醋酸甲羟孕酮通过诱导上皮-间质转化促进卵巢癌细胞转移机制研究

摘要/Abstract

摘要：

目的探讨醋酸甲羟孕酮（MPA）对人卵巢上皮性癌（EOC）细胞转移能力的影响及其作用机制。方法 2011年8月至2012年6月在河北医科大学病理学实验室进行实验。培养EOC细胞SKOV3，取对数生长期细胞用于实验，分别设立对照组和MPA组（10-5mol/L MPA作用于SKOV3细胞48 h或72 h）。采用划痕实验观察细胞迁移能力，倒置显微镜观察细胞形态，实时荧光定量PCR(Real Time-PCR)检测Snail、Slug mRNA表达水平，蛋白印迹法(Western blot)检测E-cadherin、N-cadherin和vimentin蛋白表达水平。结果划痕24h、48h时，MPA组剩余划痕面积为［（0.34 ±0.06）%、（0.14±0.02）%］均显著小于对照组［（0.51±0.05）%、（0.46±0.06）%］，差异有统计学意义（P<0.05）。与对照组相比，10-5mol/L MPA作用于SKOV3细胞72h后，细胞由上皮形态转变为间质形态，即细胞由立方形或椭圆形转变为纺锤形或多角形，且细胞间连接松散。10-5mol/L MPA作用于SKOV3细胞48h，可显著上调 Snail、Slug mRNA表达水平，与对照组相比，差异有统计学意义（P<0.05）；10-5mol/L MPA作用于SKOV3细胞72 h，E-cadherin蛋白表达水平较对照组明显减少，差异有统计学意义（P<0.05）。而N-cadherin和vimentin蛋白表达水平较对照组明显增加，差异均有统计学意义（P<0.05）。 结论 MPA可通过诱导EOC细胞发生上皮-间质转化，进而增强EOC细胞的转移能力。

关键词: 卵巢肿瘤, 醋酸甲羟孕酮, 上皮-间质转化

Abstract:

Abstract： Objective To explore the effect of medroxyprogesterone acetate (MPA) on human epithelial ovarian cancer (EOC) cell metastasis. Methods The experiment was done at the pathology laboratory in Hebei Medical University during the August 2011 to June 2012. EOC cell line SKOV3 was cultured, and assigned to control group, MPA group（cells treated with 10-5 mol/L MPA for 48h or 72h）. The effect of MPA on cell metastasis was observed by the scratch assay, morphological change was observed by phase contrast microscope, the expression of Snail and Slug mRNA was detected by Real-Time Quantitative Polymerase Chain Reaction (Real Time PCR), the expression of E-cadherin, N-cadherin and vimentin protein was detected by Western blot. Results After scratching for 24h and 48h, the remaining wound area rates of MPA group ［(0.34 ±0.06)%、(0.14±0.02）%］were significantly less than control group［（0.51±0.05)%、(0.46±0.06）%］(P<0.05). Compared with control group, treated with 10-5 mol/L MPA for 72h, the morphology of SKOV3 cell changed from epithelial cuboidal or cobbly morphology to spindle-shaped or polygonal morphology, and increased intercellular separation. Compared with control group, the expression of Snail and Slug mRNA in SKOV3 cell were significantly up-regulated after treated with 10-5 mol/L MPA for 48h (P<0.05). Treatment of 10-5 mol/L MPA for 72h could down-regulate the E-cadherin protein, meanwhile up-regulate the N-cadherin and vimentin protein. Compared with control group, there was significant difference, respectively (P<0.05). Conclusion MPA prompted the metastasis of EOC cell through inducing the epithelial-mesenchymal transition.

Key words: ovarian neoplasms, medroxyprogesterone acetate, epithelial-mesenchymal transition

中图分类号:

R737.3

闫晓楠，徐春琳,宋阳阳，王惠兰. 醋酸甲羟孕酮通过诱导上皮-间质转化促进卵巢癌细胞转移机制研究[J]. 中国实用妇科与产科杂志, 2013, 9(10): 789-793.

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