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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

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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

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