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目的:将溶血磷脂酸受体(LPA2、LPA3)基因转染卵巢癌细胞系SKOV3和3AO,探讨稳定表达LPA2和LPA3对细胞MMP-2和MMP-9活性的影响。方法:将pcDNA3.1flag-LPA2及pcDNA3.1flag-LPA3用脂质体介导转染SKOV3和3AO细胞,蛋白质印迹法鉴定细胞LPA2和LPA3蛋白表达。明胶酶谱法检测稳定表达LPA2和LPA3对卵巢癌细胞MMP-2和MMP-9表达及活性的影响。结果:LPA2和LPA3稳定转入SKOV3和3AO细胞,蛋白质印迹法分析结果证实,细胞内有LPA2和LPA3蛋白高表达。明胶酶谱分析结果显示,稳定表达LPA2和LPA3的基因可诱导SKOV3和3AO细胞MMP-2和MMP-9活性增加,与对照组及空载体组相比较,差异有统计学意义,P<0.05。结论:稳定表达LPA2和LPA3可诱导卵巢癌细胞MMPs活化,提示LPA2和LPA3在LPA诱导卵巢上皮性癌细胞MMPs侵袭和转移中可能发挥主要作用。
OBJECTIVE: To investigate the effect of LPA2 and LPA3 on the activity of MMP-2 and MMP-9 in ovarian cancer cell lines SKOV3 and 3AO by transfection of lysophosphatidic acid receptor (LPA2, LPA3) gene. Methods: pcDNA3.1flag-LPA2 and pcDNA3.1flag-LPA3 were transfected into SKOV3 and 3AO cells by lipofectamine 2000 respectively. The expression of LPA2 and LPA3 protein was detected by Western blotting. Gelatin zymography was used to detect the effect of LPA2 and LPA3 on the expression and activity of MMP-2 and MMP-9 in ovarian cancer cells. Results: LPA2 and LPA3 were stably transfected into SKOV3 and 3AO cells. Western blot analysis confirmed that LPA2 and LPA3 were highly expressed in the cells. The results of gelatin zymography showed that the stable expression of LPA2 and LPA3 can induce the increase of MMP-2 and MMP-9 activity in SKOV3 and 3AO cells, compared with the control group and empty vector group, the difference was statistically significant (P <0.05). Conclusion: Stable expression of LPA2 and LPA3 can induce MMPs activation in ovarian cancer cells, suggesting that LPA2 and LPA3 may play a major role in LPA-induced MMPs invasion and metastasis of ovarian epithelial cancer cells.