Molecular Beacon Imaging to Visualize Ki67 mRNA for Cell Proliferation Ability

Area of Science:

• Biotechnology
• Cell Biology
• Nanomedicine

Background:

• Cell proliferation is a fundamental biological process crucial for development and tissue repair.
• Accurate visualization of cell proliferation in living cells remains a challenge.
• Molecular beacons (MB) offer potential for detecting specific mRNA targets, but efficient cellular delivery is needed.

Purpose of the Study:

• To develop and validate a novel system for visualizing cell proliferation in real-time using molecular beacons (MB).
• To investigate the efficacy of cationized gelatin nanospheres (cGNS) for enhancing MB internalization and tracking proliferation markers.
• To assess the correlation between basic fibroblast growth factor (bFGF) concentration and Ki67 mRNA expression as a measure of cell proliferation.

Main Methods:

• Two types of MBs were synthesized: one targeting Ki67 mRNA (proliferation marker) and another targeting GAPDH mRNA (control).
• MBs were encapsulated within cationized gelatin nanospheres (cGNS) to improve cellular uptake.
• KUM6 cells were treated with varying concentrations of bFGF after incubation with cGNS-MBs, and fluorescence was monitored using time-lapse imaging.

Main Results:

• cGNS effectively delivered both Ki67 MB and GAPDH MB into KUM6 cells without significant differences in physicochemical properties or uptake.
• Fluorescence intensity of cGNS-Ki67 MB significantly increased with increasing bFGF concentrations, indicating enhanced cell proliferation.
• Fluorescence of cGNS-GAPDH MB remained constant, confirming the specificity of the Ki67 MB response to bFGF.
• Time-lapse imaging demonstrated a rapid increase in cGNS-Ki67 MB fluorescence upon bFGF addition.

Conclusions:

• The cGNS-MB system provides a reliable method for the chronological visualization of cell proliferation ability in living cells.
• This approach enables real-time monitoring of cellular responses to growth factors like bFGF.
• The developed system holds promise for applications in regenerative medicine and drug discovery research.