Cancer Res Treat.  2017 Jan;49(1):150-160. 10.4143/crt.2015.462.

Gastric Carcinogenesis in the miR-222/221 Transgenic Mouse Model

• Choi B
• Yu J
• Han TS
• Kim YK
• Hur K
• Kang BC
• Kim WH
• Kim DY
• Lee HJ
• Kim VN
• Yang HK

• KMID: 2367512
• DOI: http://doi.org/10.4143/crt.2015.462

Abstract

PURPOSE
MicroRNAs (miRNAs) regulate various cellular functions, including development, cell proliferation, apoptosis, and tumorigenesis. Different signatures associated with various tissue types, diagnosis, progression, prognosis, staging, and treatment response have been identified by miRNA expression profiling of human tumors. miRNAs function as oncogenes or as tumor suppressors. The relationship between gastric cancer and miRNA garnered attention due to the high incidence of gastric cancer in Asian countries. miR-222/221 expression increases in gastric tumor tissues. The oncogenic effect of miR-222/221 was previously determined in functional studies and xenograft models. In this study, transgenic mice over-expressing miR-222/221 were generated to confirm the effect of miR-222/221 on gastric carcinogenesis.
MATERIALS AND METHODS
At 6 weeks of age, 65 transgenic mice and 53 wild-type mice were given drinking water containing N-nitroso-N-methylurea (MNU) for 5 alternating weeks to induce gastric cancer. The mice were euthanized at 36 weeks of age and histologic analysis was performed.
RESULTS
Hyperplasia was observed in 3.77% of the wild-type mice and in 18.46% of the transgenic mice (p=0.020). Adenoma was observed in 20.75% of the wild-type mice and 26.15% of the transgenic mice (p=0.522). Carcinoma was observed in 32.08% of the wild-type mice and 41.54% of the transgenic mice (p=0.341). The frequency of hyperplasia, adenoma, and carcinoma was higher in transgenic mice, but the difference was statistically significant only in hyperplasia.
CONCLUSION
These results suggest that hyperplasia, a gastric pre-cancerous lesion, is associated with miR-222/221 expression but miR-222/221 expression does not affect tumorigenesis itself.

Keyword

microRNAs; Transgenic mouse; Stomach neoplasms; Gastric carcinogenesis; MNU (N-nitroso-N-methylurea)

MeSH Terms

Adenoma
Animals
Apoptosis
Asian Continental Ancestry Group
Carcinogenesis*
Cell Proliferation
Diagnosis
Drinking Water
Heterografts
Humans
Hyperplasia
Incidence
Mice
Mice, Transgenic*
MicroRNAs
Oncogenes
Prognosis
Stomach Neoplasms
Drinking Water
MicroRNAs

Figure

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  Fig. 1. Schematic diagram of the study design indicating that mice were observed for 36 weeks, followed by immunohistochemical analysis. MNU, N-nitroso–N–methylurea.

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  Fig. 2. Phenotype of miR-222/221 transgenic (TG) mice and selection. (A) miR-222/221 expression was confirmed in the stable cell line by northern blot analysis. Pre-miRNA-222/221 and mature miRNA-222/221 levels were increased in the stable cell line transfected with miR-222/221. (B) miR-222/221 expression was assessed by polymerase chain reaction analysis with the CMV promoter and miR-222 primers in offsprings to select TG or wild-type (WT) mice. Mouse 1, 2, and 4 were generated from a TG mouse. (C) miR-222/221 expression level in various organs from miR-222/221 TG and WT mice.

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  Fig. 3. Representative histological images of normal, pre-cancerous lesions, and carcinoma in transgenic and wild-type mice (A, C, D, H&E staining, ×40; B, H&E staining, ×100).

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  Fig. 4. Cancer-related markers analyzed by immunohistochemistry. (A) Histological analysis of the stomachs of MNU-treated mice. CD31 and Ki-67 expression increased in transgenic (TG) mice compared to that in wild-type (WT) mice (×100). No significant difference was observed in caspase 3, p57, and p27 expression. (B) Histological analysis of the stomachs of MNU-untreated mice. p27 expression was similar in the stomachs of WT and TG mice.

Reference

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