ECHINOCOCCUS GRANULOSUS SENSU STRICTO KAYNAKLI HİDATİK KİST SIVISI UYGULAMASININ CACO-2 HÜCRE HATTINDA EPİTELYAL-MEZENKİMAL GEÇİŞ VE APOPTOZ ÜZERİNE ETKİSİ

Kistik ekinokokkoz (KE), Echinococcus granulosus sensu lato'nun larva formunun neden olduğu zoonotik bir enfeksiyondur. Yapılan çalışmalar E. granulosus enfeksiyonu ile kanser arasında doğrudan ve/veya dolaylı bir ilişki olduğu öne sürmüş; ancak, elde edilen sonuçlar farklı hücre kültürü ve/veya hayvan modellerinde hem anti-kanserojen hem de kanserojen etkisi olabileceğini göstermiştir. İnsan kolorektal adenokarsinom (Caco-2) hücrelerine etkisi daha önce değerlendirilmemiştir. Bu çalışmanın amacı, hidatik kist sıvısı uygulamasının bazı apoptotik genlerin (BCL-2, p53 ve BAX) ve epitelyal-mezenkimal geçiş (EMT) genlerinin (e-kaderin ve vimentin) ekspresyonu ve hücre proliferasyonu üzerine etkisini değerlendirerek Caco-2 hücre hattında olası anti-kanserojen veya kanserojen etkisini moleküler düzeyde aydınlatmaktır. Hidatik kist sıvısı uygulamasının sonrasında hücre proliferasyonu, apoptotik genler ve EMT gen ekspresyonu üzerindeki etkisini değerlendirmek için hücre proliferasyon analizi (XTT ile) ve gerçek zamanlı polimeraz zincir reaksiyonu (RT-PCR) yapılmıştır. Uygulama sonrasında uygulama dozu ile orantılı olarak hücre proliferasyonunda istatistiksel olarak anlamlı bir artış tespit edilmiştir. BAX ve p53 gen ifadelerinde doza bağlı azalma ve BCL-2 gen ifadesinde artış tespit edilmiştir. Bunun yanı sıra hücre kültüründe EMT gözlenmiş ve e-kaderin (CDH1) ve vimentin ekspresyonları ile moleküler düzeyde doğrulanmıştır. Bu çalışma ile hidatik kist sıvısının, Caco-2 hücre hattına uygulanması hücre proliferasyonunu doğrudan arttırdığı ve Caco-2 hücre hattının apoptoza karşı çok daha dirençli ve metastatik hale gelmesine neden olduğu tespit edilmiştir. Bu çalışma, ilk kez Caco-2 hücre hattından apoptotik yolakta hidatik kist sıvının olası mekanizmasına ışık tutmaktadır.

Anahtar Kelimeler:

apoptoz, Echinococcus granulosus, epitelyal-mezenkimal geçiş, kanser, kist hidatik sıvısı

The Effect of Echinococcus granulosus sensu stricto-Derived Hydatid Cyst Fluid Application on Epithelial-Mesenchymal Transition and Apoptosis on Caco-2 Cell Line

Cystic echinococcosis (CE) is a zoonotic infection caused by the larval form of Echinococcus granulosus sensu lato. Studies suggested a direct and/or indirect relationship between E. granulosus infection and cancer; however, results showed both anti-carcinogenic and carcinogenic effects in different cell culture and/or animal models. Its effect on human colorectal adenocarcinoma (Caco-2) cells has not been previously evaluated. The aim of this study was elucidate the molecular mechanism underlying the possible anti-carcinogenic or carcinogenic effect of hydatid cyst fluid on Caco-2 cell line at the molecular level via evaluating the effect of its application on cell proliferation and expression of some apoptotic genes (BCL-2, p53 and BAX) and epithelial-mesenchymal transition (EMT) genes (e-cadherin and vimentin). Cell proliferation analysis (using XTT) and real-time polymerase chain reaction (RT-PCR) were performed to evaluate the effect of hydatid cyst fluid on cell proliferation and apoptotic and EMT gene expression, respectively. After the application, a statistically significant increase in cell proliferation was detected correlated with the application dose. A dose-dependent decrease in BAX and p53 gene expressions and an increase in BCL-2 gene expression were determined. EMT is also observed and confirmed via e-cadherin (CDH1) and vimentin expression levels. With this study, the application of hydatid cyst fluid to Caco-2 cells directly increases cell proliferation. Thus, it caused the Caco-2 cell line to become much more resistant to apoptosis and prone to metastasis. This study sheds light on the possible mechanism of hydatid cyst fluid in the apoptotic pathway from the Caco-2 cell line for the first time.

Keywords:

apoptosis, cancer, Echinococcus granulosus, epithelial-mesenchymal transition, hydatid cyst fluid,

