Related Experiment Video
Updated: May 19, 2026
Experimental Metastasis Assay
Published on: August 25, 2010
[Cloning and functional analysis of the tumor metastasis related gene tropomyosin2]
Yi Zhang1, Wen-Tao Liu, Qiang Li
1Department of Surgery, Ruijin Hospital, Shanghai Jiao Tong University, Shanghai Institute of Digestive Surgery, Shanghai 200025, China.
Objective:
To construct an eukaryotic expression vector of tumor associated gene tropomyosin 2 (TM2) tagged with green fluorescence protein and study its biological role in tumorigenesis. METHODS; Whole length of TM2 cDNA sequence was amplified from human gastric cancer cell line AGS by reverse transcript PCR. The recombinant TM2 was inserted into eukaryotic expression vector pIRES2 tagged with enhanced green fluorescence protein. pIRES2-EGFP was transfected into human hepatoma cell line QGY-7703 cells by liposome technique and detected by fluorescence microscopy and flow cytometry. TM2 protein level was determined by Western blot. The in vitro invasion and motility were further studied. Its tumorigenesis was assessed on nude mice.
Results:
The recombinant TM2 was shown to be expressed in QGY-7703 cells by fluorescence microscopy and flow cytometry. The protein level of TM2 was increased. The invasion ability of TM2- transfected cells was increased(45.6 +/- 9.9) than that in controls(21.6 +/- 3.3, P < 0.05), as well as mobility(41.4 +/- 11.8 vs. 16.7 +/- 3.7). The tumor volume was (241.5 +/- 95.1) mm3, significantly higher than that in blank-vector controls (100.6 +/- 85.4) mm3 and negative-controls (123.7 +/- 92.3) mm3.
Conclusion:
TM2 plays a role in growth and metastasis of hepatic tumor.
Insights
Tropomyosin 2 (TM2) promotes the growth and metastasis of hepatic tumors. This study constructed a TM2 expression vector to investigate its role in tumorigenesis, revealing increased invasion and tumor volume in transfected cells.
Area of Science:
- Molecular biology
- Cancer research
- Gene expression
Context:
- Tumorigenesis involves complex genetic alterations.
- Tropomyosin 2 (TM2) is implicated in cancer.
- Understanding TM2's role is crucial for hepatic tumor research.
Purpose:
- To construct an eukaryotic expression vector for tumor-associated gene Tropomyosin 2 (TM2) fused with enhanced green fluorescence protein (EGFP).
- To investigate the biological role of TM2 in the tumorigenesis of human hepatoma cells (QGY-7703).
Summary:
- Tropomyosin 2 (TM2) cDNA was amplified and inserted into the pIRES2-EGFP vector.
- The recombinant vector was transfected into QGY-7703 cells, confirming TM2 expression via fluorescence microscopy and flow cytometry.
- Western blot analysis showed increased TM2 protein levels.
- In vitro studies demonstrated enhanced cell invasion and motility in TM2-transfected cells.
- Tumorigenesis assays in nude mice revealed significantly larger tumor volumes in TM2-expressing groups compared to controls.
Impact:
- TM2 significantly enhances the invasion and motility of hepatic cancer cells.
- Increased TM2 expression correlates with accelerated tumor growth in vivo.
- These findings highlight TM2 as a potential therapeutic target for hepatic tumors.
More Related Videos
07:23A Protocol for Genetic Induction and Visualization of Benign and Invasive Tumors in Cephalic Complexes of Drosophila melanogaster
Published on: September 11, 2013
09:37Defining Gene Functions in Tumorigenesis by Ex vivo Ablation of Floxed Alleles in Malignant Peripheral Nerve Sheath Tumor Cells
Published on: August 25, 2021
Related Concept Videos
02:07Tumor Progression
Colon cancer is one of the best-documented examples of tumor progression. Early mutation in the APC gene in colon cells causes a small growth on the colon wall called a polyp. With time, this polyp grows into a benign, pre-cancerous tumor. Further...
02:30Metastasis
Epithelial-to-Mesenchymal Transition
The epithelial-to-mesenchymal transition or EMT is a developmental process commonly observed in wound healing, embryogenesis, and cancer metastasis. EMT is induced by transforming growth factor-beta (TGF-β) or receptor tyrosine kinase (RTK) ligands, which further...
01:33Cancer-Critical Genes I: Proto-oncogenes
When the function of certain critical genes, especially those involved in cell cycle regulation and cell growth signaling cascades, gets disrupted, it upsets the cell cycle progression. Such cells with unchecked cell cycles start proliferating uncontrollably and eventually develop into tumors.
Such genes that act...
03:03mTOR Signaling and Cancer Progression
The mTOR pathway or the...
01:05Cancer-Critical Genes II: Tumor Suppressor Genes
When the function of certain critical genes, especially those involved in cell cycle regulation and cell growth signaling cascades, gets disrupted, it upsets the cell cycle progression. Such cells with unchecked cell cycles start proliferating uncontrollably and eventually develop into tumors.
Such genes that act...