HCT116 İnsan Kolon Kanseri Hücrelerinde Endemik Phlomis Ekstraktlarının Antikanser Aktivitesi

Amaç: Önceki çalışmalar, Phlomis russeliana'nın çeşitli kanser hücre hatlarına karşı sitotoksik etkiler gösterdiğini bildirmiştir; ancak, HCT-116 kanser hücreleri üzerindeki anti kanser aktivitesi henüz araştırılmamıştır. Bu nedenle, bu çalışma Phlomis russeliana'nın HCT-116 insan kolon kanseri hücre dizisi ve HUVEC normal hücre dizisine karşı anti-kanser özelliklerini araştırmak üzere tasarlanmıştır. Gereç ve Yöntemler: HCT-116 ve HUVEC hücreleri Phlomis russeliana’nın farklı konsantrasyonları (2, 4, 6, 8 ve 10 mg/ml) ile muamele edildi ve hücre canlılığı MTT yöntemi ile değerlendirildi. Phlomis russeliana'nın anti-migrasyon ve anti-kolonijenik etkileri, sırasıyla yara iyileşmesi ve koloni oluşumu analizleri ile değerlendirildi. Kolorimetrik Elisa kitleri ile toplam antioksidan durumunun (TAS), toplam oksidan durumunun (TOS) ve kaspaz-3 aktivasyonunun kantitatif tespiti yapıldı. Bulgular: Phlomis russeliana, konsantrasyona bağlı olarak HCT-116 hücrelerinin canlılığını önemli ölçüde azalttı ve normal HUVEC'lere karşı daha zayıf toksisite gösterdi. Phlomis russeliana, HCT-116 hücrelerinin göçünü ve koloni oluşturma potansiyelini önemli ölçüde engelledi. Phlomis russeliana ile tedaviden sonra kaspaz-3 aktivasyonunda önemli bir artış gözlendi. Phlomis russeliana, HCT-116 hücrelerinde TAS ve TOS seviyesini önemli ölçüde etkilemedi. Sonuç: Bu sonuçlar; Phlomis russeliana'nın koloni oluşumunun baskılanması, migrasyonun engellenmesi ve kaspaz-3 aktivasyonunun indüklenmesi yoluyla insan kolon kanseri hücrelerinde anti-kanser aktivitesi gösterdiğini ortaya koydu. Phlomis russeliana, kansere karşı yeni anti-kanser ajanlarının geliştirilmesi için umut verici bir kaynak olabilir.

Anahtar Kelimeler:

Phlomis russeliana, HCT-116 hücreleri, migrasyon, kaspaz-3 aktivasyonu, koloni oluşumu

Anticancer Activity of Endemic Phlomis Extracts in HCT116 Human Colon Cancer Cells

Aim: Previous studies have reported that Phlomis russeliana shows cytotoxic effects against several cancer cell lines; however, its anti cancer activity on HCT-116 cancer cells has not yet been investigated. Therefore, the present study is designed to explore anti-cancer properties of Phlomis russeliana against HCT-116 human colon cancer cell line and HUVEC normal cell line. Material and Methods: HCT-116 cells and HUVECs treated with different concentrations of Phlomis russeliana (2, 4, 6, 8 and 10 mg/ml) and cell viability evaluated by the MTT assay. Anti-migratory and anti-colonigenic effects of Phlomis russeliana were assessed with wound healing and colony formation assays respectively. Quantitative determination of total antioxidant status (TAS), total oxidant status (TOS) and caspase-3 activation were performed with colorimetric Elisa kits. Results: Phlomis russeliana significantly decreased cell viability of HCT-116 cells in a concentration dependent and showed weaker toxicity against normal HUVECs. Pholomis russeliana significantly inhibited migration and colony formation potential of HCT-116 cells. A significant increase in caspase-3 activation was observed after treatment with Phlomis russeliana. Phlomis russeliana did not significantly affect the TAS and TOS level in HCT-116 cells. Conclusion: These results revealed that Phlomis russeliana showed anti-cancer activity in human colon cancer cells, through the suppression of colony formation, inhibition of migration and induction of caspase-3 activation. Phlomis russeliana, could be a promising source for the development of new anti-cancer agents against cancer.

Keywords:

Phlomis russeliana, HCT-116 cells, migration, caspase-3 activation, colony formation,

