Radiobiological effects of wound fluid on breast cancer cell lines and human-derived tumor spheroids in 2D and microfluidic culture

Abstract

Intraoperative radiotherapy (IORT) could abrogate cancer recurrences, but the underlying mechanisms are unclear. To clarify the effects of IORT-induced wound fluid on tumor progression, we treated breast cancer cell lines and human-derived tumor spheroids in 2D and microfluidic cell culture systems, respectively. The viability, migration, and invasion of the cells under treatment of IORT-induced wound fluid (WF-RT) and the cells under surgery-induced wound fluid (WF) were compared. Our findings showed that cell viability was increased in spheroids under both WF treatments, whereas viability of the cell lines depended on the type of cells and incubation times. Both WFs significantly increased sub-G1 and arrested the cells in G0/G1 phases associated with increased P16 and P21 expression levels. The expression level of Caspase 3 in both cell culture systems and for both WF-treated groups was significantly increased. Furthermore, our results revealed that although the migration was increased in both systems of WF-treated cells compared to cell culture media-treated cells, E-cadherin expression was significantly increased only in the WF-RT group. In conclusion, WF-RT could not effectively inhibit tumor progression in an ex vivo tumor-on-chip model. Moreover, our data suggest that a microfluidic system could be a suitable 3D system to mimic in vivo tumor conditions than 2D cell culture.

Biological and clinical review of IORT-induced wound fluid in breast cancer patients

“…IORT-seroma-treated and non-IORT-seroma-treated cells showed no significant change in expression levels of proteins associated with cell cycle arrest (P16, P21) and the expression level of Caspase 3. Furthermore, our results confirmed the previous findings about tumor progressive effects of seroma on these three BC cell lines ( 124 ). Figure 5 presents the studies related to the effects of seroma on BC cell lines.…”

“…It is worth noting that in both IORT-seroma and non-IORT-seroma groups, the expression levels of both E-cadherin and Caspase 3 were significantly higher in seroma-treated spheroids compared to RPMI-treated spheroids (as control). This study suggested IORT-seroma as a fluid containing inhibitory factors for tumor migration in the microfluidic system ( 124 ). Also, we showed increased proliferative and migrative characteristics of spheroids from four BC patients under IORT-seroma treatment using time-lapse imaging ( 127 ).…”

“…Spheroids derived from each patient were treated for six days with the 24h seroma collected from the patients. Final data from fluorescent live/dead staining on day 6 showed that in 22 samples, the percentage of live cells was significantly higher in seroma-treated samples compared to cells treated with Roswell Park Memorial Institute (RPMI) (as a control for each sample) ( 124 ).…”

Microfluidic Synthesis of Ultrasmall Chitosan/Graphene Quantum Dots Particles for Intranasal Delivery in Alzheimer's Disease Treatment

“…The staining process proceeded with incubation of cells with the dyes for 45 min at 37 ◦ C in the tissue culture incubator and then visualized under a fluorescence microscope. [ 48b,95 ]…”