双氢青蒿素对上皮性卵巢癌细胞黏附、迁移及侵袭的影响

Tan Xian-Jie, Lang Jing-He, Plouet Jean, Wu Ming, Shen Keng

Department of Obstetrics and Gynecology, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing 100730, China.

Zhonghua Yi Xue Za Zhi. 2008 Oct 14;88(37):2642-6.

OBJECTIVE

To investigate the effects of dihydroartiminisin (DHA) on the adhesion, migration, and invasion ovarian cancer cells.

METHODS

Human ovarian cancer cells of the lines SKOV3 and OVCAR3 were cultured. Suspensions of SKOV3 and OVCAR3 cells were treated with DHA of the concentrations of 0.5, 2.5, 12.5, and 62.5 micromol/L respectively, and then inoculated on the plate coated with Matrigel. MTT method was used to -determine the adhesion rate. Transwell membrane chamber model was used to evaluate the effect of DHA on the migration and invasion of the SKOV3 and OVCAR3 cells. Western blotting and reverse transcriptase polymerase chain reaction were used to detect the effect of DHA on the phosphorylation of focal adhesion kinase (FAK) and on the effect of expression of metal matrix proteinases (MMPs) and their tissue inhibitors (TIMPs) respectively.

RESULTS

(1) Compared to the cells without DHA treatment, the cell adhesion ability levels of the SKOV3 and OVCAR3 cells treated with 12.5 micromol/L DHA decreased by 76.1% and 57.9% respectively (P < 0.05), while their migration ability levels decreased by 59.3% and 69.7% respectively (P < 0.05). (2) Both SKOV3 and OVCAR3 showed weak invasion ability, and DHA only showed a slight inhibitory effect on the cell invasion of these 2 lines (both P > 0.05). (3) Compared to the cells without DHA treatment, the phosphorylation level of FAK of the SKOV3 and OVCAR3 cells treated with 12.5 micromol/L DHA decreased by 42.9% and 44.8% respectively (both P < 0.05). (4) RT-PCR showed mRNA expression of MMP2, TIMP1, and TIMP2, but not mRNA expression of MMP9 in both SKOV3 and OVCAR3 cells. The mRNA expression levels of the SKOV3 and OVCAR3 cells treated with 12.5 micromol/L DHA increased by 1.5 and 2.6 times respectively (both P < 0.05).

CONCLUSION

DHA has inhibitory effects on the adhesion and migration of epithelial ovarian cancer cells, which may be related to its down-regulation of the phosphorylation of FAK in these cells.

目的

研究双氢青蒿素（DHA）对卵巢癌细胞黏附、迁移及侵袭的影响。

方法

培养人卵巢癌细胞系SKOV3和OVCAR3。将SKOV3和OVCAR3细胞悬液分别用浓度为0.5、2.5、12.5和62.5 μmol/L的DHA处理，然后接种于包被基质胶的平板上。采用MTT法测定黏附率。利用Transwell膜小室模型评估DHA对SKOV3和OVCAR3细胞迁移及侵袭的影响。分别采用蛋白质免疫印迹法和逆转录聚合酶链反应检测DHA对黏着斑激酶（FAK）磷酸化的影响以及对金属基质蛋白酶（MMPs）及其组织抑制剂（TIMPs）表达的影响。

结果

（1）与未用DHA处理的细胞相比，用12.5 μmol/L DHA处理的SKOV3和OVCAR3细胞的细胞黏附能力水平分别降低了76.1%和57.9%（P < 0.05），而它们的迁移能力水平分别降低了59.3%和69.7%（P < 0.05）。（2）SKOV3和OVCAR3均表现出较弱的侵袭能力，DHA对这两种细胞系的细胞侵袭仅表现出轻微的抑制作用（均P > 0.05）。（3）与未用DHA处理的细胞相比，用12.5 μmol/L DHA处理的SKOV3和OVCAR3细胞的FAK磷酸化水平分别降低了42.9%和44.8%（均P < 0.05）。（4）RT-PCR显示SKOV3和OVCAR3细胞中均有MMP2、TIMP1和TIMP2的mRNA表达，但无MMP9的mRNA表达。用12.5 μmol/L DHA处理的SKOV3和OVCAR3细胞的mRNA表达水平分别增加了1.5倍和2.6倍（均P < 0.05）。