Investigation of the Synergistic Effect of Allium Polyanthum Schult Extract and Docetaxel on Apoptosis-Related Genes in Colorectal Cancer Cells and Bioinformatics Analysis

Zuhal Tunçbilek; Gonca Kabak; Yavuz Siliğ

doi:10.7197/cmj.1601162

Araştırma Makalesi

Allium Polyanthum Schult Ekstraktı ve Dosetakselin Kolorektal Kanser Hücrelerindeki Apoptozla İlgili Genler Üzerindeki Sinerjik Etkisinin Araştırılması

Yıl 2024, Cilt: 46 Sayı: 4, 280 - 291, 31.12.2024

Zuhal Tunçbilek , Gonca Kabak , Yavuz Siliğ

https://doi.org/10.7197/cmj.1601162

Öz

Amaç: Kolorektal kanser (CRC), yüksek mortalite ve yaygın metastazlı bir kanser türüdür. CRC'nin erken teşhis edildiği durumlarda kemoterapi ve cerrahi uygulamalar esastır. Docetaxel, taksan grubunda bir antikanser ajandır. Ayrıca, tıbbi aktiviteye sahip bitkilerin CRC üzerinde koruyucu bir etkiye sahip olduğu bilinmektedir. Amaç, Allium polyanthum Schult bitki özütü ve bu bitki özütünün dosetaksel ile kombinasyonu ile tedavi edilen kolorektal kanser hücrelerinde apoptozisle ilişkili genlerin ifade düzeylerini belirlemekti.
Gereç ve Yöntem: Sadece Allium polyanthum Schult bitki ekstresi ve bu bitki ekstresiyle birlikte docetaxel uygulanan CRC hücre hattı HT-29 ve sağlıklı kolon hücre hattı CCD-18Co'da apoptozda rol oynayan kaspaz-2 (CASP2), nükleer faktör NF-kappa-B1 (NFKB1), apoptoz düzenleyici BAX ve proto-onkogen MYC genlerinin ekspresyon analizleri, gerçek zamanlı polimeraz zincir reaksiyonu (RT-PCR) yöntemi kullanılarak gerçekleştirildi. Çeşitli veritabanları kullanılarak ilgili genlerin biyoinfarmatik analizi gerçekleştirildi.
Sonuçlar: CASP2, MYC, NFKB1 ve BAX gen ekspresyonu, Allium polyanthum Schult özütü ve docetaxel kombinasyonu ile tedavi edilen CRC hücrelerinde sağlıklı hücrelere kıyasla önemli ölçüde azaldı. Biyoenformatik analiz sonucunda, CASP2, MYC, NFKB1 ve BAX proteinlerinin birbirleriyle etkileşime girdiği ve genlerinin ekspresyon seviyelerinin hayatta kalma ile ilişkili olduğu bulundu. Ayrıca, CASP2 ve NFKB1'in metilasyon durumu, CRC'de epigenetik mekanizmaları etkileyerek protein seviyelerini değiştirme potansiyeline sahiptir.
Tartışma: Literatürdeki bilgilere göre, Allium türlerinin apoptotik yollardaki genleri etkilediği bildirilmiştir. Buna göre, Allium polyanthum Schult'un tek başına ve docetaksel ile kombinasyonu daha fazla çalışma ile desteklenmelidir.

Anahtar Kelimeler

Allium polyanthum Schult, apoptoz, docetaksel, gen ekspresyonu, kolorektal kanser

