VİTİS VİNİFERA ÇEKİRDEK ÖZÜTÜ KULLANILARAK ALTIN NANOPARTİKÜLLERİN YEŞİL SENTEZİ VE KOLON KANSERİ (HT-29) HÜCRELERİNDE ANTİ- KANSER ÖZELLİKLERİNİN DEĞERLENDİRİLMESİ

Yıl 2021, Cilt: 28 Sayı: 3, 455 - 464, 13.09.2021

Selim Genç Suray Pehlivanoğlu Çiğdem Aydın Acar Şükriye Yeşilot

https://doi.org/10.17343/sdutfd.875236

Öz

Amaç
Son yıllarda metalik nanopartiküller antioksidan, antimikrobiyal
ve antikanser etkinlikleri ile nanotıp alanında
oldukça dikkat çekmektedir. Özellikle, kanser
tedavisinde farmakolojik etkinliği bilinen tıbbi bitkilerden
yeşil sentez metodu ile üretilen metal nanopartiküller
araştırmacıların ilgi odağı haline gelmiştir. Altın
nanopartiküller; kendilerine özgü kimyasal, fiziksel ve
nontoksik özellikleri ile ön plana çıkan metal nanopartiküllerden
biridir. Çalışmamızda, üzüm (Vitis vinifera)
çekirdeği sulu ekstraktı kullanılarak yeşil sentezi
gerçekleştirilen altın nanopartiküllerin (Vv-AuNP) tek
başına ve kemoterapötik bir ajan olan Gemsitabin ile
kombine kullanımının HT-29 kolon kanseri hücre hattı
üzerindeki sitotoksik, anti-proliferatif ve apoptotik etkilerinin
değerlendirilmesi amaçlanmıştır.
Gereç ve Yöntem
Altın nanopartiküllerin yeşil sentezi, Vitis vinifera
sulu özütü kullanılarak gerçekleştirildi. Vv-AuNP’lerin
UV-görünür spektrofotometre, taramalı elektron
mikroskobu (SEM) ve Enerji Dağılımlı X-ışını Spektroskopisi
(EDX) gibi çeşitli analitik tekniklerle karakterizasyonu
yapıldı. Vv-AuNP'ler 0–80 μg/mL doz
aralığında tek başına ve Gemsitabin (100 μg/mL veya
200 μg/mL sabit doz) ile kombine olacak şekilde HT-
29 hücreleri ile 72 saat süresince muamele edildi.
Sitotoksik etkileri MTT testi ile değerlendirildi. Ayrıca,
HT-29 hücreleri üzerine Vv-AuNP’lerin artan dozlarda
(0-400μg/mL) tek başına ve Gemsitabin ile kombine
antiproliferatif etkileri ise BrdU testi ile belirlendi. Altın
nanopartiküllerin HT-29 hücreleri ile muamelesi sonrası
Kaspaz-3 aktivasyon düzeyi Western-blot yöntemi
ile analiz edildi.
Bulgular
MTT sonuçları değerlendirildiğinde Vv-AuNP’lerin 80
μg/mL doza kadar çıkıldığında HT-29 hücrelerinde
istenilen düzeyde sitotoksik etkiler göstermediği belirlendi.
BrdU hücre proliferasyon testi sonuçlarına dayalı
olarak, Vv-AuNP’lerin tek başına ve Gemsitabin
ile birlikte uygulamalarında IC50 değerleri sırasıyla
147.9 ve 39.43 μg/mL olarak belirlendi. Vv-AuNP dozlarına
bağlı kaspaz-3 aktivasyonunun kontrole kıyasla
arttığı, Gemsitabin ile kombine kullanımının bu etkiyi
güçlendirdiği gözlendi.
Sonuç
Sonuç olarak, yeşil sentezlenmiş AuNP’ler in vitro koşullarda
kolon kanseri hücreleri üzerinde anti-kanser
özellikler sergilemiştir. Bu çalışmanın sonuçları, Vv-AuNP'lerin
kolon kanseri tedavisi için potansiyel bir seçenek
olarak düşünülebileceğini göstermektedir.

Anahtar Kelimeler

Vitis vinifera, Altın Nanopartikül, Gemsitabin, Kolon Kanseri, HT-29

Destekleyen Kurum

Burudur Mehmet Akif Ersoy Üniversitesi Bilimsel Araştırma Projeleri Koordinatörlüğü

Proje Numarası

0595-YL-19

GREEN SYNTHESIS OF GOLD NANOPARTICLES USING VITIS VINIFERA SEED EXTRACT AND EVALUATION OF ANTI-CANCER PROPERTIES IN COLON CANCER (HT-29) CELLS

Öz

Objective
In recent years, metallic nanoparticles have attracted
the attention in the field of nanomedicine with their
antioxidant, antimicrobial and anticancer activities.
Especially, metal nanoparticles produced by green
synthesis method from medicinal plants with known
pharmacological efficacy in cancer treatment have
been spotlighted by researchers.. Gold nanoparticles;
become prominentamong metal nanoparticles
with their unique chemical, physical and non-toxic
properties. In this study, it was aimed to perform
green synthesis of gold nanoparticles using Vitis
vinifera (grape seed) aqueous extract and to evaluate
cytotoxic, anti-proliferative and apoptotic effects
of gold nanoparticles (Vv-AuNP) alone and with
Gemcitabine co-administration on HT-29 human
colon cancer cell line.
Materials and Methods
Green synthesis of gold nanoparticles was carried out
using the aqueous extract of Vitis vinifera. Vv-AuNPs
were characterized by various analytical techniques
such as UV-visible spectrophotometer, scanning
electron microscope (SEM) and Energy Dispersive
X-ray Spectroscopy (EDX). Vv-AuNPs were treated
with HT-29 cells at a dose range of 0–80 μg / mL for
72 hours, alone and in combination with Gemcitabine
(100 μg / mL or 200 μg / mL fixed dose). Cytotoxic
effects were evaluated by MTT assay. In addition,
the antiproliferative effects of Vv-AuNPs at increasing
doses (0-400 μg / mL) alone and combined with
Gemcitabine were determined by the BrdU test. After
treatment of gold nanoparticles with HT-29 cells,
the level of Caspase-3 activation was analyzed by
Western-blot method.
Results
According to MTT results, when the dose of Vv-
AuNPs was increased to 80 μg / mL, it was determined
that HT-29 cells did not show the expected level of
cytotoxic effect. According to the results of the BrdU
cell proliferation test, IC50 values of Vv-AuNPs alone
and combined with Gemcitabine were determined
as 147.9 and 39.43 μg / mL, respectively. It was
observed that the activation of caspase-3 due to Vv-
AuNP doses increased compared to control, and its
combined use with Gemcitabine reinforced this effect.
Conclusion
As a result, green synthesized AuNPs exhibited
anti-cancer properties on colon cancer cells in vitro.
The results of this study show that Vv-AuNPs can
be considered as a potential option for colon cancer
treatment.

Anahtar Kelimeler

Vitis vinifera, Gold Nanoparticle, Gemcitabine, Colon Cancer, HT-29

Proje Numarası

0595-YL-19

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