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Agudelo Higuita NI, Brunetti E, McCloskey C. Cystic echinococcosis. J Clin Microbiol. 2016;54(3):518-23.
Akgül H, Tez M, Unal AE, Keşkek M, Sayek I, Ozçelik T. Echinococcus against cancer: why not? Cancer. 2003;98(9):1999-2000.
Alvarez Errico D, Medeiros A, Míguez M, et al. O-glycosylation in Echinococcus granulosus: identification and characterization of the carcinoma-associated Tn antigen. Exp Parasitol. 2001;98(2):100-9.
Atayde VD, Jasiulionis MG, Cortez M, Yoshida N. A recombinant protein based on Trypanosoma cruzi surface molecule gp82 induces apoptotic cell death in melanoma cells. Melanoma Res. 2008;18(3):172-83.
Berriel E, Russo S, Monin L, et al. Antitumor activity of human hydatid cyst fluid in a murine model of colon cancer. ScientificWorldJournal. 2013;2013:230176.
Daneshpour S, Kefayat AH, Mofid MR, Rostami Rad S, Yousofi Darani H. Effect of hydatid cyst fluid antigens on induction of apoptosis on breast cancer cells. Adv Biomed Res. 2019;8:27.
Daneshpour S, Rostamirad S, Kefayat A, Mofid M, Safavi A, Darani HY. Identifying the most effective hydatid cyst fluid fraction for anticancer vaccination of 4T1 breast tumor-bearing mice. Int J Prev Med. 2019;10:143.
Deplazes P, Rinaldi L, Alvarez Rojas CA, et al. Global distribution of alveolar and cystic echinococcosis. Adv Parasitol. 2017;95:315-493.
Eckert J, Gemmell MA, Meslin F-X, Pawlowski ZS(ed.), WHO/OIE manual on echinococcosis in humans and animals: a public health problem of global concern. World Organisation for Animal Health, Paris, France (2001).
Guan W, Zhang X, Wang X, Lu S, Yin J, Zhang J. Employing parasite against cancer: a lesson from the canine tapeworm Echinococcus granulocus. Front Pharmacol. 2019;10:1137.
Kanapathipillai M. Treating p53 mutant aggregation-associated cancer. Cancers (Basel). 2018;10(6).
Kang Y, Massagué J. Epithelial-mesenchymal transitions: twist in development and metastasis. Cell. 2004;118(3):277-9.
Khodapasand E, Jafarzadeh N, Farrokhi F, Kamalidehghan B, Houshmand M. Is Bax/Bcl-2 ratio considered as a prognostic marker with age and tumor location in colorectal cancer? Iran Biomed J. 2015;19(2):69-75.
Kosmider B, Wojcik I, Osiecka R, et al. Enhanced P53 and BAX gene expression and apoptosis in A549 cells by cis-Pt(II) complex of 3-aminoflavone in comparison with cis-DDP. Invest New Drugs. 2005;23(4):287-97.
Li H, Song T, Shao Y, Wen H. Cystic echinococcosis accompanied by hepatocellular carcinoma in a female herdsman. Int J Clin Exp Med. 2015;8(2):2985-8. Nakao M, Sako Y, Yokoyama N, Fukunaga M, Ito A. Mitochondrial genetic code in cestodes. Mol Biochem Parasitol. 2000;111(2):415-24.
Noguti J, Barbisan LF, Cesar A, Dias Seabra C, Choueri RB, Ribeiro DA. Review: In vivo models for measuring placental glutatione-S-transferase (GST-P 7-7) levels: a suitable biomarker for understanding cancer pathogenesis. In Vivo. 2012;26(4):647-50.
Oikonomopoulou K, Brinc D, Kyriacou K, Diamandis EP. Infection and cancer: revaluation of the hygiene hypothesis. Clin Cancer Res. 2013;19(11):2834-41.
Oikonomopoulou K, Yu H, Wang Z, et al. Association between Echinococcus granulosus infection and cancer risk - a pilot study in Cyprus. Clin Chem Lab Med. 2016;54(12):1955-61.
Pawlowski Z, Eckert J, Vuitton DA, et al., editors. Echinococcosis in humans: clinical aspects, diagnosis and treatment2001.
Perlman H, Zhang X, Chen MW, Walsh K, Buttyan R. An elevated bax/bcl-2 ratio corresponds with the onset of prostate epithelial cell apoptosis. Cell Death Differ. 1999;6(1):48-54.
Pisani P, Parkin DM, Muñoz N, Ferlay J. Cancer and infection: estimates of the attributable fraction in 1990. Cancer Epidemiol Biomarkers Prev. 1997;6(6):387-400.
Raisova M, Hossini AM, Eberle J, et al. The Bax/Bcl-2 ratio determines the susceptibility of human melanoma cells to CD95/Fas-mediated apoptosis. J Invest Dermatol. 2001;117(2):333-40.
Ranasinghe SL, McManus DP. Echinococcus granulosus: Cure for cancer revisited. Front Med (Lausanne). 2018;5:60.
Rook GA, Dalgleish A. Infection, immunoregulation, and cancer. Immunol Rev. 2011;240(1):141-59.
Sayek I. Kist Hidatik Hastalığı: Klinik Yönleri. Altıntaş N TR, Çoker A., editor. İzmir: Hidatidoloji Derneği Yayınları; (2004).
Tamarozzi F, Akhan O, Cretu CM, et al. Prevalence of abdominal cystic echinococcosis in rural Bulgaria, Romania, and Turkey: a cross-sectional, ultrasound-based, population study from the HERACLES project. Lancet Infect Dis. 2018;18(7):769-78.
Topluoglu S. Current situation report of cystic echinococcosis in Turkey. Turk Hij Den Biyol Derg. 2020;77(70):1-52.
Yasen A, Wang M, Ran B, et al. Echinococcus granulosus protoscoleces promotes proliferation and invasion of hepatocellular carcinoma cells. Cytotechnology. 2021;73(1):13-22.
Yousofi Darani H, Soozangar N, Khorami S, Taji F, Yousofi M, Shirzad H. Hydatid cyst protoscolices induce cell death in WEHI-164 fibrosarcoma cells and inhibit the proliferation of baby hamster kidney fibroblasts in vitro. J Parasitol Res. 2012;2012:304183.
Zhao XJ, Li H, Chen H, et al. Expression of e-cadherin and beta-catenin in human esophageal squamous cell carcinoma: relationships with prognosis. World J Gastroenterol. 2003;9(2):225-32.