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首页> 外文期刊>Photodiagnosis and Photodynamic Therapy >Immunocytochemical studies on the nuclear ubiquitous casein and cyclin-dependent kinases substrate following 5-aminolevulinicacid-mediated photodynamic therapy on MCF-7 cells
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Immunocytochemical studies on the nuclear ubiquitous casein and cyclin-dependent kinases substrate following 5-aminolevulinicacid-mediated photodynamic therapy on MCF-7 cells

机译：5-氨基乙酰丙酸介导的光动力疗法对MCF-7细胞核无处不在的酪蛋白和细胞周期蛋白依赖性激酶底物的免疫细胞化学研究

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Background: Recent data indicates that nuclear ubiquitous casein and cyclin-dependent kinases substrate (NUCKS) may play role in tumor growth. In present study authors examined whether photodynamic therapy with 5-aminolevulinic acid (5-ALA) induces NUCKS expression in breast cancer cell line, MCF-7. Methods: In the experiment concentration of 5-ALA was 6.5 mM. Excitation wavelength was 630±20nm, total light dose of light 5 or 10J/cm~2 and irradiance 60mW/cm~2 was used. Cells were collected at established time points and Western blot and immunocytochemical studies were performed using antibody against NUCKS. Results: Studies proved strong cytotoxic effects in cells following PDT with 6.5 mM of precursor and 10J/cm~2. Western blot analysis revealed the strongest expression of NUCKS at 7h after PDT. At next time points, 18 and 24 h, expression of NUCKS decreased and became similar to that of control group. Further immunocytochemical studies showed very strong expression of NUCKS following PDT with 5-ALA and light irradiation of 5 J/cm~2. Early, at 0h, that expression was predominantly seen in nuclei, while at 7h expression of NUCKS was observed in disseminated manner within entire cells in both nuclei and cytoplasm, with prevalence of cytoplasmic staining. Conclusions: Authors suggest that NUCKS is involved in cellular responses following PDT, and since parallel induction of NUCKS and proapoptotic marker Bax and inhibition of anti-apoptotic Bcl-2 was observed, this protein might also be involved in induction of apoptosis following PDT.
机译：背景：最近的数据表明，核无处不在的酪蛋白和细胞周期蛋白依赖性激酶底物（NUCKS）可能在肿瘤生长中起作用。在本研究中，作者研究了用5-氨基乙酰丙酸（5-ALA）进行光动力疗法是否能在乳腺癌细胞MCF-7中诱导NUCKS表达。方法：在实验中5-ALA的浓度为6.5 mM。激发波长为630±20nm，总光剂量为5或10J / cm〜2，辐照度为60mW / cm〜2。在确定的时间点收集细胞，并使用抗NUCKS抗体进行蛋白质印迹和免疫细胞化学研究。结果：研究证明，PDT对6.5 mM前体和10J / cm〜2的细胞具有强烈的细胞毒性作用。 Western blot分析显示PDT后7h NUCKS表达最强。在下一个时间点18和24小时，NUCKS的表达降低并且变得类似于对照组。进一步的免疫细胞化学研究显示，PDT联合5-ALA和5 J / cm〜2的光照射后，NUCKS的表达非常强。早期，在0h，该表达主要在细胞核中观察到，而在7h，在核和细胞质的整个细胞中以扩散的方式观察到NUCKS的表达，普遍存在细胞质染色。结论：作者建议NUCKS参与PDT后的细胞反应，并且由于观察到NUCKS和促凋亡标记Bax的平行诱导和抗凋亡Bcl-2的抑制，该蛋白也可能参与PDT诱导的细胞凋亡。

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