Protoporfirin IX ve Sono-Fotodinamik Terapi ile HepG2 Karaciğer Kanseri Hücrelerinde Apoptozun Sinerjistik Artışı

Öz

Hepatoselüler karsinom (HSK), ileri evrelerde etkili tedavilerden yoksundur. Bu çalışma, Protoporfirin IX (PpIX) aracılı sonodinamik (SDT), fotodinamik (PDT) ve kombine sono-fotodinamik terapinin (SPDT) HepG2 hücreleri üzerindeki sitotoksik ve pro-apoptotik etkilerini değerlendirmeyi amaçlamıştır. HepG2 hücreleri, 50 µM PpIX ile tek başına veya SDT (1 MHz, 0.5 W/cm², 60 s), PDT (630 nm, 2 mJ/cm², 30 s) ya da ardışık SPDT ile kombine edilerek muamele edildi. Canlılık (MTT), oksidatif stres (TOC, TAC), apoptoz (Annexin V-FITC/PI boyaması) ve apoptotik protein ekspresyonu (Western blot ile Bax, Bcl-2, kesilmiş kaspaz-3, -9) değerlendirildi. Tek başına PpIX sitotoksisite göstermedi (p > 0,05). SDT ve PDT, canlılığı sırasıyla %24,33 (p < 0,05) ve %43,33 (p < 0,01) oranında anlamlı olarak azaltırken, SPDT canlılığı %83,33 (p < 0,001) oranında azalttı. TOC seviyeleri SDT ve PDT gruplarında anlamlı şekilde arttı (p < 0,01) ve SPDT grubunda en yüksek seviyeye ulaştı (p < 0,001). Apoptoz oranları en yüksek SPDT ile elde edildi (tüm gruplara karşı p < 0,001). SPDT, Bax ve kesilmiş kaspazların ekspresyonunu belirgin şekilde artırırken, Bcl-2'yi düşürdü (p < 0,001). PpIX aracılı SPDT, HepG2 hücrelerinde artırılmış oksidatif stres ve mitokondriyal apoptoz yolaklarının aktivasyonu yoluyla sinerjistik sitotoksisite indüklemektedir. Bu bulgular, SPDT'nin HSK tedavisi için umut verici bir strateji olduğunu ve daha ileri preklinik araştırmaları gerektirdiğini göstermektedir.

Anahtar Kelimeler

• Protoporfirin IX
• Hepatoselüler karsinom
• Sonodinamik tedavi
• Fotodinamik tedavi
• Apoptoz
• Oksidatif stres

Synergistic Enhancement of Apoptosis by Protoporphyrin IX-Mediated Sono-Photodynamic Therapy in HepG2 Hepatocellular Carcinoma Cells

Öz

Hepatocellular carcinoma (HCC) lacks effective therapies at advanced stages. This study aimed to evaluate the cytotoxic and pro-apoptotic effects of Protoporphyrin IX (PpIX)-mediated sonodynamic (SDT), photodynamic (PDT), and combined sono-photodynamic therapy (SPDT) on HepG2 cells. HepG2 cells were treated with 50 µM PpIX alone or combined with SDT (1 MHz, 0.5 W/cm², 60 s), PDT (630 nm, 2 mJ/cm², 30 s), or sequential SPDT. Viability (MTT), oxidative stress (TOC, TAC), apoptosis (Annexin V-FITC/PI staining), and apoptotic protein expression (Bax, Bcl-2, cleaved caspase-3, -9 via Western blot) were assessed.PpIX alone showed no cytotoxicity (p > 0.05). SDT and PDT significantly reduced viability by 24.33% (p < 0.05) and 43.33% (p < 0.01), respectively, while SPDT reduced it by 83.33% (p < 0.001). TOC levels increased significantly in SDT and PDT groups (p < 0.01), peaking in SPDT (p < 0.001). Apoptosis rates were highest with SPDT (p < 0.001 vs. all). SPDT markedly upregulated Bax and cleaved caspases while downregulating Bcl-2 (p < 0.001).PpIX-mediated SPDT induces synergistic cytotoxicity in HepG2 cells through enhanced oxidative stress and activation of mitochondrial apoptosis pathways. These findings suggest SPDT as a promising strategy for HCC treatment, warranting further preclinical investigation.

Anahtar Kelimeler

• Protoporphyrin IX
• Hepatocellular carcinoma
• Sonodynamic therapy
• Photodynamic therapy
• Apoptosis
• Oxidative stress

Kaynakça

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