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Y. Wang
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P3.02 - Biology/Pathology (ID 620)
- Event: WCLC 2017
- Type: Poster Session with Presenters Present
- Track: Biology/Pathology
- Presentations: 1
- Moderators:
- Coordinates: 10/18/2017, 09:30 - 16:00, Exhibit Hall (Hall B + C)
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P3.02-079 - A 3D Spheroid Culture Represents an Improved in Vitro Model of Malignant Plural Mesothelioma (MPM) (ID 10472)
09:30 - 09:30 | Author(s): Y. Wang
- Abstract
Background:
Most biologists rely on cell culture in the two-dimensional (2D) format for studying tumour context which does not accurately reflect the in vivo state. 3D cell culture techniques provide cell-to-cell interactions that better mimic pathological conditions such as cancer. Malignant pleural mesothelioma (MPM) is a deadly cancer with no effective treatment and is highly drug resistance. Our study has addressed this problem by growing cells in 3D thus creating an environment that is more closely mimics the realistic tumour state for molecular and cell effect studies.
Method:
MPM cell lines were grown in conventional 2D, our newly optimised 3D format, hanging drop and poly-HEMA methods. Cells were analysed their structure by light, scanning electron (SEM) and transmission electron (TEM) microscopy. Drug infiltration was confirmed by intravital-microscopy. Cell proliferationswith with different dose of drugs (Cisplatin and Gemcitabine) were analysed in 2D and 3D cells. Cells grown as truly spherical spheroids and their 2D counterparts were harvested for tumour suppressor analysis. 16 previously reported tumour suppressors (ANK1, MIB1, RGS22, TNIL, GMC, SVIL, ATG4D, HOXB4, SCLC25A13, CHST11, ATG4D, GTF2A1, KIAA1361, PDZD2, WDR1 and TMSB15B) and 3 oncogenes (YAP1, ABCG2 and YB1) were analysed.
Result:
The 3D spheroids represent an improved 'mini-tumour' model as indicated by the visualization of cell junctions under TEM. The 3D spheroids (11 MPM lines) formed using our method also provided perfect spherical shape and revealed healthy and surface morphology by SEM analysis. We showed in our model drugs was able to penetrate from outside to centre of the spheroids. However, in our hands, the hanging drop and the poly-HEMA versions did not always produce spherical 3D cells. Cells grown in our 3D model display greater drug resistance when compared with 2D cells. Most tumour suppressor biomarkers we analysed showed down-regulation of mRNA expression level compared with cells in 2D. These tumour suppressor genes were host genes of microRNAs (e.g. MIB1 for miR-1). Most of them are frequently down-regulated in MPM. Our 3D model also showed up-regulation of genes that contribute to drug resistance such as Hif1a, YAP1, ABCG2 and YB1.
Conclusion:
3D cells grown with the newly optimised method provide a better mimic of more realistic MPM evidence by the more resistance of MPM cells to cisplatin and gemcitabine when compared to cells grown in 2D. MPM cells grown in 3D also down-regulated of tumour suppressors and up-regulation of drug resistance genes when compared to 2D cells.
