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A characterization of four B16 murine melanoma cell sublines molecular fingerprint and proliferation behavior.

Danciu C, Falamas A, Dehelean C, Soica C, Radeke H, Barbu-Tudoran L, Bojin F, Pînzaru SC, Munteanu MF - Cancer Cell Int. (2013)

Bottom Line: 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.

View Article: PubMed Central - HTML - PubMed

Affiliation: Faculty of Pharmacy, University of Medicine and Pharmacy "Victor Babes", EftimieMurgu Square, No. 2, 300041 Timişoara, România.

ABSTRACT

Background: 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.

Methods: 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.

Results: 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.

Conclusion: Molecular fingerprint and proliferation behavior of four B16 melanoma cell sub-lines were elucidated by associating SERS investigations with MTT proliferation assay.

No MeSH data available.


Related in: MedlinePlus

Typical SERS spectra collected from the B16F10 cells upon incubation with Ag nanoparticles. Group a) and b) represents signals collected from two distinct cells, whereas c), d) and e) are spectra collected from melanoma skin tissue (B16 cells injection) incubated with nanoparticles. The spectral details of a) are shown in the right side. Excitation 633 nm. Laser power 5 mW
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Figure 5: Typical SERS spectra collected from the B16F10 cells upon incubation with Ag nanoparticles. Group a) and b) represents signals collected from two distinct cells, whereas c), d) and e) are spectra collected from melanoma skin tissue (B16 cells injection) incubated with nanoparticles. The spectral details of a) are shown in the right side. Excitation 633 nm. Laser power 5 mW

Mentions: The last analyzed cell line was B16F10. These cells presented particularly weak SERS signal. The spectra presented in Figure 5 show typical SERS signal acquired from this line. Figure 5 Typical SERS spectra collected from the B16F10 cells upon incubation with Ag nanoparticles. Group a) and b) represent signals collected from two distinct cells, whereas c-e are spectra collected from melanoma skin tissue (B16 cells injection) incubated with nanoparticles. For the investigation of this cell sub-line, the spectra were acquired by moving the laser spot from one place to another in a conglomerate of cells. The recorded signal was different compared to the previous cell lines and presented more background noise than usual. The SERS spectra collected from the two groups of cells presented here show similar vibrational modes, regardless of the fact that the spectra collected from the second group of cells, are very weak in intensity. The strongest band observed in these spectra is located at 230 cm-1. Other bands are observed at 648, 995, and the two broad bands in the fingerprint region centered at 1299, 1336, and 1525 and 1555 cm-1, respectively. Some of these bands were reported by previous SERS studies of cells, as well [10,34] and were assigned to amide II, tryptophan (1555 cm-1), CH2, CH3 bending vibrations of proteins (1336 cm-1), proteins amide III (995 cm-1), CCH bending and ring breathing vibration modes of tyrosine or O-P-O stretching vibration of DNA/RNA (830 cm-1). Based on our previous studies focusing on the investigation of the SERS signal from tissues [36], we believe that the clearly resolved bands around 1300 cm-1 are attributable to the nucleic species.


A characterization of four B16 murine melanoma cell sublines molecular fingerprint and proliferation behavior.

Danciu C, Falamas A, Dehelean C, Soica C, Radeke H, Barbu-Tudoran L, Bojin F, Pînzaru SC, Munteanu MF - Cancer Cell Int. (2013)

Typical SERS spectra collected from the B16F10 cells upon incubation with Ag nanoparticles. Group a) and b) represents signals collected from two distinct cells, whereas c), d) and e) are spectra collected from melanoma skin tissue (B16 cells injection) incubated with nanoparticles. The spectral details of a) are shown in the right side. Excitation 633 nm. Laser power 5 mW
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3750233&req=5

Figure 5: Typical SERS spectra collected from the B16F10 cells upon incubation with Ag nanoparticles. Group a) and b) represents signals collected from two distinct cells, whereas c), d) and e) are spectra collected from melanoma skin tissue (B16 cells injection) incubated with nanoparticles. The spectral details of a) are shown in the right side. Excitation 633 nm. Laser power 5 mW
Mentions: The last analyzed cell line was B16F10. These cells presented particularly weak SERS signal. The spectra presented in Figure 5 show typical SERS signal acquired from this line. Figure 5 Typical SERS spectra collected from the B16F10 cells upon incubation with Ag nanoparticles. Group a) and b) represent signals collected from two distinct cells, whereas c-e are spectra collected from melanoma skin tissue (B16 cells injection) incubated with nanoparticles. For the investigation of this cell sub-line, the spectra were acquired by moving the laser spot from one place to another in a conglomerate of cells. The recorded signal was different compared to the previous cell lines and presented more background noise than usual. The SERS spectra collected from the two groups of cells presented here show similar vibrational modes, regardless of the fact that the spectra collected from the second group of cells, are very weak in intensity. The strongest band observed in these spectra is located at 230 cm-1. Other bands are observed at 648, 995, and the two broad bands in the fingerprint region centered at 1299, 1336, and 1525 and 1555 cm-1, respectively. Some of these bands were reported by previous SERS studies of cells, as well [10,34] and were assigned to amide II, tryptophan (1555 cm-1), CH2, CH3 bending vibrations of proteins (1336 cm-1), proteins amide III (995 cm-1), CCH bending and ring breathing vibration modes of tyrosine or O-P-O stretching vibration of DNA/RNA (830 cm-1). Based on our previous studies focusing on the investigation of the SERS signal from tissues [36], we believe that the clearly resolved bands around 1300 cm-1 are attributable to the nucleic species.

Bottom Line: 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.

View Article: PubMed Central - HTML - PubMed

Affiliation: Faculty of Pharmacy, University of Medicine and Pharmacy "Victor Babes", EftimieMurgu Square, No. 2, 300041 Timişoara, România.

ABSTRACT

Background: 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.

Methods: 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.

Results: 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.

Conclusion: Molecular fingerprint and proliferation behavior of four B16 melanoma cell sub-lines were elucidated by associating SERS investigations with MTT proliferation assay.

No MeSH data available.


Related in: MedlinePlus