摘要
目的:研究溶血磷脂酸受体(LPA1、LPA2、LPA3)的反义寡核苷酸(ASODN)对卵巢癌细胞的抑制作用;探讨LPA对表达LPA受体ASODN的卵巢癌细胞株MMP2和MMP9表达及活性的影响。方法:将LPA1、LPA2和LPA3ASODN转入卵巢癌细胞系SKOV3和3AO,用RT-PCR和蛋白质印迹法鉴定转染细胞株的表达水平。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,明胶酶谱检测分析LPA对表达LPA受体的ASODN的卵巢癌细胞株MMP-2、MMP-9表达及活性的影响。结果:LPA1、LPA2及LPA3ASODN均可有效的抑制LPA1基因、LPA2及LPA3蛋白质的表达。转染前后细胞株生长情况差异无统计学意义。明胶酶谱条带的灰度值分析显示,LPA诱导表达LPA2、LPA3ASODN的卵巢癌细胞株MMP-2、MMP-9活性降低,与对照组及正义组相比,差异有统计学意义,P<0.05。转染LPA1ASODN的卵巢癌细胞株MMP-2、MMP-9活性,与对照组及正义组相比,差异无统计学意义,P>0.05。结论:ASODN抑制LPA受体基因及蛋白质表达。在LPA诱导的MMPs活化过程中,LPA2及LPA3更有效,提示卵巢上皮性癌中LPA2和LPA3在LPA诱导的MMPs侵袭、转移过程中发挥主要作用。
OBJECTIVE：To evaluate the influence of LPA1,LPA2 and LPA3 antisense oligodeoxynucleotides（ASODN） on inhibition of human ovarian cancer 3AO and SKOV3 cells,and study the effects of lysophosphatidic acid （LPA） on MMP-2 and MMP-9 expressions and activities in ovarian cancer cells with transfected LPA receptors ASODN. METHODS：The LPA1,LPA2 and LPA3 antisense oligodeoxynucleotides （ASODN） were integrated into 3AO and SKOV3 cells lines. Transfection and expression were confirmed by the means of RT-PCR and Western blotting. The biological characteristics of the transfected cells were studied by the observation of their growth and proliferative periods. The effect of LPA on MMP-2 and MMP-9 protein expressions and activities in transfected ovarian cells was analyzed by gelatin zymography. RESULTS：The LPA1 gene,LPA2 and LPA3 protein expressions were decreased in ovarian cancer cells with LPA1,LPA2 and LPA3 ASODN transfection. The transfected cells growth characteristics were not different before and after the transfectation. The gelatin zymography confirmed that LPA-induced the activities and protein expressions of MMP-2 and MMP-9 decreased in the transfected cells with LPA2 and LPA3 receptors ASODN,compared with those in the control group and SODN group （P〈0.05）. The activities and protein expressions of MMPs in LPA1 receptors ASODN cells did not decrease,compared with those in the control group and SODN group （P〉0.05）. CONCLSUIONS：LPA receptors antisense oligodeoxynucleotides can inhibite the expressions of LPA receptors gene and protein. LPA2 and LPA3 were more efficient in LPA-induced MMPs activity. LPA2 and LPA3 play important roles in the process of LPA-induced MMP invasion and metastasis of ovarian carcer cells.
出处
《中华肿瘤防治杂志》
CAS
2010年第1期50-54,共5页
Chinese Journal of Cancer Prevention and Treatment
关键词
卵巢肿瘤
溶血磷脂酸
受体
基质金属蛋白酶类
ovarian neoplasms
lysophosphatidic acid
receptors
matrix metalloproteinases
