Abstract
Background and aims: Colorectal 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. Methods: In 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. Results: Our results showed the successful construction and transfection of pcDNA3 L. major Gp63 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. Conclusion: Due 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
Highlights
Colorectal cancer (CRC) is known as the third leading neoplasm in both men and women worldwide [1] which has a 12% five-year survival rate when it becomes metastatic [2]
We introduced a model to assess experimental hepatic and pulmonary metastasis of CT26 colon carcinoma cells in syngeneic BALB/c mice by systemic injection of CT26 tumor cells expressing Leishmania glycoprotein 63 (Gp63) gene into the tail vein
L. major Gp63 gene was amplified with PCR by the specific primers which had two restriction sites (HindIII and EcoRI)
Summary
Colorectal cancer (CRC) is known as the third leading neoplasm in both men and women worldwide [1] which has a 12% five-year survival rate when it becomes metastatic [2]. In Iran, CRC is the third leading cancer in men and the fourth in women [6]. Experimental models of micrometastasis are necessary for diagnostic and therapeutic investigation of tumor. The tumor cells are injected intraportally to target the liver [10]. Micrometastases are small clusters of cancer cells with no diagnostic evidence during diagnosis and surgery. We developed a mouse model to evaluate micrometastasis of colon carcinoma cells by systemic injection of tumor cells. Methods: In this study, stably transfected CT26 cells expressing Leishmania major GP63 were intravenously (IV) injected into BALB/c mice for induction of micrometastases.
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