tianjin medical university different tumorigenesis of the orthotopic and hypodermic models of...
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TIANJIN MEDICAL UNIVERSITY
Different Tumorigenesis Of The Orthotopic And Hypodermic Models
Of Papillary Thyroid Carcinoma Cells
In Nude Mice.
Ye Yan
Metabolic Diseases Hospital & Tianjin Institute of Endocrinology, Tianjin Medical
University, Tianjin, China
TIANJIN MEDICAL UNIVERSITY
BACKGROUND
Papillary thyroid carcinoma (PTC) is the most prevalent form accounting for 80% of all thyroid carcinoma.
PTC is frequently associated with RET/PTC1
rearrangement and BRAFV600E(T1799A) mutation, especially in the sporadic patients.
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OBJECTIVE
To observe and compare the different
tumorigenesis of the orthotopic and
hypodermic models of PTC cell lines
in nude mice.
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METHODS
The following human PTC cell lines were used: TPC-1, B5-16 and B2-7.
TPC-1 and B2-7---- RET/PTC1 rearrangement
B5-16 ----BRAFV600E mutation
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The Orthotopic and hypodermic nude mice models of thyroid cancer.
Experimental animal: 8-week-old female nude mice.
Anesthesia: intraperitoneal injection of chloralic hydras.
Surgery: A 1-cm-long midline incision was made, then the underlying submandibular glands and the strap muscles were bluntly dissected. The thyroid was clearly exposed.
Orthotopic and hypodermic injection: 2*105 cells in D-Hanks was injected into the left thyroid gland and under back skin of nude mice, respectively.
◆ Observation time: At 4 and 12 weeks. ◆ Observation data: The weight of orthotopic thyroid tumor
and hypodermic tumor. Serum FT3,FT4 were detected using chemiluminescent immunoassay.
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RESULTS
Marker TPC-1 B2-7 B5-16
B5-16:BRAFV600E(T1799A mutation)
GGTGATTTTGGTCTAGCTACAGAGAAATCTCGATGGAGTGGGTCCCATCAGTTTGAACAG
TPC-1:
GGTGATTTTGGTCTAGCTACAGTGAAATCTCGATGGAGTGGGTCCCATCAGTTTGAACAG
B2-7:
GGTGATTTTGGTCTAGCTACAGTGAAATCTCGATGGAGTGGGTCCCATCAGTTTGAACAG
GAPDH
RET/PTC1
Marker TPC-1 B2-7 B5-16
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The tumorigenesis of the orthotopic and hypodermic models of PTC cell lines in
nude mice was remarkable dissimilarity.
◆ TPC-1 and B5-16 were 100% tumorigenic in the orthotopic and hypodermic mice models (n=10).
◆ However, B2-7 was only 10% tumorigenic in the orthotopic model, and it was negative in the hypodermic model (n=10).
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The development of the orthotopic and hypodermic tumor in TPC-1 and B5-16 models for 4 weeks.
PTC cell line
n Weight of hypodermic tumor (mg)
Weight of orthotopic tumor (mg)
TPC-1 6 49.9±26.8 9.8±2.1
B5-16 6 3.7±0.9 58.4±19.7
TPC-1
B5-16
0
10
20
30
40
50
60
TPC-1 B5-16
hypodermictumororthotopictumor
mg
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TPC-1
B5-16
0
100
200
300
400
500
600
TPC-1 B5-16
hypodermi ctumororthotopi ctumor
mg
The development of the orthotopic and hypodermic tumor in TPC-1 and B5-16 models for 12 weeks.
PTC cell line
n Weight of hypodermic tumor (mg)
Weight of orthotopic tumor (mg)
TPC-1 10 540.2±242.7 120.3±66.4
B5-16 10 19.1±15.5 209.9±136.9
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The level of thyroid hormone reflects the destroy of thyroid function in the orthotopic models.
4 weeks 12 weeks
0
5
10
15
20
25
FT3 FT4
TPC-1B5-16Control
0
5
10
15
20
25
FT3 FT4
TPC-1B5-16Control
*
* *
*
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The morphological changes of the orthotopic thyroid tumor (HE staining)
TPC-1 B5-16
Control
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CONCLUSIONThe growth of PTC cell lines needs different
internal environment. The tumorigenesis of the orthotopic and hypodermic models
of three PTC cell lines in nude mice was different.
The hypodermic or orthotopic PTC models should be chosen according to the study objective.
The hypodermic model---Its operation and observation is easy and convenient.
The orthotopic model ---Its phenotype is close to the primary PTC.
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ACKNOWLEDGE
Professor LS.Teng and Dr WB.Wang, The First Affiliated Hospital, College of Medicine, Zhejiang University.
My teacher professor ZP.Chen ,YQ.Yan and my workmate. Metabolic Diseases Hospital & Tianjin Institute of Endocrinology, Tianjin Medical University, Tianjin, China
TIANJIN MEDICAL UNIVERSITY