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新型光敏剂DTPP与650纳米激光联合使用可导致乳腺癌MCF-7细胞发生高效凋亡和细胞骨架塌陷。

Combination of a novel photosensitizer DTPP with 650 nm laser results in efficient apoptosis and cytoskeleton collapse in breast cancer MCF-7 cells.

作者信息

Wang H, Zhang H M, Yin H J, Zheng L Q, Wei M Q, Sha H, Li Y X

机构信息

Laboratory of Laser Medicine, Institute of Biomedical Engineering, Academy of Medical Science, Peking Union Medical College, Tianjin, 300192, China,

出版信息

Cell Biochem Biophys. 2014 Jul;69(3):549-54. doi: 10.1007/s12013-014-9830-x.

Abstract

Luminal A type breast cancer was suitable for Photodynamic therapy (PDT) as its strong adhesion ability, low malignancy and easily being exposed to laser. To examine the novel photosensitizer agent 5-5-(4-N, N-diacetoxylphenyl-10, 15, 20-tetraphenylporphyrin)(DTPP) mediate PDT in breast cancer cell, Luminal A type breast cancer MCF-7 cells were used in this study, various concentrations of DTPP (0, 2, 4, 6, 8, 10, 12, 15, 20, 25, 30 μg/mL) and different time intervals (0, 0.5, 1, 2, 4, 6, 8 min) of laser exposure at 650 nm wavelength (power of 20 mW) were tested in PDT. The survival rates of MCF-7 cells were measured using a sensitive cell proliferation assay (MTT) to establish optimal semilethal dose and optimal time exposure, a further study of effects on cytoskeleton and apoptosis were also performed. Cell cycle and apoptosis variation were assayed by flow cytometry. Microtubule, microfilament, and nuclei were observed using laser scanning confocal microscopy. Oncoproteins Bcl-2, beta-tubulin, and beta-catenin were detected by means of electrophoresis. The novel DTPP showed an efficient growth inhibition of MCF-7 during PDT, effective combinations in MCF-7 cells were shown to be 4 μg mL(-1) PS irradiated for 8 min at least or 15 μg mL(-1) irradiated for 2 min at least. Microtubule, microfilament, and nucleus staining demonstrated that cytoskeletal collapse occurs at 0.5 h after PDT. Bcl-2 and skeleton adhesion proteins beta-catenin were reduced in the level of expression; whereas, skeleton proteins beta-tubulin and actin maintained similar levels of expression 12 h after PDT. These results provided a better understanding of DTPP-PDT in MCF-7 cells.

摘要

管腔A型乳腺癌因其较强的黏附能力、低恶性程度以及易于接受激光照射,适合进行光动力疗法(PDT)。为了检测新型光敏剂5-5-(4-N,N-二乙酰氧基苯基-10,15,20-四苯基卟啉)(DTPP)介导的PDT对乳腺癌细胞的作用,本研究使用了管腔A型乳腺癌MCF-7细胞,测试了不同浓度的DTPP(0、2、4、6、8、10、12、15、20、25、30μg/mL)以及在650nm波长(功率20mW)下不同的激光照射时间间隔(0、0.5、1、2、4、6、8分钟)对PDT的影响。使用灵敏的细胞增殖检测法(MTT)测量MCF-7细胞的存活率,以确定最佳半致死剂量和最佳照射时间,还进一步研究了其对细胞骨架和细胞凋亡的影响。通过流式细胞术检测细胞周期和凋亡变化。使用激光扫描共聚焦显微镜观察微管、微丝和细胞核。通过电泳检测癌蛋白Bcl-2、β-微管蛋白和β-连环蛋白。新型DTPP在PDT过程中对MCF-7细胞显示出有效的生长抑制作用,在MCF-7细胞中的有效组合显示为至少4μg mL(-1)的光敏剂照射8分钟或至少15μg mL(-1)照射2分钟。微管、微丝和细胞核染色表明,PDT后0.5小时发生细胞骨架塌陷。Bcl-2和骨架黏附蛋白β-连环蛋白的表达水平降低;而骨架蛋白β-微管蛋白和肌动蛋白在PDT后12小时保持相似的表达水平。这些结果有助于更好地理解DTPP-PDT在MCF-7细胞中的作用。

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