磁性纳米顺铂微球联合磁流体热疗对卵巢癌skov-3细胞增殖、凋亡及侵袭的影响

Effects of magnetic nano-cisplatin microspheres combined with magnetic fluid hyperthermia on the proliferation,apoptosis and invasion of skov-3 cell line of ovarian cancer

目的探讨磁性纳米顺铂微球联合磁流体热疗对卵巢癌skov-3细胞增殖、凋亡及侵袭的影响,并探讨其可能的机制。方法将对数生长期的skov-3细胞分为5组,即对照组(只加入培养液)、裸药组(顺铂5μmol/L)、纳米药物组(顺铂5μmol/L+Fe3O4磁性纳米顺铂微球1 g/L)、磁热化组(Fe3O4磁性纳米顺铂微球1 g/L)、磁热组(Fe3O4磁性纳米粒子1 g/L)。对照组、裸药组、纳米药物组细胞处理后均置于培养箱培养24 h,磁热组及磁热化组置于电磁场下培养(磁流体热疗)24 h。采用MTT比色法检测细胞增殖情况,RT-PCR技术检测细胞内CD44v6、基质金属蛋白酶2(MMP-2)mRNA表达水平,Transwell小室实验检测细胞侵袭运动能力,流式细胞仪检测细胞凋亡率。结果与对照组比较,其余4组细胞抑制率、凋亡率、侵袭抑制率均升高(P均<0.05)。纳米药物组MMP-2、CD44v6mRNA表达量与对照组比较,P均>0.05;磁热组、裸药组、磁热化组中MMP-2、CD44v6mRNA表达量逐渐降低,与对照组比较,P均<0.05或<0.01(磁热化组);纳米药物组、磁热组、裸药组、磁热化组中MMP-2、CD44v6mR-NA表达量组间比较,P均<0.05。结论磁感应加热后磁性纳米顺铂微球对卵巢癌skov3细胞增殖、侵袭的抑制作用及诱导细胞凋亡的作用更强,与其降低MMP-2、CD44v6mRNA表达有关。

Objective To investigate the effects of magnetic nano-cisplatin microspheres combined with magnetic fluid hyperthermia on the proliferation,apoptosis and invasion of skov-3 cell line of ovarian cancer and the mechanism.Methods The logarithmic growth phase skov-3 cells were divided into five groups： the control group,naked drug group（cisplatin 5μmol / L）,nano drug group（cisplatin 5 μmol / L ＋ Fe3O4 magnetic nano-cisplatin microspheres 1g / L）,magnetic thermochemotherapy group（Fe3O4 magnetic nano-cisplatin microspheres 1 g / L） and magnetic heating group（Fe3O4 magnetic nanoparticles 1 g / L）.Cells in the control group,naked drug group and nano drug group were cultured in the incubator for 24 h.Cells in the magnetic thermochemotherapy group and magnetic heating group were treated by magnetic fluid hyperthermia under the electromagnetic field for 24 h.The proliferation was measured by MTT;the levels of CD44v6,MMP-2 mRNA expression were measured by RT-PCR;the invasive ability of cells was detected by transwell test;and the apoptotic rate was detected by flow cytometry（FCM）.Results Compared with the control group,inhibitory rate of cell proliferation,apoptosis rate and invasion inhibition rate in the remaining 4 groups were increased（all P 0.05）.There were no statistical significant differences in the expression of MMP-2,CD44v6 mRNA between nano drug group and control group（P 0.05）.MMP-2,CD44v6 mRNA expression of magnetic heating group,naked drug group and magnetic thermochemotherapy group was decreased gradually（P 0.05 or P 0.01）.There were statistically significant differences in MMP-2 and CD44v6 mRNA expression between the nano drug group,magnetic heating group,naked drug group and magnetic thermochemotherapy group（all P 0.05）.Conclusion Magnetic nano-cisplatin microspheres combined with magnetic fluid hyperthermia can not only enhance the inhibitory effects of proliferation and invasion,but also induce the apoptosis,which is associated with the down-regulation of MMP-2,CD 44v6 mRNA expression.