羟苯基维胺脂对顺铂抗卵巢癌活性影响的研究

Effect of 4-HPR enhancing anti-cancer efficancy of cisplatin to ovarian cancer

目的:探讨羟苯基维胺脂(4-HPR)联合顺铂(DDP)对卵巢癌SKOV3细胞形态学、增殖、凋亡和细胞周期的影响,及其与血管内皮生长因子(VEGF)、尿激酶型纤溶酶原激活物(u-PA)表达变化的关系。方法:观察不同浓度4-HPR对SKOV3细胞形态学变化;MTT法检测4-HPR、DDP及联合对SKOV3细胞增殖的影响;流式细胞仪检测4-HPR、DDP对SKOV3细胞凋亡和细胞周期的影响;RT-PCR法检测4-HPR、DDP对SKOV3细胞VEGF、u-PAmRNA表达的影响。结果:1μmol/L的DDP对SKOV3细胞增殖抑制作用较弱,≥5μmol/LDDP可明显抑制SKOV3细胞增殖,P<0.05。1μmol/L的4-HPR对SKOV3细胞生长抑制作用不明显,≥2.5μmol/L时,4-HPR对SKOV3细胞生长抑制作用明显增强,P<0.05。5μmol/L的DDP联合4-HPR(1、5和10μmol/L)能显著抑制SKOV3细胞的增殖,两者联合对SKOV3细胞的抑制作用较DDP单独用药明显增强,P<0.05。不同浓度4-HPR和DDP作用SKOV3细胞48h均可诱导细胞凋亡并将细胞阻滞于G2～M期和S期,在两者联合时上述作用更加明显,P<0.05。不同浓度4-HPR和DDP作用SKOV3细胞48h均可抑制细胞VEGF、u-PAmRNA的表达,P<0.01。结论:4-HPR联合DDP能抑制卵巢癌SKOV3细胞增殖,诱导细胞凋亡,阻滞细胞周期,其机制可能与抑制VEGF、u-PA表达有关。

OBJECTIVE： To study the effect of 4-HPR combined with DDP on morphological changes, proliferation, apoptosis and cell cycle of human ovarian cancer SKOV3 cells and its relationship with VEGF and u-PA expressions. METHODS： The effect of 4-HPR on morphological changes of SKOV3 cell was observed by an optical microscope. The effect of 4-HPR and DDP on SKOV3 cell proliferation was measured by MTT assay. The cell apoptosis and cell cycle were analyzed by flow cytometry （FCM）. The VEGF and u-PA mRNA expressions of SKOV3 cells were detected by RT-PCR analysis. RESULT： One vmol/L of DDP had weak inhibitory effect on SKOV3 cells, more than 5 μmol/L of DDP had remarkable inhibitory effect on SKOV3 Cell （P〈0.05）. One μmol/L of 4-HPR had uneonspieuous inhibitory effect on SKOV3 cell, and more than 2.5 /μmol/L 4-HPR had distinct inhibitory effect on SKOV3 cell （P〈 0. 05）. Five μmol/L of DDP combined with 4-HPR （1,5 and 10μmol/L） significantly inhibited the proliferation of ovarian cancer SKOV3 cells, which had a statistical difference compared with the DDP single medication group （P〈 0.05）. 4-HPR combined with DDP induced cell apoptosis and arrested SKOV3 cell at G2-M and S stage compared with the control and 4-HPR, DDP used alone groups （P 〈 0.05）. 4-HPR combined with DDP decreased the expressions of VEGF and u-PA mRNA compared with the control and 4-HPR, DDP used alone groups （P%0.01）. CONCLUSION： 4-HPR combined with DDP can significantly inhibit the proliferation, induce apoptosis, and block cell cycle of ovarian cancer SKOV3 cells, which is associated with the VEGF and u-PA expressions.