wang, M.C., Shih, S.M. (2003). Evaluation of genotoxic risk and oxidative DNA damage in mammalian cells exposed to mycotoxins, patulin and citrinin. Toxicology and Applied Pharmacology, 191(3), 255-63. [CrossRef]
  • Referans4. IARC. Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to Humans. IARC; Lyon, France: 1986. Some Naturally Occurring and Synthetic Food Components, Furocoumarins and Ultraviolet Radiation; pp. 83-98.
  • Referans5. Abastabar, M., Akbari, A., Akhtari, J., Hedayati, M. T., Shokohi, T., Mehrad-Majd, H., Ghalehnoei, H., Ghasemi, S. (2017). In vitro antitumor activity of patulin on cervical and colorectal cancer cell lines. Current Medical Mycology, 3(1), 25-29.[CrossRef]
  • Referans6. Boussabbeh, M., Ben Salem, I., Prola, A., Guilbert, A., Bacha, H., Abid-Essefi, S., Lemaire, C. (2015). Patulin induces apoptosis through ROS-mediated endoplasmic reticulum stress pathway. Toxicological Sciences, 144(2), 328-337.[CrossRef]
  • Referans7. Pal, S., Singh, N., & Ansari, K. M. (2017). Toxicological effects of patulin mycotoxin on the mammalian system: an overview. Toxicology Research, 6(6), 764–771. [CrossRef]
  • Referans8. Liu, J., Liu, Q., Han, J., Feng, J., Guo, T., Li, Z., Min, F., Jin, R., Peng, X. (2021). N-Acetylcysteine inhibits patulin-induced apoptosis by affecting ROS-mediated oxidative damage pathway. Toxins, 13(9), 595. [CrossRef]
  • Referans9. Barltrop, J.A., Owen, T.C., Cory, A.H., Cory, J.G. (1991). 5-(3- carboxymethoxyphenyl)-2-(4,5-dimethylthiazolyl)-3- (4-sulfophenyl) tetrazolium, inner salt (MTS) and related analogs of 3-(4,5-dimethylthiazolyl)-2,5- diphenyltetrazolium bromide (MTT) reducing to purple water-soluble formazans as cell-viability indicators. Bioorganic & Medicinal Chemistry Letters, 1(11), 611-614. [CrossRef]
  • Referans10. Liang, C.C., Park, A., Guan, J.L. (2007). In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro. Nature Protocols, 2(2), 329-333. [CrossRef]
  • Referans11. Demirci, S., Doğan, A., Türkmen, N. B., Telci, D., Rizvanov, A. A., Şahin, F. (2017). Schiff base-Poloxamer P85 combination demonstrates chemotherapeutic effect on prostate cancer cells in vitro. Biomedicine & Pharmacotherapy, 86, 492-501. [CrossRef]
  • Referans12. Andersen, B., Smedsgaard, J., Frisvad, J.C. (2004). Penicillium expansum: Consistent production of patulin, chaetoglobosins, and other secondary metabolites in culture and their natural occurrence in fruit products. Journal of Agricultural and Food Chemistry, 52(8), 2421-2428. [CrossRef]
  • Referans13. Kwon, O., Soung, N. K., Thimmegowda, N. R., Jeong, S. J., Jang, J. H., Moon, D. O., Chung, J. K., Lee, K. S., Kwon, Y. T., Erikson, R. L., Ahn, J. S., Kim, B. Y. (2012). Patulin induces colorectal cancer cells apoptosis through EGR-1 dependent ATF3 up-regulation. Cellular Signalling, 24(4), 943-950. [CrossRef]