Kaynakça

1. GLOBOCAN. Cancer Today. 2022 Available from: https://gco.iarc.fr/. Access date: 06.12.2024.
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3. Hossain MS, Karuniawati H, Jairoun AA, et al. (2022). Colorectal cancer: a review of carcinogenesis, global epidemiology, current challenges, risk factors, preventive and treatment strategies. Cancers, 2022;14(7):1732. https://doi.org/10.3390/cancers14071732
4. Testa U, Pelosi E, Castelli G. Colorectal cancer: genetic abnormalities, tumor progression, tumor heterogeneity, clonal evolution and tumor-initiating cells. Medical Sciences, 2018;6(2):31. https://doi.org/10.3390/medsci6020031
5. Baldwin LM., Dobie, SA, Billingsley K, et al. Explaining black–white differences in receipt of recommended colon cancer treatment. J Natl Cancer Inst, 2005;97(16):1211-1220. https://doi.org/10.1093/jnci/dji241
6. Abraha AM, Ketema EB. Apoptotic pathways as a therapeutic target for colorectal cancer treatment. World J Gastrointest Oncol, 2016;8(8):583. https://doi.org/10.4251/wjgo.v8.i8.583
7. Kiraz Y, Adan A, Kartal Yandim M, Baran Y. Major apoptotic mechanisms and genes involved in apoptosis. Tumour Biol, (2016);37:8471-8486. https://doi.org/10.1007/s13277-016-5035-9
8. Hossain MS, Urbi Z, Sule A, Rahman KH. Andrographis paniculata (Burm. f.) Wall. ex Nees: a review of ethnobotany, phytochemistry, and pharmacology. Sci. World, 2014: (1):274905. https://doi.org/10.1155/2014/274905
9. Urbi Z, Zainuddin Z. Standardization of surface sterilization protocol of field grown Stevia rebaudiana prior to in vitro clonal propagation. Jurnal Teknologi, 2015;77(24). https://doi.org/10.11113/jt.v77.6722
10. Hossain MS, Urbi Z. Effect of naphthalene acetic acid on the adventitious rooting in shoot cuttings of Andrographis paniculata (Burm. f.) Wall. ex Nees: an important therapeutical herb. Int. J. Agron, 2016;(1):1617543. https://doi.org/10.1155/2016/1617543
11. Vuković S, Popović-Djordjević JB, Kostić AŽ, et al. Allium species in the Balkan region—Major metabolites, antioxidant and antimicrobial properties. Horticulturae, 2023;9(3):408. https://doi.org/10.3390/horticulturae9030408
12. Iwar K, Ochar K, Seo YA, Ha BK., Kim SH. Alliums as Potential Antioxidants and Anticancer Agents. Int. J. Mol. Sci., 2024;25(15):8079. https://doi.org/10.3390/ijms25158079
13. Mohammadi-Motlagh HR, Mostafaie A, Mansouri K. Anticancer and anti-inflammatory activities of shallot (Allium ascalonicum) extract. Archives of Medical Science, 2011;7(1):38-44. https://doi.org/10.5114/aoms.2011.20602
14. Poonthananiwatkul B, Lim RH, Howard RL, Pibanpaknitee P, Williamson EM. Traditional medicine use by cancer patients in Thailand. J. Ethnopharmacol., 2015;168:100-107. https://doi.org/10.1016/j.jep.2015.03.057
15. Alkan M, Güzel M, Akşit H, Bağdat RB, Alkan FR, Evlice E. Chemical components and insecticidal effects of essential oils from three lavender cultivars against adult Sitophilus granarius (L., 1758)(Coleoptera: Curculionidae). Turk. j. entomol, 2021;45(4):405-416. https://doi.org/10.16970/entoted.988985
16. Wang YJ, Huang Y, Anreddy N. Tea nanoparticle, a safe and biocompatible nanocarrier, greatly potentiates the anticancer activity of doxorubicin. Oncotarget, 2015;7(5):5877. https://doi.org/10.18632/oncotarget.6711
17. Chandrashekar DS, Bashel B, Balasubramanya SAH, et al. UALCAN: a portal for facilitating tumor subgroup gene expression and survival analyses. Neoplasia, 2017;19(8):649-658. https://doi.org/10.1016/j.neo.2017.05.002
