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目的观察不同化疗耐药性卵巢癌细胞的侵袭转移特性,并探讨其可能机制。方法体外建立卵巢癌不同梯度顺铂(DDP)耐药细胞系模型(OVCAR-3/DDP-1、OVCAR-3/DDP-2、OVCAR-3/DDP-3);MTT法、流式细胞仪、Transwell小室观察亲本细胞和耐药细胞的增殖、细胞周期分布、细胞侵袭和迁移等生物学特性的变化;ELISA法测定细胞培养上清液中基质金属蛋白酶-2(MMP-2)、组织金属蛋白酶抑制因子-2(TIMP-2)、基质金属蛋白酶-9(MMP-9)、组织金属蛋白酶抑制因子-1(TIMP-1)的浓度。结果成功建立不同梯度顺铂耐药细胞系模型,OVCAR-3/DDP-1、DDP-2、DDP-3细胞的耐药指数依次为3.87、8.39、13.42。耐药细胞均较亲本细胞增殖缓慢,G0/G1期细胞比例增加,S期细胞比例减少,其中OVCAR-3/DDP-2、DDP-3与亲本细胞比较差异有统计学意义(P<0.05)。与亲本细胞相比,耐药细胞伴随其耐药性的增加,侵袭转移能力逐渐增强,其中OVCAR-3/DDP-2、DDP-3与亲本细胞相比差异有统计学意义(P<0.05)。细胞上清液中MMP-2/TIMP-2、MMP-9/TIMP-1的比值随顺铂耐药性的增加而逐渐上升,耐药细胞与亲本细胞相比差异均有统计学意义(P<0.05)。结论对顺铂耐药性的产生和增加可促进卵巢癌细胞的侵袭和转移,可能部分与耐药细胞MMP-2/TIMP-2、MMP-9/TIMP-1比值上调有关。
Objective To observe the invasion and metastasis of different chemotherapy resistant ovarian cancer cells and to explore its possible mechanism. Methods DDP-resistant cell lines (OVCAR-3 / DDP-1, OVCAR-3 / DDP-2 and OVCAR-3 / DDP-3) were established in vitro. MTT assay, flow cytometry Transwell chamber was used to observe the changes of biological characteristics such as cell proliferation, cell cycle distribution, invasion and migration of parental cells and drug-resistant cells. The levels of matrix metalloproteinase-2 (MMP-2) (TIMP-2), matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) Results The model of cisplatin-resistant cell lines was established successfully. The resistance index of OVCAR-3 / DDP-1, DDP-2 and DDP-3 cells were 3.87, 8.39 and 13.42. Compared with the parental cells, the proliferation of drug-resistant cells was slower, the proportion of cells in G0 / G1 phase increased, and the proportion of cells in S phase decreased. There was significant difference between OVCAR-3 / DDP- . Compared with the parental cells, the resistant cells increased gradually with the increase of drug resistance, the ability of invasion and metastasis increased gradually. The difference between OVCAR-3 / DDP-2 and DDP-3 was statistically significant (P <0.05) . The ratio of MMP-2 / TIMP-2 and MMP-9 / TIMP-1 in the cell supernatant gradually increased with the increase of cisplatin resistance, and there was significant difference between the drug-resistant cells and the parental cells <0.05). Conclusions The generation and increase of resistance to cisplatin can promote the invasion and metastasis of ovarian cancer cells, which may be partly related to the up-regulation of the ratio of MMP-2 / TIMP-2 and MMP-9 / TIMP-1.