SHP2不同突变体对乳腺癌细胞迁移和侵袭能力的影响

Effect of different SHP2 mutants on migration and invasion of breast cancer cell

目的:采用慢病毒感染的方式将SHP2基因和SHP2突变体导入乳腺癌细胞系MDA-MB-231中,研究SHP2及其各突变体对乳腺癌细胞迁移和侵袭能力的影响。方法:构建SHP2-WT和SHP2-T468M、SHP2-N308D和SHP2-2YF 3个突变体的慢病毒载体,完成慢病毒包装后,感染乳腺癌细胞系MDA-MB-231稳定表达。通过蛋白免疫印记检测高表达情况,免疫荧光观察定位情况,划痕、Transwell实验检测细胞的迁移和侵袭能力。结果:SHP2-WT增强了乳腺癌细胞迁移和侵袭能力,SHP2-T468M,SHP2-N308D与对照相比无显著影响,SHP2-2YF与对照相比降低了乳腺癌细胞的迁移和侵袭能力。结论:SHP2-WT能显著增强乳腺癌细胞迁移和侵袭能力,这可能与其磷酸酶活性无关,而与其542位和580位酪氨酸的磷酸化有关。

Objective：To explore the effects of SHP2 and its mutants on migration and invasion of breast cancer cells by transfecting SHP2 and SHP2 mutants into breast cancer cell line MDA-MB-231. Methods： First inglentiviral vector SHP2-WT and three mutants of SHP2-T468 M, SHP2-N308 D and SHP2-2YF were constructed, and then the lentiviral packaging was completed to infect breast cancer cell line MDA-MB-231 for stable expression. The expression, location, cell migration and invasion were detected by western blotting,immunofluorescence, wound-healing and transwell assay respectively. Results： Migration and invasion abilities of breast cancer cell were enhanced by SHP2-WT. SHP2-T468 M, SHP2-N308 D had no significant effect compared with the control group. The SHP2-2YF lessened the migration and invasion in breast cancer cells. Conclusion： SHP2-WT can significantly increase the invasion and migration of breast cancer cells, which may not be related to the phosphatase activity, but associate with 542 and 580 tyrosine phosphorylation.