  • Referans14. Liu, B. H., Wu, T. S., Yu, F. Y., Wang, C. H. (2006). Mycotoxin patulin activates the p38 kinase and JNK signaling pathways in human embryonic kidney cells. Toxicological Sciences, 89(2), 423-430. [CrossRef]
  • Referans15. Guo, X., Dong, Y., Yin, S., Zhao, C., Huo, Y., Fan, L. (2013). Patulin induces pro-survival functions via autophagy inhibition and p62 accumulation. Cell Death Disease, 4(10), e822. [CrossRef]
  • Referans16. Ferrer, E., Juan-García, A., Font, G., Ruiz, M. J. (2009). Reactive oxygen species induced by beauvericin, patulin and zearalenone in CHO-K1 cells. Toxicology In Vitro, 23(8), 1504-1509.[CrossRef]
  • Referans17. Alves, I., Oliveira, N., Laires, A., Rodrigues, A., Rueff, J. (2000). Induction of micronuclei and chromosomal aberrations by the mycotoxin patulin in mammalian cells: role of ascorbic acid as a modulator of patulin clastogenicity. Mutagenesis, 15(3), 229-34. [CrossRef]
  • Referans18. Burghardt, R. C., Barhoumi, R., Lewis, E. H., Bailey, R. H., Pyle, K. A., Clement, B. A., Phillips, T. D. (1992). Patulin-induced cellular toxicity: a vital fluorescence study. Toxicology and Applied Pharmacology, 112(2), 235-244. [CrossRef]
  • Referans19. Turkmen, N. B., Yuce, H., Ozek, D. A., Aslan, S., Yasar, S., Unuvar, S. (2021). Dose dependent cytotoxic activity of patulin on neuroblastoma, colon and breast cancer cell line. Annals of Medical Research, 28(9), 1767-1770.[CrossRef]
  • Referans20. Boussabbeh, M., Ben Salem, I., Rjiba-Touati, K., Bouyahya, C., Neffati, F., Najjar, M. F., Bacha, H., Abid-Essefi, S. (2016). The potential effect of patulin on mice bearing melanoma cells: an anti-tumour or carcinogenic effect? Tumour Biology, 37(5), 6285-6295. [CrossaRef]
  • Referans21. Wu, T. S., Liao, Y. C., Yu, F. Y., Chang, C. H., Liu, B. H. (2008). Mechanism of patulin-induced apoptosis in human leukemia cells (HL-60). Toxicology Letters, 183(1-3), 105-111.[CrossRef]
  • Referans22. Luft, P., Oostingh, G. J., Gruijthuijsen, Y., Horejs-Hoeck, J., Lehmann, I., Duschl, A. (2008). Patulin influences the expression of Th1/Th2 cytokines by activated peripheral blood mononuclear cells and T cells through depletion of intracellular glutathione. Environmental Toxicology, 23(1), 84-95.[CrossRef]
  • Referans23. Jin, H., Yin, S., Song, X., Zhang, E., Fan, L., Hu, H. (2016). p53 activation contributes to patulin-induced nephrotoxicity via modulation of reactive oxygen species generation. Scientific Reports, 6, 24455. [CrossRef]
  • Referans24. Zhou, S. M., Jiang, L. P., Geng, C. Y., Cao, J., & Zhong, L. F. (2010). Patulin-induced oxidative DNA damage and p53 modulation in HepG2 cells. Toxicon, 55(2-3), 390-395. [CrossRef]
  • Referans25. Saxena, N., Ansari, K. M., Kumar, R., Dhawan, A., Dwivedi, P. D., Das, M. (2009). Patulin causes DNA damage leading to cell cycle arrest and apoptosis through modulation of Bax, p(53) and p(21/WAF1) proteins in skin of mice. Toxicology and Applied Pharmacology, 234(2), 192-201. [CrossRef]
Toplam 25 adet kaynakça vardır.

