Abstract
Traditional in vitro anticancer drug sensitivity testing at the population level suffers from lengthy procedures and high false positive rates. To overcome these defects, we built a confocal Raman microscopy sensing system and proposed a single-cell approach via Raman-deuterium isotope probing (Raman-DIP) as a rapid and reliable in vitro drug efficacy evaluation method. Raman-DIP detected the incorporation of deuterium into the cell, which correlated with the metabolic activity of the cell. The human non-small cell lung cancer cell line HCC827 and human breast cancer cell line MCF-7 were tested against eight different anticancer drugs. The metabolic activity of cancer cells could be detected as early as 12 h, independent of cell growth. Incubation of cells in 30% heavy water (D2O) did not show any negative effect on cell viability. Compared with traditional methods, Raman-DIP could accurately determine the drug effect, meanwhile, it could reduce the testing period from 72–144 h to 48 h. Moreover, the heterogeneity of cells responding to anticancer drugs was observed at the single-cell level. This proof-of-concept study demonstrated the potential of Raman-DIP to be a reliable tool for cancer drug discovery and drug susceptibility testing.
Highlights
The discovery, screening, and administration of safe and effective anticancer drugs are vital for tackling cancers
It is extensively carried out during drug development to screen candidates entering clinical trials and before medical treatment to find the right medication for individual patients when many drugs become ineffective due to drug resistance
To establish the method for deuterium labeling and SCRS acquisition of cancer cell lines, HCC827 and MCF-7 cells were incubated in media containing D2 O, and their SCRS
Summary
The discovery, screening, and administration of safe and effective anticancer drugs are vital for tackling cancers. In vitro drug testing is a key step in evaluating the efficacy of anticancer drugs. It is extensively carried out during drug development to screen candidates entering clinical trials and before medical treatment to find the right medication for individual patients when many drugs become ineffective due to drug resistance. In vitro drug screening is traditionally performed on cell lines by examining their viability or proliferation after exposure to drugs. These methods detect the cell viability at the population level by measuring cellular oxidoreductase (e.g., NADP(H) and dehydrogenase) activity or adenosine triphosphate (ATP) synthesized only in viable cells. Common methods of in vitro screening include a colorimetric tetrazolium reduction (MTT) assay or other commercially available derivatives such as cell counting kit-8 (CCK-8) or CellTiter-Glo kit (CTG) [3]
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