A new in vivo model for analysis of colon carcinoma micrometastasis in BALB/c mice

Roya Abedizadeh, H. Rezvan, A. Nourian
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Abstract

Background and aimsColorectal cancer (CRC) is known as the fourth leading cause of death across the world. The fate of patients depends on the metastatic spread of cancer cells. Micrometastases are small clusters of cancer cells with no diagnostic evidence during diagnosis and surgery. Therefore, experimental models for micrometastasis are necessary to investigate tumors. We developed a mouse model to evaluate micrometastasis of colon carcinoma cells by systemic injection of tumor cells.MethodsIn this study, stably transfected CT26 cells expressing Leishmania major GP63 were intravenously (IV) injected into BALB/c mice for induction of micrometastases. The mice were divided into three groups and the groups were sacrificed on days 3, 7, and 14 of the injection. reverse transcriptase-polymerase chain reaction (RT-PCR) was performed on tissue samples to detect Gp63 gene.ResultsOur results showed the successful construction and transfection of pcDNA3 L. majorGp63 into CT26 cells. After IV injection, total cellular RNA was extracted and the Gp63 gene was detected in the liver, lung, and kidney but not in the colon.ConclusionDue to the significance of micrometastasis and the need to establish simple models for cancer research, an experimental mouse model was developed. CT26 tumor cells stably expressing Gp63 generated a potent system for diagnosis of micrometastatic cells in tissues. Injection into the tail vein is a practical model for cancer research because of the lower fatality rate and no need for anesthesia.
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一种用于分析BALB/c小鼠结肠癌微转移的新体内模型
背景和目的结直肠癌(CRC)被认为是全球第四大死亡原因。病人的命运取决于癌细胞的转移扩散。微转移是指在诊断和手术中没有诊断证据的小簇癌细胞。因此,建立微转移的实验模型是研究肿瘤的必要条件。我们建立了小鼠模型,通过全身注射肿瘤细胞来评估结肠癌细胞的微转移。方法将稳定转染的表达利什曼原虫GP63的CT26细胞静脉注射到BALB/c小鼠体内,诱导其微转移。将小鼠分为三组,分别于注射后第3、7、14天处死。采用逆转录聚合酶链反应(RT-PCR)检测Gp63基因。结果pcdna3l . majorGp63成功构建并转染至CT26细胞。静脉注射后提取细胞总RNA,在肝、肺和肾中检测到Gp63基因,但在结肠中未检测到。结论考虑到微转移的重要意义和建立肿瘤研究简单模型的需要,我们建立了小鼠实验模型。稳定表达Gp63的CT26肿瘤细胞产生了一个有效的组织微转移细胞诊断系统。尾静脉注射是一种实用的癌症研究模型,因为它的致死率较低,而且不需要麻醉。
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