Faculty of Pharmacy, University of Medicine and Pharmacy "Victor Babes", EftimieMurgu Square, No. 2, 300041 Timişoara, România.
Biomedical Physics, Biomedical, Theoretical Physics, and Molecular Spectroscopy Department, Faculty of Physics, Babes-Bolyai University, Kogalniceanu 1, RO 400084 Cluj-Napoca, România.
Cancer Cell Int. 2013 Jul 26;13:75. doi: 10.1186/1475-2867-13-75. eCollection 2013.
One of the most popular and versatile model of murine melanoma is by inoculating B16 cells in the syngeneic C57BL6J mouse strain. A characterization of different B16 modified cell sub-lines will be of real practical interest. For this aim, modern analytical tools like surface enhanced Raman spectroscopy/scattering (SERS) and MTT were employed to characterize both chemical composition and proliferation behavior of the selected cells.
High quality SERS signal was recorded from each of the four types of B16 cell sub-lines: B164A5, B16GMCSF, B16FLT3, B16F10, in order to observe the differences between a parent cell line (B164A5) and other derived B16 cell sub-lines. Cells were incubated with silver nanoparticles of 50-100 nm diameter and the nanoparticles uptake inside the cells cytoplasm was proved by transmission electron microscopy (TEM) investigations. In order to characterize proliferation, growth curves of the four B16 cell lines, using different cell numbers and FCS concentration were obtained employing the MTT proliferation assay. For correlations doubling time were calculated.
SERS bands allowed the identification inside the cells of the main bio-molecular components such as: proteins, nucleic acids, and lipids. An "on and off" SERS effect was constantly present, which may be explained in terms of the employed laser power, as well as the possible different orientations of the adsorbed species in the cells in respect to the Ag nanoparticles. MTT results showed that among the four tested cell sub-lines B16 F10 is the most proliferative and B164A5 has the lower growth capacity. Regarding B16FLT3 cells and B16GMCSF cells, they present proliferation ability in between with slight slower potency for B16GMCSF cells.
Molecular fingerprint and proliferation behavior of four B16 melanoma cell sub-lines were elucidated by associating SERS investigations with MTT proliferation assay.
在同基因 C57BL6J 小鼠中接种 B16 细胞是最受欢迎和用途最广泛的鼠黑色素瘤模型之一。对不同 B16 修饰的细胞亚系进行特征描述将具有真正的实际意义。为此，采用表面增强拉曼光谱/散射（SERS）和 MTT 等现代分析工具来表征所选细胞的化学成分和增殖行为。
从四种 B16 细胞亚系（B164A5、B16GMCSF、B16FLT3、B16F10）中记录了高质量的 SERS 信号，以便观察亲本细胞系（B164A5）与其他衍生的 B16 细胞亚系之间的差异。将细胞与直径为 50-100nm 的银纳米粒子孵育，并通过透射电子显微镜（TEM）研究证明了纳米粒子在细胞质内的摄取。为了表征增殖，使用不同的细胞数量和 FCS 浓度获得了四种 B16 细胞系的生长曲线，通过 MTT 增殖测定法。为了进行相关性分析，计算了倍增时间。
SERS 带允许在细胞内识别主要的生物分子成分，如蛋白质、核酸和脂质。始终存在“开”和“关”的 SERS 效应，这可以根据所使用的激光功率以及吸附物质在细胞中的可能不同取向来解释。MTT 结果表明，在四种测试的细胞亚系中，B16 F10 增殖能力最强，B164A5 生长能力最低。关于 B16FLT3 细胞和 B16GMCSF 细胞，它们具有介于两者之间的增殖能力，B16GMCSF 细胞的增殖能力稍慢。
通过将 SERS 研究与 MTT 增殖测定法结合，阐明了四种 B16 黑色素瘤细胞亚系的分子指纹和增殖行为。
