A 3D Co-culture Spheroid Screening System for Studying the Malignant Transformation of Ductal Carcinoma in situ to Invasive Ductal Carcinoma Restricted; Files Only
Jiao, Qiao (Spring 2023)
Published
Abstract
Ductal carcinoma in situ (DCIS) is a precursor of invasive ductal carcinoma (IDC). Although only a small number of patients develop the invasive disease, most DCIS cases undergo aggressive treatments. Therefore, it is urgent to find alternative treatments that can effectively intercept the malignant transformation of DCIS into IDC to alleviate the therapeutic burden and avoid overtreatment. Previous studies demonstrated that a subgroup of DCIS cells is genetically and phenotypically altered, and these alterations may play a crucial role in IDC transition. Thus, a deeper understanding of the mechanistic drivers of DCIS malignant transformation into IDC is required for developing effective therapeutic interventions. In this study, an in vitro fluorescence-labeled 3D co-culture spheroid system that mimics the DCIS-to-IDC transition was utilized to identify anti-cancer compounds that could prevent the malignant transition. In this 3D spheroid co-culture assay, the proliferation rate and the scale of tumor microinvasion of MCF10CA1a (DCIS)-mCherry cells were significantly reduced after separate treatments with paclitaxel, Olaparib, doxorubicin, and cisplatin, while MCF10A-GFP cells (non-tumorigenic ductal epithelium) remained unaffected. Overall, the 3D DCIS co-culture spheroid assay proved to be a viable and physiologically relevant assay platform to mimic the ductal structure and perform high throughput drug screenings. Future studies will continue to focus on elucidating mechanistic and molecular drivers of DCIS progression into IDC.
Table of Contents
INTRODUCTION 1
DCIS Incidence and Prevalence 2
DCIS Treatments and Clinical Challenges 3
DCIS Etiology and Malignant Transformation 5
Spheroid as a Model System 8
Scope of the Thesis 10
MATERIALS AND METHODS 14
Cell Culture 15
Generating Fluorescence-labeled Stable Cell Lines with Lentivirus 15
3D Spheroid Co-culture Model 16
Anti-cancer Drug Screening and Analysis 17
RESULTS 19
Establishment of Fluorescence-Labeled Cell Lines 20
Establishment of Co-culture Spheroids 21
Phenotypic Characterization of Co-culture Spheroids Under Anti-Cancer Drug Treatments 24
DISCUSSION 44
Future Directions 41
REFERENCES 43
APPENDIX 52
About this Master's Thesis
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