环氧合酶-2反义寡核苷酸对宫颈癌Hela细胞的增殖抑制作用

The inhibition effection of Hela cell proliferation of cervical carcinoma by cyclooxygenase-2 antisense oligodeoxynucleotides

目的研究COX-2反义寡核苷酸（AS—ODNs）对宫颈癌Hela细胞的抑制作用。方法Hela细胞分以下6组培养：Hela细胞对照组（A），仅用常规培养液培养Opitin—MEM／MEM；正义组（B）：加入正义COX-2核苷酸至终浓度10μmol／1＋lipofectin＋Opitin—MEM；脂质体组（C）lipofectin＋Opitin—MEM。每组设4个复孔；反义寡核苷酸转染组（D）设置3个浓度亚组，分别转染AS—ODNs至终浓度为5μmol／1（D1）、10μmol／1（D2）、15μmol／1（D3）；在转染后24、48、72、96、120小时采用MTT方法检测各组细胞增殖状态，根据测得的OD值画细胞生长曲线。结果（1）与其他组相比，经AS—ODNs的细胞形态和生长状态都有明显改变，可见细胞生长状态差，增长缓慢该组细胞OD值与其它组比较，差异显著（P〈0．01）；（2）根据各组细胞的OD值，绘制细胞生长曲线，可见对照组、正义组、脂质体组生长曲线呈明显上升的趋势；AS—ODNs作用的三个亚组则呈下降趋势，且随浓度的增高，细胞生长曲线下降幅度增大，其抑制效果与作用时间有关，转染后48-72小时抑制效果最强，如果不行AS—ODNs补充，120小时抑制效果基本消失。结论COX-2反义寡核苷酸对宫颈癌Hela细胞的增殖具有抑制作用。

Objective To investigate the effect of the antisense oligodeoxynucleotides of human eyclooxygenase- 2 （COX-2 AS- ODNs） in inhibiting cervical cancer cell line Hela proliferation. Methods The Hela cells were cultured in vitro and were divided into 6 groups： control group （A）, cultured in MEM; Sense- ODNs group （B）, were cultured in COX- 2 10 μmol/l AS- ODNs ＋ lipofectin ＋ Opitin- MEM; Lipo group （C） （transfected with Lipofectin only）, D1 group （transfeoted with 5μmol/l COX- 2 AS- ODN＋ Lipo/ well）, D2 group （transfected with 10 μmol/l COX- 2 AS- ODN＋ Lipo/well）, and D3 group （transfected with 15μmol/l COX- 2 AS- ODN ＋ Lipo/well）. （1）The proliferative status of transfected cells was measured by using methabenzthiazuron （MTT） assay to detect the alive Hela calls in, 24,48,72,96 and 120 hours after the transfection, and cell proliferation curve according to the OD number. Results The lipoferation of the transfected Hela cells was significantly inhibited in the COX - 2 AS - ODN group, in the AS - ODNs transfected cells, the proliferative index decreased significantly with the increasing of COX - 2 AS - ODNs concentration. The effect of COX - 2 AS - ODN was strongest by the 48 ～ 72 hour after transfection and then began to decrease and basically disappeared 120 hours after, if no AS - ODNs is supplied. Conclusion Transfection with antisense ODNs of human COX - 2 mRNA could inhibit the proliferation of human cervical cancer Hela cells.