Dianthus orientalisin pankreas kanserindeki invazyon ve metastaz etkilerinin in vitro olarak araştırılması
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Tarih
2024
Yazarlar
Dergi Başlığı
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Cilt Başlığı
Yayıncı
Necmettin Erbakan Üniversitesi, Tıp Fakültesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Amaç: Pankreas kanseri ileri derece ölümcül bir malign hastalıktır. Pankreas kanseri
tedavisinin hastalık evresine bağlı olarak cerrahi teknikler mevcut olup beraberinde birden
fazla sistemik tedavi seçenekleri mevcuttur (Mizrahi ve ark. 2020). Hastaların tedavisinde
kullanılan sistemik kemoterapötikler kanser hücrelerinin apopitozunu sağlayabilir,
proliferasyonunu engelleyebilir, metastatik özelliklerini baskılayabilir veya hücre ölüm
yolaklarını uyarararak etki gösterebilmektedir. Pankreas kanserinin tedavisinde hem sentetik
hem de bitkisel kaynaklı ilaç araştırmaları halen devam etmektedir. Bununla birlikte
neoadjuvan kemoterapide ilaç toksisite veya perioperatif komplikasyonların artışına sebep
olma gibi dezavantajları mevcuttur. Pankreas kanserinde adjuvan ve neoadjuvan yeni
ajanların çıkması hastalara sunulacak tedaviler için ileri derece önem arz etmektedir. Klinik
kullanımda olan ilaçların çoğunluğu bitkisel kaynaklı olup dianthus türleri de içerdiği fenolik
bileşikler ile antikanserojen bir ajan olabilecek potansiyele sahiptir. Bununla birlikte
literatürde bu bitki ile yapılmış az sayıda veri bulunmaktadır. Bu çalışmada Dianthus
orientalis ekstraktının pankreas kanser hücre hattındaki olası antikanser, antiproliferatif ve
antimetastatik etkilerinin değerlendirilmesi amaçlandı.
Materyal ve Metod: Çalışmada Panc-1 ve BxPC-3 olmak üzere iki farklı pankreas hücre
hattı kullandı. Dianthus orientelis ekstraklarının sağlıklı hücrelerdeki etkisinin
değerlendirilmesi amacıyla HEK-293 sağlıklı embriyonik böbrek hücre hattı kullanıldı.
Çalışma için Dianthus orientalis ekstraklarının uygulanıp uygulanmadığı duruma göre;
Panc-1 hücre hattına Dianthus orientalis’in uygulandığı grup DiO-[P+], uygulanmadığı
grup DiO-[Pneg], BxPC-3 hücre hattına Dianthus orientalis’in uygulandığı grup DiO-
[Bx+], uygulanmadığı grup DiO-[Bxneg] HEK293 hücrelerine DiO ekstraklarının
uygulandığı grup DiO-[HEK+] ve uygulanmadığı grup DiO-[HEKneg] olmak üzere gruplar
oluşturuldu. Panc-1, BxPC-3 ve HEK-293 hücreleri %10 FBS (Fetal Bovine Serum) ve %1
penisilin/streptomisin (100 μg/ml streptomisin ve 100 U/ml penisilin) içeren DMEM
(Dulbecco's Modified Eagle's Mediu) kültürü ortamında T75 flasklarda 37°C ve % 5’lik
CO2’li ortamda kültüre edildi. Hücrelerin bu ekstrakta duyarlılığı tetrazolyum indirgenme
(MTT) methoduyla analiz edildi. RT-PZR yöntemi ile Bax, Bad, p50, Bak, Apaf-1, Bcl-2,
Bcl-XL, Casp6, Casp9, Casp3, Casp12, Casp2, p50, Bak, Apaf-1, MMP7, MMP9, VIM,
SNAIL, N-Cadherin, Beta-Catenin, ZEB1, ACTA2, DDR2, SNAI2, SNAI1, PALB2, CHEK2,
CDH1, TWIST1 gen ekspresyonları analiz edildi.
Bulgular: Çalışmamızda Panc-1 ve Bx-PC3 Pankreas kanseri hücre hattında Dianthus
orientalis ekstraktı ile tedavi edilen grupta SNAI1 PALB2 ve CHEK2 ekpsresyonları anlamlı
olarak up-regüle idi. Bx-PC3 hücre hattında CHD1 ekspresyonu anlamlı olarak down-regüle
idi . Apopitoz analizinde tedavi grubunda kontrol grubuna göre artmış apopitoz oranı
görüldü.