PDF

___

Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, Bray F, et al. Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. CA Cancer J Clin. 2021; 71(3): 209-49.
Saif MW, Chu E. Biology of colorectal cancer. Cancer J. 2010; 16(3): 196-1.
Bagheri E, Hajiaghaalipour F, Nyamathulla S, Salehen NA. Ethanolic extract of Brucea javanica inhibit proliferation of HCT-116 colon cancer cells via caspase activation. RSC Adv. 2018; 8(2): 681-89.
Chen MB, Zhang Y, Wei MX, Shen W, Wu XY, Yao C, et al. Activation of AMP-activated protein kinase (AMPK) mediates plumbagin-induced apoptosis and growth inhibition in cultured human colon cancer cells. Cell Signal. 2013; 25(10): 1993-2.
Kirmizibekmez H, Montoro P, Piacente S, Pizza C, Dönmez A, Caliş I. Identification by HPLC-PAD-MS and quantification by HPLC-PAD of phenylethanoid glycosides of five Phlomis species. Phytochem Anal. 2005; 16(1): 1-6.
Amor LBI, Boubaker J, Sgaier BM, Skandrani I, Bhouri W, Neffati A, et al. Phytochemistry and biological activities of Phlomis species. J Ethnopharmacol. 2009; 125(2): 183-2.
Demirci B, Toyota M, Demirci F, Dadandi MY, Can Baser KH. Anticandidal pimaradiene diterpene from Phlomis essential oils. Comptes Rendus Chim. 2009; (12): 612-21.
Saracoglu I, Inoue M, Calis I, Ogihara Y. Studies on constituents with cytotoxic and cytostatic activity of two Turkish medicinal plants Phlomis armeniaca and Scutellaria salviifolia. Biol Pharm Bull. 1995; 18(10): 1396-1400.
Heilmann J, Calis I, Kirmizibekmez H, Schühly W, Harput S, Sticher O. Radical scavenger activity of phenylethanoid glycosides in FMLP stimulated human polymorphonuclear leukocytes: structure-activity relationships. Planta Med. 2000; 66(8): 746-8.
Alpay M, Dulger G, Sahin, IE, Dulger B. Evaluating antimicrobial and antioxidant capacity of endemic Phlomis russeliana from Turkey and its antiproliferative effect on Human Caco-2 Cell Lines. Anais da Academia Brasileira de Ciências. 2019: 91(3): e20180404.
Ghaffari K, Ahmadi R, Saberi B, Moulavi P. Anti-proliferative effects of Ziziphus spina-christi and Phlomis russeliana leaf extracts on HEK293 and MCF-7 Cell Lines and Evaluation of Bax and Bcl-2 Genes Expression Level in MCF-7 Cells. Asian Pac J Cancer Prev. 2021; 22(S1): 81-7.
Sarkhail P, Sahranavard S, Nikan M, Gafari S, Eslami-Tehrani B. Evaluation of the cytotoxic activity of extracts from six species of Phlomis genus. J Appl Pharm Sci. 2017; 7(2): 180-4.
Sahranavard S, Naghibi F, Mosaddegh M, Esmaeili S, Sarkhail P, Taghvaei M, et al. Cytotoxic activities of selected medicinal plants from Iran and phytochemical evaluation of the most potent extract. Res Pharm Sci. 2009; 4(2): 133-7.
Ranjbar SH, Larijani B, Abdollahi M. A systematic review of Iranian medicinal plants useful in diabetes mellitus. Archives of Medical Science. 2008; 4(3): 285-92.
Digrak M, Alma MH, Ilcim A, Sen S. Antibacterial and antifungal effects of various commercial plant extracts. Pharmaceutical Biology. 1999; 37(3): 216-20.
Banerjee S, Uppal T, Strahan R, Dabral P, Verma SC. The Modulation of Apoptotic Pathways by Gammaherpesviruses. Front Microbiol. 2016; 7: 585.
Xiang L, He B, Liu Q, Hu D, Liao W, Li R, et al. Antitumor effects of curcumin on the proliferation, migration and apoptosis of human colorectal carcinoma HCT‑116 cells. Oncol Rep. 2020; 44(5): 1997-2008.
Zhou M, Liu X, Li Z, Huang Q, Li F, Li CY. Caspase-3 regulates the migration, invasion and metastasis of colon cancer cells. Int J Cancer. 2018; 143(4): 921-30.
Catalano M, D'Alessandro G, Lepore F, Corazzari M, Caldarola S, Valacca C, et al. Autophagy induction impairs migration and invasion by reversing EMT in glioblastoma cells. Mol Oncol. 2015; 9(8): 1612-25.
Liang ZE, Yi YJ, Guo YT, Wang RC, Hu QL, Xiong XY. Inhibition of migration and induction of apoptosis in LoVo human colon cancer cells by polysaccharides from Ganoderma lucidum. Mol Med Rep. 2015; 12(5): 7629-36.
Menyhart O, Harami-Papp H, Sukumar S, Schafer R, Magnani L, de Barrios O, et al. Guidelines for the selection of functional assays to evaluate the hallmarks of cancer. Biochim Biophys Acta. 2016; 1866(2): 300-19.
Raeisi F, Raeisi E, Heidarian E, Shahbazi-Gahroui D, Lemoigne Y. Bromelain ınhibitory effect on colony formation: an ın vitro study on human AGS, PC3, and MCF7 cancer cells. J Med Signals Sens. 2019; 9(4): 267-3.
Kim A, Ma JY. Rhaponticin decreases the metastatic and angiogenic abilities of cancer cells via suppression of the HIF‑1α pathway. Int J Oncol. 2018; 53(3): 1160-70.
Du XF, Zhang LL, Zhang DZ, Yang L, Fan YY, Dong SP. Clinical significance of serum total oxidant/antioxidant status in patients with operable and advanced gastric cancer. Onco Targets Ther. 2018; 11: 6767-75.