过表达HER2增强黑素瘤B16细胞的增殖及侵袭和迁移

发布时间：2018-10-10 14:39

【摘要】：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。
[Abstract]:Objective to construct a melanoma cell line stably expressing human epidermal growth factor receptor 2 (HER2/ErbB2) gene and study its effect on the proliferation, invasion and migration of melanoma cells. Methods pCMV3-ErbB2 recombinant plasmid was transfected into melanoma B16 cell line by liposome method, and hygromycin B was used to screen the drug resistant clones. The relative expression of each monoclonal ErbB2 mRNA was detected by real-time quantitative PCR (qRT-PCR), the expression of ErbB2 protein was detected by immunofluorescence, the proliferative ability of transfected cells was detected by MTT assay, the ability of cell invasion was detected by TranswellTM assay, and the ability of cell migration was detected by scratch assay. Results the results of real-time quantitative PCR analysis showed that the expression of HER2 in B16 cell line which overexpressed HER2 was significantly higher than that in blank vector control cells by liposome method. TranswellTM and other experiments showed that HER2 overexpression enhanced the proliferation of melanoma cells. The ability to invade and migrate. Conclusion overexpression of HER2 promotes the proliferation, invasion and migration of melanoma cells.
【作者单位】：新乡医学院生命科学技术学院河南省高校合成生物改造工程与应用重点实验室;
【基金】：河南省重大科技专项研究课题(161100310900) 新乡医学院研究生科研创新支持计划(YJSCX201634Y)
【分类号】：R739.5

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