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24. Wang Q, Shen X, Chen G, Du J. Drug resistance in colorectal cancer: from mechanism to clinic. Cancers, 2022;14(12):2928. https://doi.org/10.3390/cancers14122928
25. Kenmotsu H, Tanigawara Y. Pharmacokinetics, dynamics and toxicity of docetaxel: Why the Japanese dose differs from the Western dose. Cancer science, 2015;106(5):497-504. https://doi.org/10.1111/cas.12647
26. Beretta HV, Bannoud F, Insani M, et al. Relationships between bioactive compound content and the antiplatelet and antioxidant activities of six Allium vegetable species. Food Sci Biotechnol, 2017;55(2):266. https://doi.org/10.17113/ftb.55.02.17.4722
27. Suleria HAR, Butt MS, Anjum FM, Saeed F, Khalid N. Onion: Nature protection against physiological threats. Crit. Rev. Food Sci. Nutr., 2015;55(1):50-66. https://doi.org/10.1080/10408398.2011.646364
28. Sharifi-Rad J, Mnayer D, Tabanelli G, et al. Plants of the genus Allium as antibacterial agents: From tradition to pharmacy. Cell. Mol. Biol., 2016;62(9):57-68. https://doi.org/10.14715/cmb/2016.62.9.10
29. Abdelrahman M, Mahmoud HY, El-Sayed M, Tanaka S, Tran LS. Isolation and characterization of Cepa2, a natural alliospiroside A, from shallot (Allium cepa L. Aggregatum group) with anticancer activity. Plant Physiol Biochem, 2017;116:167-173. https://doi.org/10.1016/j.plaphy.2017.05.006
30. Kazemi M, Zolfaghari B, Keyvanlo Shahrestanaki M, Sadeghi Dinani M. Cytotoxic Effects of Allium affine Ledeb Butanolic Fraction on Breast and Ovary Cancer Cell Lines. J. Med. Plants 2017;16(64):83-90.
31. Abdel-Hady H, El-Sayed MM, Abdel-Gawad MM, El-Wakil EA, Abdel-Hameed ESS, Abdel-Lateef EES. LC-ESI-MS analysis, antitumor and antioxidant activities of methanolic extract of Egyptian Allium kurrat. J. Appl. Pharm. Sci., 2018;8(7):085-092. https://doi.org/10.7324/JAPS.2018.8714
32. Khazaei S, Esa NM, Ramachandran V, Hamid RA, Pandurangan AK, Etemad A, Ismail P. In vitro antiproliferative and apoptosis inducing effect of Allium atroviolaceum bulb extract on breast, cervical, and liver cancer cells. Front. pharmacol., 2017;8: 5. https://doi.org/10.3389/fphar.2017.00005
33. Khazaei S, Abdul Hamid R, Mohd Esa N, Ramachandran V, Aalam GTF, Etemad A, Ismail P. Promotion of HepG2 cell apoptosis by flower of Allium atroviolaceum and the mechanism of action. BMC Complement Altern Med, 2017;17:1-13. https://doi.org/10.1186/s12906-017-1594-6
34. Ivanova A, Mikhova B, Najdenski H, Tsvetkova I, Kostova I. Chemical composition and antimicrobial activity of wild garlic Allium ursinum of Bulgarian origin. Nat. Prod. Commun., 2009;4(8):1934578X0900400808. https://doi.org/10.1177/1934578X0900400808
35. Xu XY, Song GQ, Yu YQ, Ma HY, Ma L, Jin YN. Apoptosis and G2/M arrest induced by Allium ursinum (ramson) watery extract in an AGS gastric cancer cell line. Onco Targets Ther, 2013;779-783. https://doi.org/10.2147/OTT.S45865
36. Pârvu AE, Cătoi F, Deelawar S, Sarup D, Pârvu M. Anti-inflammatory effect of Allium ursinum. Not Sci Biol, 2014;6(1):20-26. https://doi.org/10.15835/nsb619252
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Investigation of the Synergistic Effect of Allium Polyanthum Schult Extract and Docetaxel on Apoptosis-Related Genes in Colorectal Cancer Cells and Bioinformatics Analysis

