共培养体系中小鼠囊胚对人卵巢癌细胞系HO-8910PM的影响

Effect of Mouse Blastocyst on Invasion and Metastasis of Human Ovarian Carcinoma Cells in Co-Culture System

目的:观察两种具有侵袭特性的小鼠囊胚细胞与人卵巢癌细胞在相同的体外微环境下,动态的相互作用和整体生物学行为变化。方法:建立小鼠囊胚与人高转移卵巢癌细胞系HO-8910PM体外共培养模型,应用MTT法检测HO-8910PM细胞黏附率的改变;应用Transwell体外细胞侵袭实验观察HO-8910PM细胞体外侵袭及趋化性运动能力的变化;应用RT-PCR检测HO-8910PM细胞基质金属蛋白酶(MMP)-2、MMP-9、金属蛋白酶组织抑制剂(TIMP)-1、TIMP-2mRNA的表达。结果:癌细胞与小鼠囊胚共培养24h后,大部分囊胚开始边脱带边黏附,推开底层的癌细胞,在囊胚与肿瘤细胞间形成了明显的界限。共培养组的HO-8910PM细胞的黏附率低于对照组。Transwell小室体外侵袭实验及体外趋化性运动实验显示,共培养组穿过滤膜的细胞数明显少于对照组。共培养组MMP-2、MMP-9mRNA的表达及MMP/TIMP较对照组明显下降,TIMP-1、TIMP-2mRNA的表达略有增加。结论:当人HO-8910PM细胞与小鼠囊胚共培养时,肿瘤细胞失去了特有的侵袭性;小鼠囊胚能显著降低HO-8910PM细胞的黏附、体外侵袭能力、运动迁移性及MMP-2、MMP-9mRNA的表达。

Objective：The establish the model of mouse blastocyst co-cultured with ovarian carcinoma cells and explore dynamic interaction and difference between the two kinds of life cells under the same microenvironment in vitro thereof. Methods：The model of mouse blastocyst co-cultured with highly metastatic ovarian carcinoma cell HO-8910PM was established to observe the interaction and the whole biological behavior changes of the two kinds of life cells under the same microenvironment in vitro. MTT assay was used to examine the changes of HO-8910PM cell adhesion; Transwell chamber assay was performed to determine its effect on invasion and migration capacities of HO-8910PM; and mRNA levels of MMP-2,MMP-9,TIMP-1 and TIMP-2 were assessed by RT-PCR. Results：After 24 h of being co-cultured,mouse blastocyst escaped from zone pellucida and hatched on the underlying malignant ovarian tumor cells,and then gradually pushed underlying highly proliferative and divisive tumor cells and then freely invaded them. However,the ovarian tumor cells only accumulated in the periphery of trophoblast cells as rotundity or ellipse shape. MTT assay revealed that compared with control group,the adhesive rate was significantly decreased in co-cultured group. After co-culture with mouse blastocysts,the number of HO-8910PM cells to penetrate polycarbonates coated with or without matrigel was significantly lower. After co-culture with mouse blastocysts,HO-8910PM mRNA levels of MMP-2 and MMP-9 were significantly lower; on the contrary,mRNA levels of TIMP-1 and TIMP-2 were increased; ratios of MMP/TIMP were significantly lower than those of control. Conclusion：When the model of mouse blastocysts,co-cultured with highly metastatic ovarian carcinoma cell HO-8910PM,was established,the characteristic invasion was lost in tumor cells. After being co-cultured,invasiveness of HO-8910PM cells,invasion-related adhesive and mobile capacity in vitro were decreased by mouse blastocyst. The expression of MMP-2 and MMP-9 mRNA of HO-8910PM was also down-regulated by blastocyst.