靶向抑制存活素表达对卵巢癌细胞株生物学行为的影响和意义

目的：通过存活素（ Survivin）反义寡核苷酸（ ASODN）转染人卵巢癌SKOV3细胞株抑制存活素Survivin蛋白表达，分析靶向抑制 Survivin 表达对卵巢癌细胞生长、细胞周期改变、凋亡、侵袭迁移能力的影响和意义。方法用脂质体（LipofectamineTM2000）介导Survivin ASODN转染人卵巢癌细胞株SKOV3，通过四唑盐比色法（MTT法）分析细胞生长活性改变，通过流式细胞仪检测Survivin蛋白表达改变、细胞周期分布和细胞凋亡率，通过Transwell小室检测细胞侵袭迁移能力的改变。结果转染Survivin ASODN后卵巢癌细胞株中Survivin蛋白表达明显下降（t＝7．82，P＜0．01）。细胞生长明显减慢（F＝3．75，P＜0．01），600nm/L ASODN转染48小时细胞存活率为49．50±20．76％，接近IC50值。细胞凋亡明显增加（t＝6．37，P＜0．05），细胞增殖明显下降（t＝-10．35，P＜0．01），更多的细胞停留在G0/G1期（t＝10．38，P＜0．01）。细胞迁移和侵袭能力下降（t值分别为19．26、27．42，均P＜0．01）。结论 Survivin ASODN通过靶向抑制卵巢癌细胞存活素表达，可抑制细胞生长，促进细胞凋亡，减少进入有丝分裂的细胞数，降低细胞的侵袭迁移能力，从而达到治疗卵巢癌的作用。靶向抑制存活素的治疗将成为卵巢癌治疗的重要手段。

Influence and significance of targeted inhibition of Survivin expression on biological behaviors of ovarian cancer cell line

Objective To study the influence and significance of targeted inhibition of Survivin on proliferation, apoptosis, migration and invasion of SKOV3 ceils by transfecting ovarian cancer cell line SKOV3 with antisense oligonucleotides aimed to Snrvivin （Survivin ASODN）. Methods Snrvivin ASODN was introduced by LipofectamineTM2000 to transfect reagent into SKOV3 cells, and growth activities of SKOV3 cells were detected by tetrazolium bromide （MTF） colorimetry. The changes of Survivn protein, cell cycle and apoptosis index （AI） were detected by flow cytometry （FCM）. The changes of invasion and migration were detected by Transwell. Results The protein level of Survivin was significantly down-regulated after transfection （ t =7.82, P 〈0.01 ）. Cell growth was significantly inhibited by various concentrations of ASODN （ F = 3.75,P 〈 0.01 ）. The cell survival rate was 49.50 ± 20.76% after 48 hours of transfection with 600 nmol/L ASODN, which was close to ICS0. AI increased significantly （ t = 6.37, P 〈 0.05 ） but PI decreased significantly （ t = - 10.35, P 〈 0.01 ）, and more cells stayed GI/G0 phase （t = 10. 38, P 〈0.01 ）. The migration ability and invasion ability of SKOV3 cells decreased （ t value was 19.26 and 27.42, respectively, both P 〈 0.01 ）. Conclusion Survivin ASODN could inhibit proliferation, promote apoptosis, decrease the proportion of mitotic ceils, reduce the migration ability and invasion ability and thus cure ovarian cancer. The targeted inhibition of Survivin will be an important therapy for ovarian cancer.