FAP-1分子对卵巢癌细胞株SKOV3凋亡的影响

Effects of FAP-1 on apoptosis of ovarian cancer cell line SKOV3

目的探讨Fas相关磷酸酯酶-1(FAP-1)对卵巢癌细胞株SKOV3凋亡的影响.方法采用免疫印记法(Western blot)和逆转录聚合酶链式反应(RT-PCR)方法,检测了经FAP-1反义寡核苷酸(FAP-1ASODN)封闭前后FAP-1蛋白与核酸在卵巢癌细胞株SKOV3中的表达水平;应用四唑盐比色法(MTT法)和流式细胞术(FCM)检测了经抗Fas激活性抗体DX2诱导凋亡后FAP-1分子对SKOV3细胞凋亡行为的影响.结果FAP-1蛋白与mRNA在卵巢癌细胞株SKOV3中均有明显表达.当用DX2以不同时间和不同浓度诱导凋亡后,DX2+FAP-1ASODN组在12、24、48和72 h的细胞抑制率约为DX2组的1～2倍,细胞凋亡率约为后者的2～4倍,差异有显著性(P＜0.05,P＜0.01);在DX2低、中、高3个浓度时,DX2+FAP-1ASODN组的细胞抑制率为(20.94±1.15)%、(34.70±1.24)%、(45.45±2.80)%,DX2组分别为(11.42±0.38)%、(18.33±1.29)%、(28.75±1.81)%,差异有显著性(P＜0.05,P＜0.01);而在DX2组与DX2+FAP-1SODN组间无显著性差异(P＞0.05).结论FAP-1分子在卵巢癌细胞SKOV3中有明显表达,且具有抵抗Fas介导凋亡的功能,而FAP-1ASODN可以明显提高卵巢癌细胞对凋亡的敏感性,可能对卵巢癌的发病与治疗具有重要意义.

Objective To investigate the effects of Fas associated phosphatase-1(FAP-1)on apoptosis of ovarian cancer cell line SKOV3. Methods Western blot and reverse transcriptase-polymerase chain reaction(RT-PCR) analysis were employed to detect the expression of FAP-1 in cell line SKOV3 with or without pre-treatment with FAP-1 antisense oligonucleotide (FAP-1ASODN), and MTT method and flow cytometry were used to evaluate the apoptosis of cell line SKOV3 induced by agonistic anti-Fas antibody (DX2). Results There was evident expression of FAP-1 protein and mRNA in cell line SKOV3. When induced apoptosis by DX2 for 12-72 hrs, the cell inhibitive rates of “DX2+FAP-1ASODN” group were 1-2 times as those of “DX2” group ( P <0.05 or P <0.01), and the apoptotic rates were 2-4 times as those of “DX2” group ( P <0.05 or P <0.01). Moreover, when treated with DX2 by low, moderate and high doses, the cell inhibitive rates were (20.94±1.15)%, (34.70± 1.24)% and (45.45±2.80)% in “DX2+FAP-1ASODN” group vs. (11.42±0.38)%, (18.33±1.29)% and (28.75± 1.81)% in “DX2” group ( P <0.05 or P <0.01) respectively. But there was no significant difference between “DX2” group and “DX2+FAP-1SODN” group ( P >0.05). Conclusion FAP-1 is overexpressed in cell line SKOV3 and it may help the cell resist Fas-mediated apoptosis, while FAP-1ASODN can increase the Fas-sensitivity of this ovarian cancer cell. So it seems that FAP-1 may be involved in the pathogenesis of ovarian cancer.