FAP-1反义寡核苷酸联合卡铂对卵巢癌细胞SKOV3凋亡的作用

Combinative Effects of FAP-1 Antisense Oligonucleotide and Carboplatin on Apoptosis of Ovarian Cancer Cell SKOV3

背景与目的实验已证明,Fas相关磷酸酯酶-1(FAP-1)在卵巢癌细胞SKOV3中有高表达,可能对卵巢癌的发病与耐药具有重要意义。本研究应用反义核酸技术探讨FAP-1反义寡核苷酸(FAP-1ASODN)联合卡铂对SKOV3细胞凋亡的作用。方法反义核苷酸技术将FAP-1ASODN转染SKOV3细胞。采用逆转录聚合酶链反应(reversetranscription-polymerasechainreaction,RT-PCR)检测转染前后FAP-1mRNA的表达,采用流式细胞术(FCM)分析细胞周期与细胞凋亡率,噻唑蓝法(MTT法)测定FAP-1ASODN与40μg/ml卡铂共同作用后对SKOV3细胞生长的影响。结果经FAP-1ASODN处理24h后,与对照组及FAP-1正义寡核苷酸(SODN)组比较,反义组的FAP-1mRNA表达明显减弱。FCM结果显示,卡铂+FAP-1ASODN组在24～72h的细胞凋亡率明显高于单用ASODN组及单用卡铂组,差异有显著性(P<0.05),且可将细胞周期阻滞于G1期;MTT结果显示,卡铂+FAP-1ASODN组12～96h的细胞抑制率约为单用ASODN组的1.5～2倍,约为单用卡铂组的1.5倍,明显高于后两者,差异有显著性(P<0.05);而在单用卡铂组与卡铂+FAP-1SODN组间无显著性差异(P>0.05)。结论FAP-1ASODN转染可以有效抑制FAP-1基因的表达,同时可以增强卵巢癌细胞对卡铂诱导凋亡的敏感性。

BACKGROUND &OBJECTIVE: Recent studies have shown that overexpression of Fas associated phosphatase 1 (FAP 1) can been detected in human ovarian cancer cell line SKOV3, suggesting that this overexpression may play an important role in the tumorigenesis and drug resistance of ovarian cancer. This study was designed to explore the effects of fas associated phosphatase 1 antisense oligonucleotide (FAP 1 ASODN) combined with carboplatin on the apoptosis of ovarian cancer cell SKOV3. METHODS: Antisense oligonucleotide technique was used to transfect FAP 1 ASODN into SKOV3 cells. The expression levels of FAP 1 mRNA of SKOV3 cells with or without FAP 1 ASODN transfection were determined by reverse transcription polymerase chain reaction (RT PCR). The cell cycle and apoptotic rate were analyzed by flow cytometry (FCM). The growth and proliferation of SKOV3 cells were observed by MTT assay. RESULTS: After transfected FAP 1 ASODN for 24 hours, the expression of FAP 1 mRNA in SKOV3 cells was obviously reduced compared with those of the control and FAP 1 SODN transfection groups. When induced apoptosis with 40 μg/ml carboplatin for 24-72 hours, FCM results showed the apoptotic rate of “carboplatin+FAP 1 ASODN”group was higher than those of “carboplatin”group and “ASODN”group (P< 0.05), and the cell cycle was blocked in G1 phase. MTT results showed the cell inhibitory rate of “carboplatin+FAP 1 ASODN”group was 1.5-2 times those of “carboplatin”group and “ASODN”group (P< 0.05); but no significant difference was found between “carboplatin”group and “carboplatin+FAP 1 SODN”group (P >0.05). CONCLUSION: FAP 1 ASODN transfection can suppress the expression of FAP 1 gene in SKOV3 cells and enhance the cell sensitivity to apoptosis induced by carboplatin, which implies that FAP 1 ASODN may reverse the drug resistance in ovarian cancer.