Institute of Sciences, Department of Biology, Kırıkkale University, Kırıkkale, Yahşihan, Turkey.
Vocational High School of Health Care Services, Department of Medical Services and Techniques, Kırıkkale University, Kırıkkale, Yahşihan, Turkey.
PeerJ. 2024 Nov 12;12:e18214. doi: 10.7717/peerj.18214. eCollection 2024.
Pancreatic carcinoma is an aggressive cancer that progresses without many symptoms. The difficulty of early diagnosis and an inadequate response to traditional treatments also cause the survival rate of pancreatic cancer to be low. Current research is focusing on methods of diagnosis and treatment, such as gene therapy, to increase survival rates. Small interfering ribonucleic acid (siRNA) has emerged as a promising advanced therapeutic strategy for cancer treatment. This study sought to silence the KRAS gene in the human pancreatic carcinoma cell line using a complex of small interfering ribonucleic acid (siRNA) and gold nanoparticles (AuNP).
In this study, 25 nM siRNA and gold nanoparticles at 0.5 mg/ml, 0.25 mg/ml, and 0.125 mg/ml concentrations were used to silence the KRAS gene in the CAPAN-1 cell line. Real-time PCR analysis, agarose gel electrophoresis, and double staining were carried out, and xCelligence real-time cell analysis (RTCA) was used to measure proliferation.
The PCR analysis revealed crossing point (CP) values of actin beta (ACTB) ranging from 33.04 to 35.98, which was in the expected range for all samples. The interaction between the gold nanoparticle/siRNA complex in the double staining analysis revealed that the most effective concentration of gold nanoparticle was 0.125 mg/ml. The WST-1 technique showed that siRNA/AuPEI cells in application groups had a viability rate of over 90%, indicating no toxicity or side effects. The xCELLigence RTCA® showed that at hour 72, there was a significant difference in proliferation in the 0.5 mg/mL PEI/AuNP-siRNA, 0.25 mg/mL PEI/AuNP-siRNA, and 0.125 mg/mL PEI/AuNP-siRNA application groups compared to the control and siRNA groups ( < 0.05). By hour 96, all three groups were statistically different from the control and siRNA groups in terms of proliferation ( < 0.05).
The results of this analysis suggest that the AuPEI/siRNA complex can be effectively used to silence the target gene, but more studies are needed to verify these results.
胰腺癌是一种侵袭性很强的癌症，在没有明显症状的情况下进展。早期诊断的困难和对传统治疗的反应不足也是导致胰腺癌存活率低的原因。目前的研究集中在诊断和治疗方法上，如基因治疗，以提高存活率。小干扰核糖核酸 (siRNA) 已成为癌症治疗的一种很有前途的先进治疗策略。本研究旨在使用小干扰核糖核酸 (siRNA) 和金纳米粒子 (AuNP) 复合物沉默人胰腺癌细胞系中的 KRAS 基因。
在这项研究中，使用浓度为 25 nM 的 siRNA 和浓度为 0.5 mg/ml、0.25 mg/ml 和 0.125 mg/ml 的金纳米粒子来沉默 CAPAN-1 细胞系中的 KRAS 基因。进行实时 PCR 分析、琼脂糖凝胶电泳和双重染色，并使用 xCelligence 实时细胞分析 (RTCA) 测量增殖。
PCR 分析显示肌动蛋白β (ACTB) 的 CP 值范围为 33.04 至 35.98，所有样本均在预期范围内。在双重染色分析中，金纳米粒子/siRNA 复合物的相互作用表明最有效的金纳米粒子浓度为 0.125 mg/ml。WST-1 技术表明，应用组中的 siRNA/AuPEI 细胞的存活率超过 90%，表明没有毒性或副作用。xCELLigence RTCA® 显示，在 72 小时时，0.5 mg/ml PEI/AuNP-siRNA、0.25 mg/ml PEI/AuNP-siRNA 和 0.125 mg/ml PEI/AuNP-siRNA 应用组与对照组和 siRNA 组相比，增殖有显著差异（<0.05）。在 96 小时时，所有三组与对照组和 siRNA 组在增殖方面均有统计学差异（<0.05）。
本分析结果表明，AuPEI/siRNA 复合物可有效用于沉默靶基因，但需要进一步研究来验证这些结果。