Yıl 2024, Cilt: 46 Sayı: 4, 280 - 291, 31.12.2024

Zuhal Tunçbilek , Gonca Kabak , Yavuz Siliğ

https://doi.org/10.7197/cmj.1601162

Öz

Objective: Colorectal cancer (CRC) is a type of cancer with high mortality and widespread metastasis. The aim was to determine the expression levels of apoptosis-related genes in CRC cells treated with Allium polyanthum Schult plant extract and the combination of this plant extract with docetaxel.
Material and Method: Expression analyses of caspase-2 (CASP2), nuclear factor NF-kappa-B1 (NFKB1), apoptosis regulator BAX and proto-oncogene MYC genes that play a role in apoptosis in the CRC cell line HT-29 and the healthy colon cell line CCD-18Co, which were treated with only Allium polyanthum Schult plant extract and docetaxel together with this plant extract, were performed using the real-time polymerase chain reaction (RT-PCR) method. Bioinformatics analysis of relevant genes was performed using various databases.
Results: CASP2, MYC, NFKB1 and BAX gene expression was significantly decreased in CRC cells treated with the combination of Allium polyanthum Schult extract and docetaxel compared to healthy cells. As a result of bioinformatics analysis, it was found that CASP2, MYC, NFKB1 and BAX proteins interact with each other and the expression levels of their genes are associated with survival. In addition, the methylation status of CASP2 and NFKB1 has the potential to change protein levels by affecting epigenetic mechanisms in CRC.
Discussion: According to the information in the literature, it has been reported that Allium species affect genes in apoptotic pathways. Accordingly, Allium polyanthum Schult alone and in combination with docetaxel should be supported by further studies.

Anahtar Kelimeler

Allium polyanthum Schult, apoptosis, colorectal cancer, docetaxel, gene expression