Sonuçlar: MTT analiz sonucunda göre IC50 dozu 48 saatte 250 μg olarak tespit edildi.
Çalışmamız pankreas hücre hattında uygulanılan Dianthus orientalis ekstraktı için invazyon
ve metataz üzerine bazı genlerde anlamlı farklılıklar tespit edildi. Proapototik genler ile
kaspaz yolağındaki başlatıcı ve efektör kaspaz aktivasyonu sağlayarak apopitoz yolağını
aktive ettiğini gösterdi, apopitoz analizindeki kontrole göre artmış apopitoz oranı ekspresyon
sonuçlarını da destekler nitelikteydi. Sonuç olarak pankreas kanseri hücresine Dianthus
orientalis ektraktının anlamlı antikansoreojen ekinliği gösterilmiştir.
Aim: Pancreatic cancer is a highly lethal malignant disease. Treatment of pancreatic cancer has multiple systemic treatment options, including surgical techniques, depending on the stage of the disease (Mizrahi ve ark. 2020).Systemic chemotherapeutic treatments used in the treatment of patients can cause the apoptosis of cancer cells, prevent their proliferation, suppress their metastatic properties, or act by stimulating cell death pathways. However, neoadjuvant chemotherapy has dis advantages such as toxicity or increased perioperative complications. The emergence of new adjuvant and neoadjuvant agents in pancreatic cancer is of great importance for the treatments offered to patients.The majority of drugs in clinical use are of plant origin, and Dianthus species also have the potential to be anticancer agents with the phenolic compounds they contain. However, there are few data on this plant in the literature. Therefore, the study aimed to evaluate the possible anticancer, antiproliferative and antimetastatic effects of Dianthus orientalis extract on the pancreatic cancer cell line. Material and Method: Two different pancreatic cell lines were used in the study: Panc-1 and BxPC-3. HEK-293 healthy embryonic kidney cell line was used to evaluate the effect of Dianthus orientalis extracts on healthy cells. Depending on whether Dianthus orientalis extracts are applied for the study or not; The group in which Dianthus orientalis was applied to the Panc-1 cell line was DiO-[P+], the group in which it was not applied was DiO-[Pneg], the group in which Dianthus orientalis was applied to the BxPC-3 cell line was DiO-[Bx+], the group in which it was not applied was DiO-[Bxneg] HEK293 Groups were created: DiO- [HEK+], the group in which DiO extracts were applied to the cells, and DiO-[HEKneg], the group in which DiO extracts were not applied. Panc-1, BxPC-3 and HEK-293 cells are T75 in DMEM (Dulbecco's Modified Eagle's Mediu) culture medium containing 10% FBS (Fetal Bovine Serum) and 1% penicillin/streptomycin (100 μg/ml streptomycin and 100 U/ml penicillin). It was cultured in flasks at 37°C and 5% CO2. The sensitivity of cells to this extract was analyzed by the tetrazolium reduction (MTT) method. Bax, Bad, p50, Bak, Apaf- 1, Bcl-2, Bcl-XL, Casp6, Casp9, Casp3, Casp12, Casp2, p50, Bak, Apaf-1, MMP7, MMP9, VIM, SNAIL with RT-PCR method , N-Cadherin, Beta-Catenin, ZEB1, ACTA2, DDR2, SNAI2, SNAI1, PALB2, CHEK2, CDH1, TWIST1 gene expressions were analyzed. Findings: In our study, SNAI1, PALB2 and CHEK2 expressions were significantly upregulated in the group treated with Dianthus orientalis extract in the Panc-1 and Bx-PC3 Pancreatic cancer cell line. In the apoptosis analysis, an increased rate of apoptosis was observed in the treatment group compared to the control group. Results: According to the MTT analysis result, the IC50 dose was determined as 250 μg in 48 hours. In our study, significant differences were detected in some genes on invasion and metatasis for the Dianthus orientalis extract applied to the pancreas cell line. It showed that it activated the apoptosis pathway by activating pro-apototic genes and the initiator and effector caspase in the caspase pathway. The increased rate of apoptosis compared to the control in the apoptosis analysis also supported the expression results. As a result, significant anticarcinogenic activity of Dianthus orientalis extract against pancreatic cancer cells was demonstrated.
