Zaferani Arani Hamid, Abbasy Zahra, Abbasy Seyed Mahmoud Reza, Fooladivanda Nastaran, Kheradmand Mahdi, Shariati Far Fateme, Saghafi Dena, Shekarriz Amirhossein, Barati Mojdeh, Nasirimotlagh Farzaneh, Ziyadloo Fatemeh, Nekuifard Arefe, Farajee Narges, Letafat Mohammadreza, Mehri Tokmeh Marzieh, Teymoorianfard Nastaran, Massah Bita, Javidi Mohammad Amin
Department of Surgery, Shiraz University of Medical Sciences, Shiraz, Iran.
Department of Pediatrics, Tehran University of Medical Sciences, Tehran, Iran.
Galen Med J. 2024 Apr 28;13:e3347. doi: 10.31661/gmj.v13i.3347. eCollection 2024.
According to the anti-cancer impact of hypericin and naringenin, we put the main aim of this study to unravel the apoptotic/anti-cancer effect of these compounds on Y79 retinoblastoma cell line.
To calculate the 50%inhibitory concentration (IC50) of hypericin for 24 and 48 hours, XTT assay performed. Cytotoxic effect of naringenin investigated by XTT and trypan blue exclusion assay further confirmed the inhibitory impact of these agents on Y79 cells viability. Flow cytometry Annexin V/PI determined the cell death. The mRNAs expression level of Bax and Bcl-2 investigated by real-time PCR in different groups including the control, cells treated with naringenin, hypericin, or concurrent with both compounds.
The 24 and 48 hours IC50 of hypericin, calculated to be 2.5 and 1.25 (μg/ml), respectively. 50 (μg/ml) naringenin induced about 20% and 30% apoptosis in Y79 cells after 24 and 48 hours. Trypan blue staining and flow cytometry confirmed this data. Moreover, flowcytometry results, revealed that the kind cell death occurred in these cells post treatment was mostly apoptosis. Simultaneous treatment with both agents didn't show synergistic effect. Bax/Bcl-2 ratio increased in cells treated with hypericin but in cells treated with narigenin didn't show significant increase in the Bax mRNA level.
Hypericin had more cytotoxic effect in Y79 cells compared with naringenin. Furthermore, hypericin and naringenin didn't have apoptotic synergistic effect in these cells. According to the real-time PCR results, hypericin induces apoptosis in Y79 cells by disrupt the ratio of Bax/Bcl-2.
根据金丝桃素和柚皮素的抗癌作用，我们将本研究的主要目的设定为揭示这些化合物对Y79视网膜母细胞瘤细胞系的凋亡/抗癌作用。
为计算金丝桃素在24小时和48小时的50%抑制浓度（IC50），进行了XTT试验。通过XTT试验研究柚皮素的细胞毒性作用，台盼蓝排斥试验进一步证实了这些药物对Y79细胞活力的抑制作用。流式细胞术Annexin V/PI法测定细胞死亡情况。通过实时PCR检测不同组（包括对照组、用柚皮素处理的细胞、金丝桃素处理的细胞或同时用两种化合物处理的细胞）中Bax和Bcl-2的mRNA表达水平。
金丝桃素在24小时和48小时的IC50分别计算为2.5和1.25（μg/ml）。50（μg/ml）柚皮素在24小时和48小时后诱导Y79细胞凋亡约20%和30%。台盼蓝染色和流式细胞术证实了该数据。此外，流式细胞术结果显示，这些细胞处理后发生的细胞死亡类型主要是凋亡。两种药物同时处理未显示协同作用。用金丝桃素处理的细胞中Bax/Bcl-2比值升高，但用柚皮素处理的细胞中Bax mRNA水平未显示显著升高。
与柚皮素相比，金丝桃素对Y79细胞具有更强的细胞毒性作用。此外，金丝桃素和柚皮素在这些细胞中没有凋亡协同作用。根据实时PCR结果，金丝桃素通过破坏Bax/Bcl-2比值诱导Y79细胞凋亡。