Kaynakça

1. GLOBOCAN. Cancer Today. 2022 Available from: https://gco.iarc.fr/. Access date: 06.12.2024.
2. Papamichael D, Audisio RA, Glimelius B, et al.. Treatment of colorectal cancer in older patients: International Society of Geriatric Oncology (SIOG) consensus recommendations 2013. Ann. Oncol., 2015;26(3):463-476. https://doi.org/10.1093/annonc/mdu253
3. Hossain MS, Karuniawati H, Jairoun AA, et al. (2022). Colorectal cancer: a review of carcinogenesis, global epidemiology, current challenges, risk factors, preventive and treatment strategies. Cancers, 2022;14(7):1732. https://doi.org/10.3390/cancers14071732
4. Testa U, Pelosi E, Castelli G. Colorectal cancer: genetic abnormalities, tumor progression, tumor heterogeneity, clonal evolution and tumor-initiating cells. Medical Sciences, 2018;6(2):31. https://doi.org/10.3390/medsci6020031
5. Baldwin LM., Dobie, SA, Billingsley K, et al. Explaining black–white differences in receipt of recommended colon cancer treatment. J Natl Cancer Inst, 2005;97(16):1211-1220. https://doi.org/10.1093/jnci/dji241
6. Abraha AM, Ketema EB. Apoptotic pathways as a therapeutic target for colorectal cancer treatment. World J Gastrointest Oncol, 2016;8(8):583. https://doi.org/10.4251/wjgo.v8.i8.583
7. Kiraz Y, Adan A, Kartal Yandim M, Baran Y. Major apoptotic mechanisms and genes involved in apoptosis. Tumour Biol, (2016);37:8471-8486. https://doi.org/10.1007/s13277-016-5035-9
8. Hossain MS, Urbi Z, Sule A, Rahman KH. Andrographis paniculata (Burm. f.) Wall. ex Nees: a review of ethnobotany, phytochemistry, and pharmacology. Sci. World, 2014: (1):274905. https://doi.org/10.1155/2014/274905
9. Urbi Z, Zainuddin Z. Standardization of surface sterilization protocol of field grown Stevia rebaudiana prior to in vitro clonal propagation. Jurnal Teknologi, 2015;77(24). https://doi.org/10.11113/jt.v77.6722
10. Hossain MS, Urbi Z. Effect of naphthalene acetic acid on the adventitious rooting in shoot cuttings of Andrographis paniculata (Burm. f.) Wall. ex Nees: an important therapeutical herb. Int. J. Agron, 2016;(1):1617543. https://doi.org/10.1155/2016/1617543
11. Vuković S, Popović-Djordjević JB, Kostić AŽ, et al. Allium species in the Balkan region—Major metabolites, antioxidant and antimicrobial properties. Horticulturae, 2023;9(3):408. https://doi.org/10.3390/horticulturae9030408
12. Iwar K, Ochar K, Seo YA, Ha BK., Kim SH. Alliums as Potential Antioxidants and Anticancer Agents. Int. J. Mol. Sci., 2024;25(15):8079. https://doi.org/10.3390/ijms25158079
13. Mohammadi-Motlagh HR, Mostafaie A, Mansouri K. Anticancer and anti-inflammatory activities of shallot (Allium ascalonicum) extract. Archives of Medical Science, 2011;7(1):38-44. https://doi.org/10.5114/aoms.2011.20602
14. Poonthananiwatkul B, Lim RH, Howard RL, Pibanpaknitee P, Williamson EM. Traditional medicine use by cancer patients in Thailand. J. Ethnopharmacol., 2015;168:100-107. https://doi.org/10.1016/j.jep.2015.03.057
15. Alkan M, Güzel M, Akşit H, Bağdat RB, Alkan FR, Evlice E. Chemical components and insecticidal effects of essential oils from three lavender cultivars against adult Sitophilus granarius (L., 1758)(Coleoptera: Curculionidae). Turk. j. entomol, 2021;45(4):405-416. https://doi.org/10.16970/entoted.988985
16. Wang YJ, Huang Y, Anreddy N. Tea nanoparticle, a safe and biocompatible nanocarrier, greatly potentiates the anticancer activity of doxorubicin. Oncotarget, 2015;7(5):5877. https://doi.org/10.18632/oncotarget.6711
17. Chandrashekar DS, Bashel B, Balasubramanya SAH, et al. UALCAN: a portal for facilitating tumor subgroup gene expression and survival analyses. Neoplasia, 2017;19(8):649-658. https://doi.org/10.1016/j.neo.2017.05.002
18. Men C, Chai H, Song X, Li Y, Du H, Ren Q. Identification of DNA methylation associated gene signatures in endometrial cancer via integrated analysis of DNA methylation and gene expression systematically. J. Gynecol. Oncol., 2017;28(6). https://doi.org/10.3802/jgo.2017.28.e83
19. Shinawi T, Hill VK, Krex D, et al. DNA methylation profiles of long-and short-term glioblastoma survivors. Epigenetics, 2013;8(2):149-156. https://doi.org/10.4161/epi.23398
20. Nagy Á, Győrffy B. muTarget: a platform linking gene expression changes and mutation status in solid tumors. Int J Cancer, 2021;148(2):502-511. https://doi.org/10.1002/ijc.33283
21. Menyhárt O, Harami-Papp H, Sukumar S, et al. Guidelines for the selection of functional assays to evaluate the hallmarks of cancer. Biochim. Biophys. Acta, Rev. Cancer, 2016;1866(2):300-319. https://doi.org/10.1016/j.bbcan.2016.10.002
