去甲斑蝥素抑制乳腺癌SKBR3细胞增殖及侵袭转移的体外研究

Effect of norcantharidin on proliferation and invasion of human breast cancer cell line SKBR3 in vitro

目的:探讨去甲斑蝥素(norcantharidin,NCTD)抑制乳腺癌细胞SKBR3增殖及侵袭转移的机制。方法:体外培养人乳腺癌SKBR3细胞株,采用MTT法检测NCTD对乳腺癌细胞SKBR3增殖抑制能力;流式细胞术测NCTD对SKBR3细胞周期和凋亡的影响;应用肿瘤细胞体外黏附实验、迁移实验和Transwell体外侵袭转移模型检测NCTD对SKBR3细胞黏附、运动、侵袭转移的抑制作用。结果:NCTD可抑制人乳腺癌SKBR3细胞增殖,具有明显量效和时间依赖性,24h的IC50为12.5mg/L;10mg/LNCTD作用SKBR3细胞24h、48h后,特异地使SKBR3细胞发生G2/M期阻滞,G2/M期细胞百分比分别为35.82%、38.70%,G1期和S期细胞较对照组明显减少,亚G1和G0峰DNA含量逐渐增加,24h、48h的凋亡率分别为3.44%和6.17%;SKBR3细胞的体外黏附、运动、侵袭转移能力随NCTD药物浓度提高而下降,表现在黏附抑制率增加,过河时间延长,过膜细胞数减少(P<0.05)。结论:NCTD可通过抑制乳腺癌细胞增殖,阻滞细胞周期,诱导细胞凋亡,抑制肿瘤细胞对基质黏附、迁移运动、侵袭转移等途径抑制癌症的发展。

AIM ： To investigate the effect of norcantharidin （NCTD） on proliferation and invasion of human breast cancer cell line SKBR3 in vitro and its anticancer mechanisms. METHODS： MTT assay was used to determine SKBR3 cell proliferation. Light and FACScan were used to detect apoptosis and cell cycle. The invasiveness of SKBR3 was evaluated by the adhesion test, Matrigel experiment and the crossing - river test. RESULTS： NCTD had inhibitive effects on growth of SKBR3 cells in a dose and time - dependent manner, with the IC50 value of 12.5 mg/L at 24 h. The cells treated with 10 mg/L NCTD for 24 h and 48 h showed typical apoptotic morphology and hypodiploid peak before G1 phase. The cell cycle was arrested at G2/M phase. The apoptosis percentage was up to 3.44% and 6. 17%, and the G2/M percentage was up to 35. 82% and 38. 70%. NCTD also could inhibit obviously the adhesion, movement and invasive capability simulating human basement membrane of SKBR3. Its effect was also in a dose - dependent manner. In the NCTD - treated group, crossing - river time was prolonged significantly and passing - membrane cells markedly decreased. CONCLUSION： NCTD in vitro inhibits not only the proliferation and growth of human breast cancer cells but also invasion and metastasis of the cells at relatively low concentration. NCTD shows prominent anti -tumor effects.