Wang Bin, Zheng Wei-Guo, Xin Xiao-Yan, Qi Rong-Yi, Yu Yue-Cheng, Cao Yun-Xin
Department of Obstetrics and Gynecology, 254 Hospital, PLA, Tianjin, 300142, PR China.
Ai Zheng. 2004 Aug;23(8):885-9.
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.
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.
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 microg/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).
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.
近期研究显示,在人卵巢癌细胞系SKOV3中可检测到Fas相关磷酸酶-1(FAP-1)的过表达,提示这种过表达可能在卵巢癌的发生及耐药中起重要作用。本研究旨在探讨Fas相关磷酸酶-1反义寡核苷酸(FAP-1 ASODN)联合卡铂对卵巢癌细胞SKOV3凋亡的影响。
采用反义寡核苷酸技术将FAP-1 ASODN转染至SKOV3细胞。通过逆转录-聚合酶链反应(RT-PCR)检测转染或未转染FAP-1 ASODN的SKOV3细胞中FAP-1 mRNA的表达水平。采用流式细胞术(FCM)分析细胞周期及凋亡率。通过MTT法观察SKOV3细胞的生长和增殖情况。
转染FAP-1 ASODN 24小时后,与对照组和FAP-1 SODN转染组相比,SKOV3细胞中FAP-1 mRNA的表达明显降低。用40μg/ml卡铂诱导凋亡24 - 72小时后,FCM结果显示“卡铂+FAP-1 ASODN”组的凋亡率高于“卡铂”组和“ASODN”组(P<0.05),细胞周期阻滞于G1期。MTT结果显示“卡铂+FAP-1 ASODN”组的细胞抑制率是“卡铂”组和“ASODN”组的1.5 - 2倍(P<0.05);但“卡铂”组与“卡铂+FAP-1 SODN”组之间无显著差异(P>0.05)。
FAP-1 ASODN转染可抑制SKOV3细胞中FAP-1基因的表达,并增强细胞对卡铂诱导凋亡的敏感性,这提示FAP-1 ASODN可能逆转卵巢癌的耐药性。