COX2反义核酸对喉癌恶性生物学行为的抑制作用.doc

　　COX2反义核酸对喉癌恶性生物学行为的抑制作用 ：姚小宝，王晓侠，张少强，闫利英，朱宏亮 【摘要】 目的 探讨环氧合酶2(COX2)反义核酸对喉癌细胞恶性表型的抑制作用。方法 采用脂质体介导方法，分别构建转染COX2反义真核表达载体和空载体的喉癌细胞系Hep2AS和Hep2P细胞。RTPCR及TT比色实验、Boyden侵袭小室法和裸鼠成瘤实验分别检测转染细胞的体外增殖速度、体外侵袭力和体内成瘤性。结果 RTPCR结果显示，Hep2AS细胞COX2/磷酸甘油醛脱氢酶(GAPDH)mRNA比值(0.65)明显低于Hep2(0.92)和Hep2P细胞(0.90)；TT比色实验显示Hep2AS细胞较Hep2P、Hep2细胞生长速度减慢，三者倍增时间分别为3.6d、2.9d和2.9d；体外侵袭实验结果表明，Hep2AS侵袭细胞数(18.20±5.97)显著低于Hep2细胞 (45.40±8.12)及Hep2P细胞(44.10±6.47)(Plt;0.01)。裸鼠成瘤实验中，Hep2P、Hep2细胞接种裸鼠后第5天肿瘤长出，而Hep2AS细胞第7天肿瘤长出；28d后Hep2AS细胞接种组瘤体体积[(764.85±129.61)mm3]低于Hep2细胞[(1181.73±360.83)mm3]和Hep2P细胞接种组[(1065.21±237.46)mm3]，有显著性差异(Plt;0.01)。结论 喉癌细胞中COX2过表达与细胞的恶性表型相关，反义核酸技术抑制COX2表达可以逆转喉癌细胞的恶性表型。 【关键词】 喉癌；核酸；环氧合酶2 ABSTRACT: Objective To explore the effect of antisense nucleic acid of cyclooxygenase2 (COX2) in malignant phenotype of laryngeal cancer cells and the mechanism of COX2 in carcinogenesis of laryngeal cancer. Methods Using LipofectamineTM2000 reagent, the COX2 highly expressed in human laryngeal cancer cell line Hep2 id pcDNA3.1 (named as Hep2AS and Hep2P cells, respectively). Semiquantitative RTPCR and TT assay, Boyden chamber and tumor implantation experiment origenesis of the transfected cells in vitro and in vivo. Results RTPCR analysis indicated that the ratio of COX2 mRNA to GAPDH mRNA in Hep2AS cells (0.65) ore sloe of 3.6, 2.9 and 2.9d, respectively. Boyden chamber assay revealed that the cell number of the Hep2AS cells (18.20±5.97) that migrated through the filter aller than that of the Hep2 (45.40±8.12) and Hep2P cells (44.10±6.47)(Plt;0.01). The tumor xenografts occurred on d5 in Hep2 and Hep2P cells, but on d7 in Hep2AS cells. On d30 after implantation, the volume of Hep2AS cell tumor xenografts [(764.85±129.61)mm3] m3] and Hep2P cells [(1065.21±237.46)mm3] (Plt;0.01). Conclusion Overexpression of COX2 in human laryngeal cancer cell line Hep2 is associated alignant phenotype of cancer cells. Inhibition of COX2 express