Neferin molekülünün prostat kanser hücrelerinin kemoterapi duyarlılığına etkisi
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Dosyalar
Tarih
2019
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Trakya Üniversitesi Sosyal Bilimler Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Prostat kanseri dünya genelinde erkeklerin en ölümcül hastalıklarından biridir. Bu çalışmanın amacı, geleneksel bir tıbbi bitki olan Nelumbo nucifera'nın (Lotus) tohum embriyosundan izole edilen neferin, bisbenzilizokinolin alkaloidin insan prostat kanser hücrelerinde doksorubisin tedavisi üzerindeki etkisini araştırmaktı. Prostat kanseri hücreleri, neferin, doksorubisin veya ikisinin kombinasyonu ile tedavi edildi. Hücre sağkalımı MTT testi ile belirlendi. Hücresel apoptoz ve hücre döngü evreleri sitometre cihazında değerlendirildi. Hücre migrasyonu yara iyileşme testi ile incelendi. RT-qPCR, mRNA ekspresyonunu değerlendirmek için kullanıldı. Androjen-dirençli PC3 ve androjen-duyarlı LNCaP hücrelerinin neferin ile tedavisi, doz ve zamana bağlı bir şekilde hücre canlılığını önemli ölçüde azalttı. Bir kemoterapötik ajan olan doksorubisinin neferin ile kombinasyonu, anti-proliferatif aktiviteyi anlamlı düzeyde artırdı. Sitometrik değerlendirmeye göre neferin doksorubisinin apoptoz ve G1 fazında hücre siklusunu durması üzerindeki etkilerini uyarmaktadır. PCR analizleri, kombine tedavi rejiminin kaspaz-3, -8, sitokrom c, p53'ü indükleyerek ve Bcl-XL ve survivin mRNA ekspresyonunu azaltarak apoptoza yol açtığını gösterdi. Neferin tedavisi Beklin-1 ve ATG7 genlerini indükleyerek otofajiye neden olmaktadır. Hücre döngüsü, G1 fazında artmış p21 ve düşürülmüş CDK2, CDK4 ve CDK6 regülasyonu ile duraklatıldı. Ayrıca, neferin hücre migrasyonunu inhibe ederek doksorubisin tedavisine anti-migrasyon yoluyla da katkı sağladı. Bu çalışmadan elde edilen verilere göre, neferin androjen-dirençli ve androjen-duyarlı prostat kanseri tedavisinde tek ya da kombine uygulanabilme potansiyeline sahip olabileceği, ancak ileri in vivo araştırmalar ile desteklenmesi gerektiği sonucuna varıldı.
Prostate cancer (PCa) is one of the most lethal diseases of men worldwide. The aim of this study was to investigate the effect of neferine, bisbenzylisoquinoline alkaloid isolated from the seed embryo of a traditional medicinal plant Nelumbo nucifera (Lotus) on doxorubicin therapy in human PCa cells. Prostate cancer cells were treated with neferine with or without doxorubicin. Cell survival was tested by MTT assay. Cell apoptosis were assessed by annexin V staining with cytometry. Cell cycle progression was determined with cell-based cytometer. Cell migration was evaluated by wound healing test. RT-qPCR was used to assess mRNA expression. Neferine treatment of androgen-insensitive PC3 and androgen-sensitive LNCaP cells significantly decreased cell viability in a dose and time-dependent manner. Combination of neferine and doxorubicin, a chemotherapeutic agent, significantly enhanced its anti-proliferative activity. Cytometric evaluation demonstrated that neferine induces effects of doxorubicin on apoptosis and cell cycle arrest at G1 phase. PCR analyzes demonstrated that therapy regimes lead apoptosis through inducing caspase 3 and 8, Bax, p53, and decreased the mRNA expression of Bcl-XL. Cell cycles were arrested at G1 phase by upregulated p21 and down regulation of cyclin dependent kinases (CDK) such as CDK2, CDK4 and CDK6. In addition, neferine contributed to doxorubicin treatment by anti-migration via inhibiting cell migration. According to the data obtained in this study, it was concluded that neferine may have single or combined potency in the treatment of androgen-resistant and androgen-sensitive prostate cancer but should be supported by further in vivo studies.
Prostate cancer (PCa) is one of the most lethal diseases of men worldwide. The aim of this study was to investigate the effect of neferine, bisbenzylisoquinoline alkaloid isolated from the seed embryo of a traditional medicinal plant Nelumbo nucifera (Lotus) on doxorubicin therapy in human PCa cells. Prostate cancer cells were treated with neferine with or without doxorubicin. Cell survival was tested by MTT assay. Cell apoptosis were assessed by annexin V staining with cytometry. Cell cycle progression was determined with cell-based cytometer. Cell migration was evaluated by wound healing test. RT-qPCR was used to assess mRNA expression. Neferine treatment of androgen-insensitive PC3 and androgen-sensitive LNCaP cells significantly decreased cell viability in a dose and time-dependent manner. Combination of neferine and doxorubicin, a chemotherapeutic agent, significantly enhanced its anti-proliferative activity. Cytometric evaluation demonstrated that neferine induces effects of doxorubicin on apoptosis and cell cycle arrest at G1 phase. PCR analyzes demonstrated that therapy regimes lead apoptosis through inducing caspase 3 and 8, Bax, p53, and decreased the mRNA expression of Bcl-XL. Cell cycles were arrested at G1 phase by upregulated p21 and down regulation of cyclin dependent kinases (CDK) such as CDK2, CDK4 and CDK6. In addition, neferine contributed to doxorubicin treatment by anti-migration via inhibiting cell migration. According to the data obtained in this study, it was concluded that neferine may have single or combined potency in the treatment of androgen-resistant and androgen-sensitive prostate cancer but should be supported by further in vivo studies.
Açıklama
Anahtar Kelimeler
Prostat Kanseri, Neferin, Doksorubisin, PC3, LNCaP, Apoptoz, Prostate Cancer, Neferine, Doxorubicin, PC3, LNCaP, Apoptosis
