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RUNX3 Inhibits the Invasion and Metastasis of Human Colon Cancer HT-29 Cells by Upregulating MMP-2/9
Jun Xue1, Xueliang Wu1, Ming Qu1
1Department of General Surgery, First Affiliated Hospital of Hebei North University, Zhangjiakou 075000, China.
Objective:
To investigate the effect of Runt-associated transcription factor 3 (RUNX3) on the invasion and metastasis of human colon cancer HT-29 cells and to preliminarily explore the mechanism of its anticancer effect.
Methods:
The RUNX3 plasmid vector was transfected into human colon cancer HT-29 cells by liposome-mediated transfection, while the empty vector and the blank group were used as the control group. After Geneticin (G418) screening, HT-29 cells with stable expression of RUNX3 gene were obtained. The expressions of mRNA and proteins of RUNX3 and metalloproteinases (MMP)-2/9 were detected by reverse transcription-polymerase chain reaction (RT-PCR) and western blot. Cell proliferation was determined by MTT assay. The effect of RUNX3 on invasion and metastasis of HT-29 cells was evaluated by scratch injury assay, Transwell chamber, and Matrigel invasion model.
Results:
RUNX3 was expressed stably in HT-29 cells after transfection. The expressions of RUNX3 mRNA and proteins in the experimental group were significantly higher than those in the blank/empty vector groups. Meanwhile, the expressions of MMP-2/9 mRNA and proteins in the observation group were significantly lower than those in the blank group and the empty vector group. The proliferation and migration ability in the experimental group was significantly lower than blank/empty vector groups from the third day. Transwell chamber experiment and Matrigel invasion assay showed that the number of Transwell cells was decreased significantly than blank/empty vector groups, but no difference was found between the blank group and the empty vector group.
Conclusion:
RUNX3 can inhibit the invasion and metastasis of human colon cancer HT-29 cells, and the mechanism may be related to decreased expression of MMP-2 and MMP-9.
Insights
Runt-associated transcription factor 3 (RUNX3) inhibits colon cancer cell invasion and metastasis. This is linked to reduced expression of matrix metalloproteinases (MMP)-2 and MMP-9, suggesting a potential therapeutic target.
Area of Science:
- Oncology
- Molecular Biology
- Cell Biology
Background:
- Colon cancer is a leading cause of cancer-related deaths worldwide.
- Understanding the molecular mechanisms driving colon cancer invasion and metastasis is crucial for developing effective treatments.
- Runt-associated transcription factor 3 (RUNX3) is a tumor suppressor gene implicated in various cancers.
Purpose of the Study:
- To investigate the effect of RUNX3 on the invasion and metastasis of human colon cancer HT-29 cells.
- To explore the preliminary mechanism of RUNX3's anticancer effect in colon cancer.
Main Methods:
- Human colon cancer HT-29 cells were transfected with a RUNX3 plasmid vector.
- Gene expression of RUNX3 and matrix metalloproteinases (MMP)-2/9 was analyzed using RT-PCR and western blot.
- Cell proliferation, invasion, and migration were assessed using MTT assay, scratch injury assay, Transwell chamber, and Matrigel invasion models.
Main Results:
- Stable RUNX3 expression was achieved in HT-29 cells.
- RUNX3 overexpression significantly reduced the expression of MMP-2 and MMP-9 mRNA and proteins.
- RUNX3 transfection led to decreased cell proliferation, invasion, and migration capabilities.
Conclusions:
- RUNX3 effectively inhibits the invasion and metastasis of human colon cancer HT-29 cells.
- The mechanism involves the downregulation of MMP-2 and MMP-9 expression.
- RUNX3 represents a potential therapeutic target for colon cancer treatment.
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