Hepatitis B virus induced coupling of deadhesion and migration of HepG2 cells on thermo-responsive polymer.
The unique physical property of thermo-responsive polymer (TRP) has recently prompted its increasing applications in tissue engineering. On the other hand, TRP has not been exploited for potential applications in quantitative cell screening against external stimulations. In this study, TRP is applied as a model system for elucidating the effect of HBV replication on the biophysical responses of HepG2 cells transfected by wild type HBV genome. Moreover, mutant HBV genome is designed to assess the specific activity of the SH3-binding domain of HBx during HBV replication. The adhesion contact recession and geometry transformation of HepG2 cells transfected with empty vector (pcDNA3.1 cells), wild type HBV (wtHBV cells) and mutant HBV genome (mHBV cells) are probed during the thermal transformation across lower solution critical temperature of TRP. In comparison with pcDNA3.1 cells and mHBV cells, the initial rate of reduction in degree of deformation and average adhesion energy for wtHBV cells is significantly increased. Interestingly, migration speed and persistence time of cells are found to be correlated with the cell deadhesion kinetics. Immuno-fluorescence microscopy demonstrates that HBV replication reduces the actin concentration and focal adhesions at cell periphery during the initial 30 min cell deadhesion. The results strongly suggested that HBV infection triggers the dynamic responses of HepG2 cells through the cytoskeleton remodeling and subsequent mechanochemical transduction. Overall, it is shown that TRP provides a convenient platform for quantifying biological stimulations on adherent cells.
Li
Xi
X
Center of Biotechnology, School of Chemical and Biomedical Engineering, Nanyang Technological University, Singapore 637459, Singapore.
Feng
Huixing
H
Chen
Wei Ning
WN
Chan
Vincent
V
D000199:Actins / Q000378:metabolism; D002448:Cell Adhesion / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects*; D004247:DNA / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D005455:Fluorescent Antibody Technique; D016679:Genome, Viral / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000378:metabolism; D056945:Hep G2 Cells; D006515:Hepatitis B virus / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008856:Microscopy, Fluorescence; D010957:Plasmids / Q000235:genetics; D011108:Polymers / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013696:Temperature; D014162:Transfection
D000199:Actins; D011108:Polymers; D049452:Green Fluorescent Proteins; D004247:DNA