不同浓度松弛素-2对人永生角质细胞增殖、迁移的影响

Effects of different concentrations of Relaxin-2 on the proliferation and migration of human immortalized keratinocytes

目的观察不同浓度松弛素-2(RLN-2)对人永生角质(HaCaT)细胞增殖、迁移能力的影响。方法将HaCaT细胞在不同浓度RLN-2的培养基中培养,以不含RLN-2的培养基培养的细胞作为对照组,应用CCK-8试剂检测其对细胞增殖影响,细胞划痕实验检测其对细胞迁移能力的影响,流式细胞术检测其对细胞生长周期的影响,Western blot检测细胞周期蛋白Cyclin B1和Cyclin A2表达情况。结果以RLN-2浓度为10~100 ng/ml培养HaCaT细胞24 h后,相比对照组,增殖逐渐加快、迁移能力逐渐增强,相关细胞周期蛋白Cyclin B1和Cyclin A2表达水平增高,处于S期和G 2/M期细胞比例增加,并在100 ng/ml处达到峰值;RLN-2浓度200 ng/ml培养的HaCaT细胞相比100 ng/ml组增殖减慢、迁移能力下降,相关细胞周期蛋白Cyclin B1和Cyclin A2表达水平下降,处于S期和G 2/M期细胞比例下降。结论RLN-2在适当浓度范围内能增强HaCaT细胞的迁移能力,同时也可通过促进相关细胞周期蛋白表达,提高处于S期和G 2/M期细胞比例促进细胞增殖。

Objective To explore the effect of different concentrations of relaxin-2(RLN-2)on the proliferation and migration abilities of human immortalized keratinocytes(HaCaT cells).Methods HaCaT cells were cultured in media with different concentrations of RLN-2,and the cells were cultured in media without RLN-2 as the control group.The effect on cell proliferation was assessed by using the CCK-8 reagent,the cell migration ability was evaluated through in vitro cell scratch assay,the cell cycle was examined by flow cytometry,and the expression levels of cell cycle proteins Cyclin B1 and Cyclin A2 were detected by Western blot.Results After being cultured for 24 hours under RLN-2 concentration ranging from 10~100 ng/ml,HaCaT cells showed progressively increased proliferation and migration capabilities compared to the control group,with elevated expression levels of cell cycle proteins Cyclin B1 and Cyclin A2 and an increased proportion of cells in S and G 2/M phases,peaking at 100 ng/ml.However,HaCaT cells cultured with 200 ng/ml of RLN-2 exhibited reduced proliferation and migration capabilities,decreased expression levels of Cyclin B1 and Cyclin A2,and a lower proportion of cells in S and G 2/M phases compared to the 100 ng/ml group.Conclusion RLN-2 can enhance the migration ability of HaCaT cells within an appropriate concentration range and may also promote cell proliferation by increasing the expression of related cell cycle proteins and the proportion of cells in S and G 2/M phases.