Aim: Pancreatic cancer is a highly lethal malignant disease. Treatment of pancreatic cancer has multiple systemic treatment options, including surgical techniques, depending on the stage of the disease (Mizrahi ve ark. 2020).Systemic chemotherapeutic treatments used in the treatment of patients can cause the apoptosis of cancer cells, prevent their proliferation, suppress their metastatic properties, or act by stimulating cell death pathways. However, neoadjuvant chemotherapy has dis advantages such as toxicity or increased perioperative complications. The emergence of new adjuvant and neoadjuvant agents in pancreatic cancer is of great importance for the treatments offered to patients.The majority of drugs in clinical use are of plant origin, and Dianthus species also have the potential to be anticancer agents with the phenolic compounds they contain. However, there are few data on this plant in the literature. Therefore, the study aimed to evaluate the possible anticancer, antiproliferative and antimetastatic effects of Dianthus orientalis extract on the pancreatic cancer cell line. Material and Method: Two different pancreatic cell lines were used in the study: Panc-1 and BxPC-3. HEK-293 healthy embryonic kidney cell line was used to evaluate the effect of Dianthus orientalis extracts on healthy cells. Depending on whether Dianthus orientalis extracts are applied for the study or not; The group in which Dianthus orientalis was applied to the Panc-1 cell line was DiO-[P+], the group in which it was not applied was DiO-[Pneg], the group in which Dianthus orientalis was applied to the BxPC-3 cell line was DiO-[Bx+], the group in which it was not applied was DiO-[Bxneg] HEK293 Groups were created: DiO- [HEK+], the group in which DiO extracts were applied to the cells, and DiO-[HEKneg], the group in which DiO extracts were not applied. Panc-1, BxPC-3 and HEK-293 cells are T75 in DMEM (Dulbecco's Modified Eagle's Mediu) culture medium containing 10% FBS (Fetal Bovine Serum) and 1% penicillin/streptomycin (100 μg/ml streptomycin and 100 U/ml penicillin). It was cultured in flasks at 37°C and 5% CO2. The sensitivity of cells to this extract was analyzed by the tetrazolium reduction (MTT) method. Bax, Bad, p50, Bak, Apaf- 1, Bcl-2, Bcl-XL, Casp6, Casp9, Casp3, Casp12, Casp2, p50, Bak, Apaf-1, MMP7, MMP9, VIM, SNAIL with RT-PCR method , N-Cadherin, Beta-Catenin, ZEB1, ACTA2, DDR2, SNAI2, SNAI1, PALB2, CHEK2, CDH1, TWIST1 gene expressions were analyzed. Findings: In our study, SNAI1, PALB2 and CHEK2 expressions were significantly upregulated in the group treated with Dianthus orientalis extract in the Panc-1 and Bx-PC3 Pancreatic cancer cell line. In the apoptosis analysis, an increased rate of apoptosis was observed in the treatment group compared to the control group. Results: According to the MTT analysis result, the IC50 dose was determined as 250 μg in 48 hours. In our study, significant differences were detected in some genes on invasion and metatasis for the Dianthus orientalis extract applied to the pancreas cell line. It showed that it activated the apoptosis pathway by activating pro-apototic genes and the initiator and effector caspase in the caspase pathway. The increased rate of apoptosis compared to the control in the apoptosis analysis also supported the expression results. As a result, significant anticarcinogenic activity of Dianthus orientalis extract against pancreatic cancer cells was demonstrated.
Açıklama
Anahtar Kelimeler
Pankreas, Pancreas, Apopitoz, Apoptosis, Metastaz, Metastasis, Dianthus orientalis, Antikanserojen, Anticancer
Kaynak
WoS Q Değeri
Scopus Q Değeri
Cilt
Sayı
Künye
Sevinç, B. (2024). Dianthus orientalisin pankreas kanserindeki invazyon ve metastaz etkilerinin in vitro olarak araştırılması. (Yayınlanmamış tıpta uzmanlık tezi) Necmettin Erbakan Üniversitesi, Tıp Fakültesi Cerrahi Tıp Bilimleri Bölümü Genel Cerrahi Anabilim Dalı, Konya.
