J4 ›› 2012, Vol. 38 ›› Issue (4): 709-712.

• 基础研究 • 上一篇    下一篇

LPA1介导溶血磷脂酸对卵巢癌细胞系基质金属蛋白酶2和9释放与活化的调控作用

汪萍1|吴小华1|单保恩2   

  1. 1.河北医科大学第四医院妇产科|河北 石家庄050011;2.河北医科大学第四医院科研中心|河北 石家庄050011
  • 收稿日期:2011-07-27 出版日期:2012-07-28 发布日期:2012-07-28
  • 通讯作者: 吴小华(Tel: 0311-87998114,E-mail:xiaohuawu65@yahoo.com) E-mail:xiaohuawu65@yahoo.com
  • 作者简介:汪 萍(1968-)|女|河北省秦皇岛市人|副主任医师| 医学博士|主要从事妇科肿瘤的临床及基础研究。
  • 基金资助:

    河北省科技厅科技攻关计划项目资助课题(JB2008-3)

Regulation of |LPA1 -mediated lysophosphatidic acid on |activities and protein expressions of MMP-2 and MMP-9
in ovarian cancer cells

WANG Ping1,WU Xiao-hua1,SHAN Bao-en2   

  1. 1.Department of Obstetrics and Gyneocology,Fourth Affiliated Hospital,Hebei Medical University,Shijiazhuang 050011,China;2.Center of Scientific Research,Fourth Affiliated Hospital,Hebei Medical University,Shijiazhuang 050011,China
  • Received:2011-07-27 Online:2012-07-28 Published:2012-07-28

摘要:

目的: 将溶血磷脂酸(LPA)受体LPA1基因的质粒载体导入卵巢癌细胞系SKOV3和3AO,建立高表达LPA1的卵巢癌细胞株,观察转染细胞株的生物学特性,探讨LPA对稳定表达LPA1的人卵巢癌细胞株基质金属蛋白酶2和9(MMP-2和MMP-9)表达及活性的影响。方法: 用脂质体介导法将pcDNA3.1flag- LPA1转入人卵巢癌细胞系(SKOV3和3AO),同时设细胞对照组及空载体组。利用RT-PCR鉴定转染细胞株表达水平,并通过检测细胞株生长、增殖变化观察转染细胞株的生物学特性。明胶酶谱检测LPA对稳定表达LPA1的卵巢癌细胞株。MMP-2和MMP-9表达及活性的影响。结果:LPA1基因的质粒载体成功导入卵巢癌细胞系SKOV3和3AO,建立高表达LPA1的卵巢癌细胞株,转染前后细胞生物学特性无明显变化。利用RT-PCR半定量检测转染细胞LPA1 mRNA表达水平,经转染后的SKOV3-LPA1 和 3AO-LPA1细胞LPA1 mRNA表达水平明显高于对照组SKOV3、3AO细胞及空载体组SKOV3-Vector、3AO-Vector细胞LPA1 mRNA的表达水平,差异有统计学意义 (P<0.05),对照组3AO、SKOV3细胞与空载体组SKOV3-Vector、3AO-Vector细胞的表达水平比较差异无统计学意义 (P>0.5)。明胶酶谱法检测结果表明,实验组SKOV3-LPA1和3AO-LPA1细胞MMP-9、MMP-2释放和活化量与对照组SKOV3、3AO和空载体组SKOV3-Vector比较,差异无统计学意义(P>0.05)。结论:经鉴定获得了生物学特性稳定的LPA1高表达的卵巢癌细胞株。在LPA诱导的MMPs的活化过程中,LPA1可能作为一个负性调节因子。

关键词: 溶血磷脂酸受体;基质金属蛋白酶;卵巢肿瘤

Abstract:

Abstract:Objective To establish an ovarian cancer cells with high expression of LPA1 by transfecting LPA1 genes into the ovarian carcer cell lines and to study thebiological characteristics of the transfected cells and to study the effects of lysophosphatidic acid (LPA) on the expressions and activities of MMP-2 and MMP-9 inovarian cancer cells with stable expression LPA1 genes.Methods The pcDNA3.1-LPA1 was transfected into ovarian cancer cells line (SKOV3 and 3AO)with lipofectaminereagent.Control and vector groups were set up.Transfection and expression were confirmed by the means of RT-PCR.The biological characteristics of the transfected cellswith expression LPA receptor genes were studied by the observation of their growth and proliferative period.The effects of LPA on the of MMP-2 and MMP-9 protein expressions and activities in transfected ovarian cells were analyzed by gelatin zymography.Results It was confirmed that the LPA1 genes had been stably integrated into 3AO and SKOV3 cell lines.The growth characteristics of transfected cells were not different before and after transfectation.The high expression of LPA1 gene was revealed by the means of RT-PCR.The expression levels of LPA1 mRNA in transfected cells (SKOV3-LPA1 and 3AO-LPA1) were significantly higher than those in control transfected cells and an empty vector groups,there were significant differences (P<0.05).The activities and protein expressions of MMPs in the cells  transfected with LPA1 receptor genes did not increase,compared with control and empty vector groups (P>0.05).Conclusion The ovarian carcer cells with high expression of LPA1 gene in vitro have been established.LPA1 plays a role as negative growth receptor in the process of LPA-induced MMPs invasion and metastasis of ovarian carcer cells.

Key words: lysophosphatidic acid recepor;matrix metalloproteinase;ovarian neoplasms

中图分类号: 

  • R737.31