氧化偶氮甲烷-硫酸葡聚糖硫酸钠诱导小鼠结直肠肿瘤方法的优化

doi:10.24920/003495

Abstract

Since azoxymethane (AOM)-dextran sodium sulfate (DSS) induced tumorigenesis was used to explore inflammation-associated carcinogenesis of sporadic colorectal cancer (CRC), different administration modes of AOM or DSS have been reported. In this article we optimized the protocol of the AOM-DSS modeling using C57BL/6 mice for study on sporadic CRC by intraperitoneal injecting AOM solution at a proper concentration with a 100 μl sterile syringe once, feeding with DSS solution for 7 days in a roll and change DSS solution every day. More than 100 C57BL/6 mice had been treated with the optimized protocol, and all mice were demonstrated suffering from colorectal tumors when sacrificed in 8 to 20 weeks after AOM injection. These tumors mainly occurred in distal segment of colorectum with an increase in tumor density, which was similar to CRC in human beings. Tumor per mouse was high, and variation of tumor number per mouse was low. The histology of tumor developed through the defined stage ranged from precursor lesions, adenomatous lesions, adenomas to adenocarcinomas. The modified protocol of AOM-DSS model is easy, cheap, with high tumor formation rate of colorectal tumors.

Key words: animal model, colorectal cancer, inflammation, protocol

Funding: Supported by the Hebei Provincial Research Foundation of Health and Family Planning Commission (No. 20180688).

Liang Xi, Hu Jingnan, He Jianming. An Optimized Protocol of Azoxymethane-Dextran Sodium Sulfate Induced Colorectal Tumor Model in Mice[J].Chinese Medical Sciences Journal, 2019, 34(4): 281-288.

Figures/Tables 6

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Table 1

Table 2

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References 27.

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Time after AOM injection	Number of mice	Number of mice with tumors	Tumor number per mouse^§
8 W	6	6	5.17±0.95
10 W	6	6	6.83±0.79
20 W	6	6	11.83±1.58

Problem	Possible reasons	Solutions	Step
1. High death rate of mice during AOM and DSS treatment
	1) Mouse strain highly susceptible to AOM	-Switch to a strain of lower susceptibility	1
		-Decrease AOM dose	6
	2) Injury of blood vessels and/or abdominal viscera	-Ensure precise injection technique	7
	3) AOM injected into vein	-Ensure precise injection technique	7
2. High death rate of mice after DSS treatment
	1) Mouse strain highly susceptible to DSS	-Switch to a mouse strain of lower susceptibility	1
		-Decrease DSS concentration and/or reduce the time period of administration	10-14
	2) Too many and/or too large colorectal tumorous lesions	-Decrease AOM dose	6
		-Decrease DSS concentration and/or reduce the time period of administration	10-14
		-Reduce the waiting time period	15
3. Low tumor incidence and/or no or few tumors per mouse
	1) Mouse strain lowly susceptible to AOM/DSS	-Switch to a mouse strain of highly susceptibility	1
		-Increase AOM dose	6
		-Increase DSS concentration and/or prolong the period of administration	10-15
	2) AOM dose too low	-Increase AOM dose	10
	3) DSS concentration too low	-Increase DSS concentration	10-14
	4) Waiting period too short	-Prolong waiting period	15
4.High variability within experimental groups
	1) Genetic backgrounds not pure	-Purify genetic background	1
	2) Differences in origin	-Use mice from the same offspring	1
	3) Differences in gender	-Do not use female mice	1
	4) Affect by menstrual period	-Do not use female mice	1
	5) Imprecise injection	-Ensure precise injection technique	7

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