Experimental Study
Role of SMAD family member 4 tumor-suppressor gene in sporadic colorectal tumorigenesis of transgenic mouse model
Li Huali, Cheng Huangrong, Yang Chao, Huang Shuoyang, Zheng Yongbin
Published 2020-11-08
Cite as Chin J Exp Surg, 2020, 37(11): 2071-2074. DOI: 10.3760/cma.j.cn421213-20200624-00453
Abstract
ObjectiveTo construct ApcloxP/loxP+ Smad4loxP/loxP double transgenic mouse model which can imitate the process of tumoregenisis of human sporadic colorectal cancer (CRC) and to verify the important role of SMAD family member 4 (Smad4) gene.
MethodsThegenotypic milieuof adenomatous polyposis coli gene(Apc)tm1Tyj/J and Smad4tm2.1Cxd/Jmicewas transformed into C57BL/6J mice (all mouse were purchased from The Jackson Laboratory) and mice were then crossed to establish atransgenic mouse strain. Finally, offspring mice with ApcloxP/loxP+ Smad4loxP/loxP were generated by polymerase chain reaction (PCR). Transgenic mice and C57BL/6J mice (12, respectively) were injected with LentivirusCre-IRES-Luciferase into the intestinal mucosa under colonoscope. After intraperitoneal injection of D-luciferase, the mice were imaged through in vivo imaging system (IVIS)to observe tumor progression dynamically. And the tumor tissues were sampled and stained with Hematoxylin-Ehong (HE) to verify the tumoregenicity of mice. The t test was used to compare the two groups.
Results22 doubletransgenic mice with ApcloxP/loxP+ Smad4loxP/loxP were successfully bred, tissue neoplasia wereinduced by LentivirusCre-IRES-Luciferase and formed sporadic colorectal tumor lesions which were confirmed by IVIS and histology. At the end of 12 weekend, tumoregenesis rate was 33% (4/12), but tumor lesion was not observed in all C57BL/6J mice. The body weight of the experimental and control group were (22.58±1.21), (21.36±1.01) g, respectively, with no statistically significant difference (t=0.892, P>0.05); and the age of mice were (63±1), (62±1) d, respectively, with no statistically significant difference (t=0.121, P>0.05).
ConclusionThe transgenic mice with ApcloxP/loxP+ Smad4loxP/loxP constructed in this experiment induced tumoregenesis by LentivirusCre-IRES-Luciferase, successfully simulated the process of human sporadic CRC, and confirmed that the knockout of Smad4 gene can promote the occurrence of CRC.
Key words:
Colorectal cancer; Adenomatous polyposis coli (Apc) gene; SMAD family member 4 gene; Model, mouse
Contributor Information
Li Huali
Department of Gastointestinal Surgery, Renmin Hospital of Wuhan University, Wuhan 430064, China
Cheng Huangrong
Department of Gastointestinal Surgery, Renmin Hospital of Wuhan University, Wuhan 430064, China
Yang Chao
Department of Gastointestinal Surgery, Renmin Hospital of Wuhan University, Wuhan 430064, China
Huang Shuoyang
Department of Gastointestinal Surgery, Renmin Hospital of Wuhan University, Wuhan 430064, China
Zheng Yongbin
Department of Gastointestinal Surgery, Renmin Hospital of Wuhan University, Wuhan 430064, China
