Research On Cell Apoptosis Induced By Curcumin Used For High Content Screening On Microfluidics

Drug screening is important to drug discovery. High content screening(HCS) has become a research hotspot in drug discovery due to itscharacteristics of multi-target analysis and multi-parameter measurement. Inlight of its many advantages like high transparency, non-toxicity, micron scalesize, etc, microfluidic chip could provide an ideal platform for cell biologyresearch. Moreover, its operation modules can be flexibly assembled andintegrated, which makes it an suitable platform for cell-based HCS.In this paper, we first explored the potential of naturally extracteddrug-curcumin on inducing the apoptosis of human breast cancer MCF-7cellline. Further, we investigated the feasibility of using microfluidics for HCS bystudying the variation of parameters during apoptosis. The specific researchissues included:(1) The potential of curcumin on inducing the apoptosis of MCF-7through MTT and flow cytometry (FCM) was verified, then the relationshipbetween dose and effect was established with the result of FCM.(2) Three types of report plasmid: Cyt C-GFP, AIF-GFP and EndoG-GFP were adopted to transfect MCF-7cell line. The cells were thencultured in the medium containing curcumin of different concentration.Finally, the relocation of three types of report plasmid was analyzed.(3) A microfluidic chip featured by parallel gradient generation networkswas fabricated to study cell-curcumin screening, accomplished cell culture,transfection, staining and cell-curcumin interaction on this chip. Besides, thevariation of nuclear size, plasma membrane permeability and relocation ofEndo G-GFP were statistically analyzed. Multi-parameter information of cellsabout the response to the drug curcumin was obtained, which verified thefeasibility of integrating HCS on microfluidics.(4) Preliminary semi-automatic analysis on whether a single cell is apoptotic was realized with Matlab, which provided a foundation for furtherautomatic analysis of multiple cells.In summary, our experiment results confirmed that microfluidic chipcould provide a new suitable platform for HCS.