SP600125对人宫颈癌HeLa细胞增殖和侵袭的影响

目的 探讨SP600125对人宫颈癌HeLa细胞的增殖周期、凋亡以及侵袭的影响.方法 采用CCK-8法检测不同时间点不同浓度的SP600125作用后HeLa细胞的增殖状态.确定20μmol/L的SP600125用于后续实验.利用平板克隆形成实验检测细胞增殖能力,DAPI染色观察细胞核形态,流式细胞仪检测细胞周期和凋亡,...

Effects of SP600125 on Proliferation and Invasion of Human Cervical Cancer HeLa Cells

Objective To investigate the effect of SP600125 on the proliferation, cell cycle, apoptosis and invasion of human cervical cancer HeLa cells. Methods CCK-8 method was used to detect the proliferation of HeLa cells treated with different concentrations of SP600125 at different time points. The 20 μmol/L of SP600125 was determined for subsequent experiments. Cell proliferation ability was detected using plate clone formation assay; nuclear morphology was observed by DAPI staining; cell cycle and apoptosis were measured by flow cytometry; cell migration and invasion were detected by cell scratch and Transwell methods; the mRNA and protein levels of p53, Mad2L1 and CDC20 were measured by qRT-PCR and Western blot after SP600125 treatment at different time points. Results Compared with control group (0.1%DMSO), cells proliferative activity were reduced by 10, 20, 30, 40 and 50 μmol/L SP600125 treatment for 24h. Compared with control group, the rate of apoptosis was significantly increased in SP600125 treatment groups, and the cell proportion in G₂/M phase increased (P<0.001), while the cell proportion in G₀/G₁ phases cells was reduced after SP600125 treatment for 24h and 48h (P<0.001), and the clonal number, migration and invasion ability of HeLa cells also decreased significantly (P<0.001). qRT-PCR and Western blot results showed a significant decrease in Mad2L1 mRNA and protein expression (P<0.05) and a significant increase in p53 and CDC20 mRNA and protein expression (P<0.01). Conclusion SP600125 can induce cell cycle arrest of cervical cancer HeLa cells in G2/M phase by upregulating p53 and CDC20 and downregulating Mad2L1 expression, and promote cell apoptosis to inhibit cell proliferation, migration and invasion.