Ayrıntılar

Birincil Dil Türkçe
Konular Eczacılık ve İlaç Bilimleri
Bölüm Araştırma Makalesi
Yazarlar

Hande Yüce 0000-0003-2907-2019

Neşe Başak Türkmen 0000-0001-5566-8321

Selinay Şenkal 0000-0002-6897-7307

Dilan Aşkın Özek 0000-0001-9075-4807

Ezgi Bulut 0000-0001-5842-6088

Ayşegül Doğan 0000-0003-4160-2270

Songül Ünüvar 0000-0001-8454-490X

Proje Numarası TSG-2020-2087
Yayımlanma Tarihi 29 Mayıs 2022
Gönderilme Tarihi 24 Şubat 2022
Kabul Tarihi 19 Nisan 2022
Yayımlandığı Sayı Yıl 2022 Cilt: 46 Sayı: 2

Kaynak Göster

APA Yüce, H., Başak Türkmen, N., Şenkal, S., Aşkın Özek, D., vd. (2022). PATULİNİN KARACİĞER VE AKCİĞER KANSERİ HÜCRE HATLARI ÜZERİNDEKİ İN VİTRO ANTİTÜMÖR AKTİVİTESİ. Journal of Faculty of Pharmacy of Ankara University, 46(2), 393-404. https://doi.org/10.33483/jfpau.1078080
AMA Yüce H, Başak Türkmen N, Şenkal S, Aşkın Özek D, Bulut E, Doğan A, Ünüvar S. PATULİNİN KARACİĞER VE AKCİĞER KANSERİ HÜCRE HATLARI ÜZERİNDEKİ İN VİTRO ANTİTÜMÖR AKTİVİTESİ. Ankara Ecz. Fak. Derg. Mayıs 2022;46(2):393-404. doi:10.33483/jfpau.1078080
Chicago Yüce, Hande, Neşe Başak Türkmen, Selinay Şenkal, Dilan Aşkın Özek, Ezgi Bulut, Ayşegül Doğan, ve Songül Ünüvar. “PATULİNİN KARACİĞER VE AKCİĞER KANSERİ HÜCRE HATLARI ÜZERİNDEKİ İN VİTRO ANTİTÜMÖR AKTİVİTESİ”. Journal of Faculty of Pharmacy of Ankara University 46, sy. 2 (Mayıs 2022): 393-404. https://doi.org/10.33483/jfpau.1078080.
EndNote Yüce H, Başak Türkmen N, Şenkal S, Aşkın Özek D, Bulut E, Doğan A, Ünüvar S (01 Mayıs 2022) PATULİNİN KARACİĞER VE AKCİĞER KANSERİ HÜCRE HATLARI ÜZERİNDEKİ İN VİTRO ANTİTÜMÖR AKTİVİTESİ. Journal of Faculty of Pharmacy of Ankara University 46 2 393–404.
IEEE H. Yüce, “PATULİNİN KARACİĞER VE AKCİĞER KANSERİ HÜCRE HATLARI ÜZERİNDEKİ İN VİTRO ANTİTÜMÖR AKTİVİTESİ”, Ankara Ecz. Fak. Derg., c. 46, sy. 2, ss. 393–404, 2022, doi: 10.33483/jfpau.1078080.
ISNAD Yüce, Hande vd. “PATULİNİN KARACİĞER VE AKCİĞER KANSERİ HÜCRE HATLARI ÜZERİNDEKİ İN VİTRO ANTİTÜMÖR AKTİVİTESİ”. Journal of Faculty of Pharmacy of Ankara University 46/2 (Mayıs 2022), 393-404. https://doi.org/10.33483/jfpau.1078080.
JAMA Yüce H, Başak Türkmen N, Şenkal S, Aşkın Özek D, Bulut E, Doğan A, Ünüvar S. PATULİNİN KARACİĞER VE AKCİĞER KANSERİ HÜCRE HATLARI ÜZERİNDEKİ İN VİTRO ANTİTÜMÖR AKTİVİTESİ. Ankara Ecz. Fak. Derg. 2022;46:393–404.
MLA Yüce, Hande vd. “PATULİNİN KARACİĞER VE AKCİĞER KANSERİ HÜCRE HATLARI ÜZERİNDEKİ İN VİTRO ANTİTÜMÖR AKTİVİTESİ”. Journal of Faculty of Pharmacy of Ankara University, c. 46, sy. 2, 2022, ss. 393-04, doi:10.33483/jfpau.1078080.
Vancouver Yüce H, Başak Türkmen N, Şenkal S, Aşkın Özek D, Bulut E, Doğan A, Ünüvar S. PATULİNİN KARACİĞER VE AKCİĞER KANSERİ HÜCRE HATLARI ÜZERİNDEKİ İN VİTRO ANTİTÜMÖR AKTİVİTESİ. Ankara Ecz. Fak. Derg. 2022;46(2):393-404.

Kapsam ve Amaç

Ankara Üniversitesi Eczacılık Fakültesi Dergisi, açık erişim, hakemli bir dergi olup Türkçe veya İngilizce olarak farmasötik bilimler alanındaki önemli gelişmeleri içeren orijinal araştırmalar, derlemeler ve kısa bildiriler için uluslararası bir yayım ortamıdır. Bilimsel toplantılarda sunulan bildiriler supleman özel sayısı olarak dergide yayımlanabilir. Ayrıca, tüm farmasötik alandaki gelecek ve önceki ulusal ve uluslararası bilimsel toplantılar ile sosyal aktiviteleri içerir.