论文部分内容阅读
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 study the inhibitory effect of antisense oligonucleotide (ASODN) of lysophosphatidic acid receptor (LPA1, LPA2, LPA3) on ovarian cancer cells and to investigate the effect of LPA on the expression of MMP2 and MMP9 in ovarian cancer cell lines expressing LPA receptor ASODN And the impact of activity. Methods: LPA1, LPA2 and LPA3ASODN were transfected into ovarian cancer cell lines SKOV3 and 3AO, and the expression levels of the transfected cell lines were identified by RT-PCR and Western blotting. The biological characteristics of transfected cell lines were observed by detecting the growth and proliferation of cell lines. The effect of LPA on the expression and activity of MMP-2 and MMP-9 in ovarian cancer cell line ASODN expressing LPA receptor was analyzed by gelatin zymography. Results: LPA1, LPA2 and LPA3ASODN could effectively inhibit the expression of LPA1, LPA2 and LPA3 protein. There was no significant difference in cell growth between before and after transfection. The results of gray value analysis of gelatin zymogram showed that the activity of MMP-2 and MMP-9 in ovarian cancer cells LPA2 and LPA3ASODN induced by LPA decreased compared with the control group and the sense group, P < 0.05. The activity of MMP-2 and MMP-9 in ovarian cancer cell line transfected with LPA1ASODN was not significantly different from that in control group and normal group (P> 0.05). Conclusion: ASODN inhibits LPA receptor gene and protein expression. In LPA-induced activation of MMPs, LPA2 and LPA3 are more effective, suggesting that LPA2 and LPA3 in epithelial ovarian cancer play an important role in the invasion and metastasis of LPA-induced MMPs.