过表达HER2增强黑素瘤B16细胞的增殖及侵袭和迁移

Over-expressed HER2 enhances proliferation, migration and invasion of melanoma B16 cells

目的构建稳定过表达人表皮生长因子受体2(HER2/ErbB2)基因的黑素瘤细胞株,研究其对黑素瘤细胞的增殖、侵袭和迁移的影响。方法将pCMV3-ErbB2(人源)重组质粒通过脂质体法转染至黑素瘤B16细胞株,用潮霉素B筛选出耐药阳性克隆。实时定量PCR(qRT-PCR)检测各单克隆ErbB2 mRNA相对表达量,免疫荧光技术检测ErbB2蛋白的表达。MTT法检测转染后细胞增殖能力,TranswellTM实验检测细胞侵袭能力,划痕实验检测细胞迁移能力。结果实时定量PCR检测结果表明,通过脂质体法成功构建过表达HER2的B16细胞株,其HER2的表达显著高于空载体对照细胞。TranswellTM等实验显示HER2过表达增强黑素瘤细胞增殖、侵袭和迁移的能力。结论过表达HER2促进黑素瘤细胞的增殖、侵袭和迁移。

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 p CMV3-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. TranswellTMassay was performed to determine the effect of HER2over-expression on the capability of B16 s 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.