Sign In

A subscription to JoVE is required to view this content. Sign in or start your free trial.

In This Article

  • Summary
  • Abstract
  • Introduction
  • Protocol
  • Representative Results
  • Discussion
  • Acknowledgements
  • Materials
  • References
  • Reprints and Permissions

Summary

This protocol provides a mouse model of ulcerative coloproctitis-associated colorectal cancer induced by azomethane combined with dextran sulfate sodium. The model was used to evaluate the efficacy of traditional Chinese medicine compounds in the prevention and treatment of colorectal cancer.

Abstract

Colorectal cancer (CRC) is a common malignancy of the digestive system and has become the third most common malignancy worldwide and the second leading cause of malignancy-related death. Ulcerative coloproctitis (UC) is a precancerous lesion, and UC-associated CRC (UC-CRC) is the most common subtype of CRC. Therefore, a reasonable UC-CRC model is the cornerstone and guarantee of new drug development. Traditional Chinese medicine (TCM) has been widely used in the treatment of UC-CRC due to its good efficacy. As a classic tonic prescription of TCM, Liujunzi decoction (LJZD) has been widely used in the treatment of UC-CRC. In this study, a UC-CRC model was established by combining azomethane and dextran sulfate sodium, and the LJZD was administered. The data confirmed that LJZD can effectively inhibit cancer transition in UC-CRC by using mouse body weight, colorectal length, pathological and inflammatory factors, colorectal barrier function, and cancer markers. This protocol provides a system for evaluating the efficacy of TCM in the prevention and treatment of UC-CRC.

Introduction

Colorectal cancer (CRC) is a common gastrointestinal malignancy, the third most common malignancy, and the second most common cause of death in the world, accounting for 10% of the global cancer incidence and 9.4% of the total cancer-related death1,2. Genetic factors, chronic inflammation, high-fat diet, diabetes, and abnormal intestinal flora are risk factors for CRC3,4. Among them, inflammatory bowel disease, especially ulcerative coloproctitis (UC), is a clear risk factor for CRC5,6. UC-asso....

Protocol

The animal procedure has been approved by the Ethics Committee of Changchun University of Chinese Medicine (Record number: 2021214). Specific pathogen-free C57BL/6J mice (8-10 weeks, weight 18-22 g), male and female, were housed in independently ventilated cages at 22 °C and 65% relative humidity. The mice began the experiment after 7 d of adaptive feeding, during which they had free access to water and diet.

1. Drug preparation

  1. Preparation of LJZD
    R.......

Representative Results

The decoction of LJZD was prepared according to the composition ratio of drugs in Table 1 and the decoction method of TCM in Figure 1A. According to the time point indicated in Figure 1B, mice were intraperitoneally injected with 1 mg/mL AOM on the 7th day, and mice were given free access to drinking water containing 2% DSS in the 3rd, 6th, and 9th weeks. The UC-CRC mouse model was successfully establi.......

Discussion

CRC is one of the most common cancers worldwide, with approximately 1,148,000 new cases and more than 576,000 deaths each year. CRC can be divided into three types according to different causes, including hereditary, sporadic and UC-CRC31. The incidence of CRC in patients with inflammatory bowel diseases such as UC is significantly higher than that in the general population. UC stimulates the development of CRC through the inflammatory-cancer pathway, which differs from the typical adenoma-adenoca.......

Acknowledgements

This work was supported by the Jilin Provincial Department of Science and Technology (YDZJ202201ZYTS181).

....

Materials

NameCompanyCatalog NumberComments
AzoxymethaneSigmaA5486
5-amino salicylic acidKuihua Pharmaceuticals Group Jiamusi Luling Pharmaceutical Co., Ltd3819413
C57BL/6J miceLiaoning Changsheng Biotechnology Co., LtdNO 210726210100853716
Cover slipJiangsu Shitai Experimental Equipment Co., Ltd10212432C
DAB color development kitJiangsu Shitai Experimental Equipment Co., Ltd2005289
Dewatering machine Wuhan Junjie Electronics Co., LtdJJ-12J
Dextran sulfate sodiumDalian Meilun Biotechnology Co., LtdMB5535
Embedding machineWuhan Junjie Electronics Co., LtdJB-P5
Hematoxylin-eosin dyeWuhan Hundred Degree Biotechnology Co., LtdB1000
IL-6Jiangsu Meimian Industrial Co., LtdMM-0163M2
IsofluraneRWD Life Science Co., LtdR510-22-10
KI67 primary antibodyGoogle Biotechnology IncGB121141
Neutral gumWuhan Hundred Degree Biotechnology Co., Ltd10004160
Object slideJiangsu Shitai Experimental Equipment Co., Ltd10212432A
Occludin primary antibodyAffnityDF7504
Orthostatic optical microscopeNikonNikon Eclipse CI
Pathological microtomeShanghai Leica Instrument Co., LtdRM2016
ZO-1 primary antibodyAbcamab221547

References

  1. Sung, H., et al. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 Cancers in 185 Countries. CA Cancer J Clin. 71 (3), 209-249 (2021).
  2. Tsai, K. Y., et al.

Explore More Articles

Coloproctitis CancerMouse ModelTraditional Chinese MedicineLiujunzi DecoctionUlcerative ColoproctitisColorectal CancerPrecancerous LesionAzomethaneDextran Sulfate SodiumColorectal Barrier FunctionCancer Markers

This article has been published

Video Coming Soon

JoVE Logo

Privacy

Terms of Use

Policies

Research

Education

ABOUT JoVE

Copyright © 2024 MyJoVE Corporation. All rights reserved