Download Detection and Quantification of Stem Cell Biomarkers for Cancer

Investigation of Squamous Cell Cancer in Developing Zebrafish
Kiel T. Tietz and Michael A. Pickart
Zebrafish as a Human
Tumor Experimental
Model
Results
Developing embryos injected 4 hours
post-fertilization showed SCC presence
on 1 day post injection analysis, but no
SCCs were identified on day 2 - 5
analysis (As illustrated in figure 1)
SCCs within
Zebrafish
Embryo
Figure 2. a) Embryonic zebrafish b) adult zebrafish
Thirty years ago, researchers discovered
that introducing cancer cells onto
developing mice actually contributed to the
formation of a variety of normal tissues
instead of forming tumors
Recently, A study found that the zebrafish
embryonic environment may contain cues
that also suppress the tumorigenicity of
cancerous human melanoma cells
How are Tumors Grown
in Zebrafish?
Further Experimentation
Figure 1. 1 day post-injection zebrafish embryo positive for
SCC cancer cells
Hypothesis
•If squamous cell cancer is injected into the
vascular system of 2 day post-fertilization
adolescent zebrafish, then tumors will
spread and establish within the fish
•If squamous cell cancer is injected into 4
hours post-fertilization zebrafish embryos
then tumor growth will be inhibited
The Duct
of Cuvier
Figure 5. Illustration of the vascular system of a adolescent in a
fluroescent transgenic zebrafish
Conduct tumor growth analysis of 2 day postfertilization injections into the Duct of Cuvier and
study the spread of SCCs through the vasculature
of the zebrafish using fluorescent imagery
How is tumor growth analyzed?
The Big Picture
These experiments will be beneficial
to understand normal surrounding
tissue interacting with tumor cells for
head and neck & other epithelial
cancers
Figure 3. Micromanipulator on a
dissection microscope
Oral squamous cell carcinoma cancer
cells are Injected onto embryos 4
hours post-fertilization and
adolescents 2 days post-fertilization
Fluorescent cells are harvested and
suspended to 10,000,000 cells per mL of
phosphate-buffered saline solution and the
needle is calibrated to 100 SCC cancer
cells per injection
Figure 4. a) Hoechst stained SCC cancer cells b) Green fluorescent
protein stained SCC cancer cells
SCCs used for injection express green
fluorescent protein and can be easily
visualized
Images are taken with a fluorescence
microscope 1 - 5 days post injection to
analyze differences in tumor spread and
growth
Further analysis may provide an
opportunity to isolate genetic
components underlying tumor growth
and treatment response. Thus,
assisting in the development of
improved therapy
Lee, Lisa M.J, Elisabeth A. Seftor, Gregory Bonde, Robert A. Cornell,
and Mary J.C. Hendrix. "The Fate of Human Malignant Melanoma
Cells Transplanted Into Zebrafish Embryos: Assessment of Migration
and Cell Division in the Absence of Tumor Formation."
DEVELOPMENTAL DYNAMICS (2005): 1560-570. Print.