Genetics and Cytology Department, Genetic Engineering and Biotechnology Division, National Research Center, Dokki 12622, Cairo, Egypt.
Photodiagnosis Photodyn Ther. 2013 May;10(2):194-202. doi: 10.1016/j.pdpdt.2012.11.006. Epub 2012 Dec 5.
Photodynamic therapy (PDT) is used for the treatment of many types of predominantly epithelial cancers. Photosensitizer is taken up by fast growing tumor cells more actively than by other body cells and is activated by light, generating reactive oxygen species that cause cell death by necrosis or apoptosis. This study aimed to evaluate the efficacy of PDT with indocyanine green (ICG) through the investigation of TP53, HER-2 and TOP2A genes signals as breast cancer gene markers by interphase fluorescence in situ hybridization (nuc-FISH).
The photosynthetizer ICG (200 μM) was applied to breast cancer cell line MCF-7 cells (adenocarcinoma) in combination with laser irradiation (807 nm) exposure for 20 min and then incubated for 12, 24 and 48 h. Cell viability was evaluated using trypan blue. The signals for nuc-FISH was investigated and counted for probes specific for the genes TP53 (17p13), HER-2 (17q11.2-q12), and TOP2A (17q21-q22), and BAC-probes RP11-746M1 in 17p11.2 and RP11-403E9 in 17q11.2.
The cell viability of MCF-7 did not reduced significantly when the cells were treated with ICG (200 μM) or exposed to laser irradiation for 20 min followed by incubation for 24 h. ICG/PDT treatment with laser irradiation exposure for 20 min reduced the cell viability after incubating cells for 12, 24 and 48 h highly significantly in a time dependent manner. For nuc-FISH analysis, TP53, HER-2, TOP2A, RP11-746M1 and RP11-403E9 signals did not reduce or increase in a significant manner when the cells were treated with ICG or exposed to laser irradiation for 20 min then incubated for 24h. PDT enhanced amplification of TP53 signals from nuc ish 17p13(TP53×2) to nuc ish 17p13(TP53×3) or nuc ish 17p13(TP53×4). However, the signals of HER-2 gene, TOP2A gene and BAC probes were reduced highly significantly when MCF-7 cells were treated with PDT with all time intervals.
ICG/PDT and laser induced cytotoxic effect in MCF-7 cells. Also, PDT enhanced TP53 gene amplification, and reduced HER-2, TOP2A, and BAC probes RP11-746M1 and RP11-403E9 signals. Therefore ICG/PDT can be used for breast cancer treatment. It has the potential to induce apoptotic effect and reduce HER-2 and TOP2A genes propagation. Further in vivo studies are needed to evaluate ICG/PDT as a promising therapeutic approach for breast cancer.
光动力疗法（PDT）用于治疗多种主要上皮癌。敏化剂比其他体细胞更积极地被快速生长的肿瘤细胞摄取，并在光照下被激活，产生活性氧物种，通过坏死或凋亡导致细胞死亡。本研究旨在通过间期荧光原位杂交（nuc-FISH）评估作为乳腺癌基因标记物的 TP53、HER-2 和 TOP2A 基因信号的吲哚菁绿（ICG）PDT 的疗效。
将光合染料 ICG（200μM）应用于乳腺癌细胞系 MCF-7 细胞（腺癌），并结合激光照射（807nm）暴露 20 分钟，然后孵育 12、24 和 48 小时。使用台盼蓝评估细胞活力。研究并计数针对基因 TP53（17p13）、HER-2（17q11.2-q12）和 TOP2A（17q21-q22）以及 BAC 探针 RP11-746M1 在 17p11.2 和 RP11-403E9 在 17q11.2 的 nuc-FISH 探针信号。
当细胞用 ICG（200μM）处理或用激光照射 20 分钟后孵育 24 小时时，MCF-7 细胞的细胞活力没有显著降低。用激光照射 20 分钟进行 ICG/PDT 处理后，细胞孵育 12、24 和 48 小时后，细胞活力显著降低，呈时间依赖性。对于 nuc-FISH 分析，当细胞用 ICG 处理或用激光照射 20 分钟后孵育 24 小时时，TP53、HER-2、TOP2A、RP11-746M1 和 RP11-403E9 信号没有显著减少或增加。PDT 增强了 nuc ish 17p13（TP53×2）到 nuc ish 17p13（TP53×3）或 nuc ish 17p13（TP53×4）中 TP53 信号的扩增。然而，当 MCF-7 细胞用 PDT 处理时，HER-2 基因、TOP2A 基因和 BAC 探针的信号高度显著降低。
ICG/PDT 和激光诱导 MCF-7 细胞的细胞毒性作用。此外，PDT 增强了 TP53 基因扩增，并降低了 HER-2、TOP2A 和 BAC 探针 RP11-746M1 和 RP11-403E9 的信号。因此，ICG/PDT 可用于乳腺癌治疗。它具有诱导凋亡作用和降低 HER-2 和 TOP2A 基因传播的潜力。需要进一步的体内研究来评估 ICG/PDT 作为治疗乳腺癌的一种有前途的方法。