22. Roshandel G, Ghasemi-Kebria F, Malekzadeh R. Colorectal Cancer: Epidemiology, Risk Factors, and Prevention. Cancers, 2024;16(8):1530. https://doi.org/10.3390/cancers16081530
23. Adebayo AS, Agbaje K, Adesina SK, Olajubutu O. Colorectal cancer: disease process, current treatment options, and future perspectives. Pharmaceutics, 2023;15(11):2620. https://doi.org/10.3390/pharmaceutics15112620
24. Wang Q, Shen X, Chen G, Du J. Drug resistance in colorectal cancer: from mechanism to clinic. Cancers, 2022;14(12):2928. https://doi.org/10.3390/cancers14122928
25. Kenmotsu H, Tanigawara Y. Pharmacokinetics, dynamics and toxicity of docetaxel: Why the Japanese dose differs from the Western dose. Cancer science, 2015;106(5):497-504. https://doi.org/10.1111/cas.12647
26. Beretta HV, Bannoud F, Insani M, et al. Relationships between bioactive compound content and the antiplatelet and antioxidant activities of six Allium vegetable species. Food Sci Biotechnol, 2017;55(2):266. https://doi.org/10.17113/ftb.55.02.17.4722
27. Suleria HAR, Butt MS, Anjum FM, Saeed F, Khalid N. Onion: Nature protection against physiological threats. Crit. Rev. Food Sci. Nutr., 2015;55(1):50-66. https://doi.org/10.1080/10408398.2011.646364
28. Sharifi-Rad J, Mnayer D, Tabanelli G, et al. Plants of the genus Allium as antibacterial agents: From tradition to pharmacy. Cell. Mol. Biol., 2016;62(9):57-68. https://doi.org/10.14715/cmb/2016.62.9.10
29. Abdelrahman M, Mahmoud HY, El-Sayed M, Tanaka S, Tran LS. Isolation and characterization of Cepa2, a natural alliospiroside A, from shallot (Allium cepa L. Aggregatum group) with anticancer activity. Plant Physiol Biochem, 2017;116:167-173. https://doi.org/10.1016/j.plaphy.2017.05.006
30. Kazemi M, Zolfaghari B, Keyvanlo Shahrestanaki M, Sadeghi Dinani M. Cytotoxic Effects of Allium affine Ledeb Butanolic Fraction on Breast and Ovary Cancer Cell Lines. J. Med. Plants 2017;16(64):83-90.
31. Abdel-Hady H, El-Sayed MM, Abdel-Gawad MM, El-Wakil EA, Abdel-Hameed ESS, Abdel-Lateef EES. LC-ESI-MS analysis, antitumor and antioxidant activities of methanolic extract of Egyptian Allium kurrat. J. Appl. Pharm. Sci., 2018;8(7):085-092. https://doi.org/10.7324/JAPS.2018.8714
32. Khazaei S, Esa NM, Ramachandran V, Hamid RA, Pandurangan AK, Etemad A, Ismail P. In vitro antiproliferative and apoptosis inducing effect of Allium atroviolaceum bulb extract on breast, cervical, and liver cancer cells. Front. pharmacol., 2017;8: 5. https://doi.org/10.3389/fphar.2017.00005
33. Khazaei S, Abdul Hamid R, Mohd Esa N, Ramachandran V, Aalam GTF, Etemad A, Ismail P. Promotion of HepG2 cell apoptosis by flower of Allium atroviolaceum and the mechanism of action. BMC Complement Altern Med, 2017;17:1-13. https://doi.org/10.1186/s12906-017-1594-6
34. Ivanova A, Mikhova B, Najdenski H, Tsvetkova I, Kostova I. Chemical composition and antimicrobial activity of wild garlic Allium ursinum of Bulgarian origin. Nat. Prod. Commun., 2009;4(8):1934578X0900400808. https://doi.org/10.1177/1934578X0900400808
35. Xu XY, Song GQ, Yu YQ, Ma HY, Ma L, Jin YN. Apoptosis and G2/M arrest induced by Allium ursinum (ramson) watery extract in an AGS gastric cancer cell line. Onco Targets Ther, 2013;779-783. https://doi.org/10.2147/OTT.S45865
36. Pârvu AE, Cătoi F, Deelawar S, Sarup D, Pârvu M. Anti-inflammatory effect of Allium ursinum. Not Sci Biol, 2014;6(1):20-26. https://doi.org/10.15835/nsb619252
37. Stanisavljević N, Soković Bajić S, Jovanović Ž, et al. Antioxidant and antiproliferative activity of Allium ursinum and their associated microbiota during simulated in vitro digestion in the presence of food matrix. Front. microbiol., 2020;11:601616. https://doi.org/10.3389/fmicb.2020.601616
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Toplam 47 adet kaynakça vardır.

Ayrıntılar

Birincil Dil	İngilizce
Konular	Tıp Eğitimi
Bölüm	Araştırma Makalesi
Yazarlar	Zuhal Tunçbilek 0000-0002-6510-0884 Gonca Kabak 0009-0004-2430-5076 Yavuz Siliğ 0000-0002-0562-7457
Yayımlanma Tarihi	31 Aralık 2024
Gönderilme Tarihi	13 Aralık 2024
Kabul Tarihi	25 Aralık 2024
Yayımlandığı Sayı	Yıl 2024Cilt: 46 Sayı: 4

Kaynak Göster

AMA	Tunçbilek Z, Kabak G, Siliğ Y. Investigation of the Synergistic Effect of Allium Polyanthum Schult Extract and Docetaxel on Apoptosis-Related Genes in Colorectal Cancer Cells and Bioinformatics Analysis. CMJ. Aralık 2024;46(4):280-291. doi:10.7197/cmj.1601162

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