AI Article Synopsis
- The study aimed to create a stable B16 melanoma cell line that over-expresses the HER2 gene to investigate its impact on cell behavior.
- The researchers used techniques like plasmid transfection, PCR, and various assays to confirm HER2 over-expression and measure changes in proliferation, migration, and invasion rates of the cells.
- Results showed that over-expressing HER2 significantly increased the growth, migration, and invasion abilities of B16 melanoma cells.
Article Abstract
Objective To establish a melanoma B16 cell line stably over-expressing the HER2 (ErbB2) gene and study its effect on the proliferation, migration and invasion of melanoma cells. Methods The recombinant plasmid pCMV3-ErbB2 was transfected into B16 melanoma cell line by cationic liposome. The positive clones were screened with hygromycin B. Real-time quantitative PCR (qRT-PCR) was utilized to determine the expression of ErbB2 mRNA. Immunofluorescence was used to evaluate the expression of ErbB2 protein. MTT assay was applied to detect the cell proliferation ability. The cell migration ability was examined by a scratch assay. Transwell assay was performed to determine the effect of HER2 over-expression on the capability of B16s invasion. Results The cell line over-expressing HER2 was successfully established as showed by qRT-PCR. Over-expression of HER2 remarkably enhanced the proliferation, invasion and migration of B16 cells. Conclusion Over-expression of HER2 can enhance the growth and invasion of B16 cells.
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