Laser Research Centre, Faculty of Health Sciences, University of Johannesburg, Doornfontein 2028, South Africa.
Biomed Res Int. 2017;2017:5407012. doi: 10.1155/2017/5407012. Epub 2017 Sep 25.
The sensitivity of human melanoma cells to photoactivated Hypericin (Hyp) compared to aluminium(III) phthalocyanine chloride tetrasulphonate (AlPcSCl) is reported in this study. Melanoma cells (A375 cell line) were treated with various concentrations of Hyp or AlPcSCl alone, for 1, 4, and 24 hrs; varying doses of laser irradiation alone (594 or 682 nm); or optimal concentrations of PSs combined with laser irradiation. Changes in cell morphology, viability, membrane integrity, and proliferation after treatment of cells were determined using inverted microscopy, Trypan blue cell exclusion, Lactate Dehydrogenase (LDH) membrane integrity, and adenosine triphosphate (ATP) cell proliferation assay, respectively. More than 60% of cell survival was observed when cells were treated with 2.5 M of Hyp or AlPcSCl alone at all incubation times or with 5 J/cm of 594 or 682 nm laser alone. Combination of PSs and respective lasers leads to a statistically significant incubation time-dependent decrease in survival of cells. Flow cytometry using the FITC Annexin V/PI apoptosis kit demonstrated that cell death induced after Hyp-PDT is via early and late apoptosis whereas early apoptosis was the main mechanism observed with AlPcSCl-PDT. Hyp-PDT compared to AlPcSCl-PDT is indicated to be a more effective cancer cell death inducer in melanoma cells.
本研究报告了人类黑色素瘤细胞对光活化金丝桃素(Hyp)与铝(III)酞菁四磺酸钠(AlPcSCl)的敏感性比较。将黑色素瘤细胞(A375 细胞系)用不同浓度的 Hyp 或 AlPcSCl 单独处理 1、4 和 24 小时;单独用不同剂量的激光照射(594 或 682nm);或用最佳浓度的 PS 与激光照射联合处理。用倒置显微镜、台盼蓝细胞排斥试验、乳酸脱氢酶(LDH)膜完整性和三磷酸腺苷(ATP)细胞增殖测定法分别测定细胞处理后细胞形态、活力、膜完整性和增殖的变化。当细胞用 2.5M 的 Hyp 或 AlPcSCl 单独处理时,在所有孵育时间或用 5J/cm 的 594 或 682nm 激光单独处理时,观察到超过 60%的细胞存活。PSs 与相应激光联合使用会导致细胞存活率随孵育时间的推移呈统计学意义上的显著下降。用 FITC 膜联蛋白 V/PI 凋亡试剂盒进行的流式细胞术表明,Hyp-PDT 诱导的细胞死亡是通过早期和晚期凋亡,而 AlPcSCl-PDT 观察到的主要机制是早期凋亡。与 AlPcSCl-PDT 相比,Hyp-PDT 被指示为黑色素瘤细胞中更有效的癌细胞死亡诱导剂。