乙酰肝素酶反义寡核苷酸对Hep-2细胞侵袭力抑制的研究.pdfVIP

中国耳鼻咽喉头颈外科／2008年2月，第1 5卷，第2期 乙酰肝素酶反义寡核苷酸对Hep一2细胞侵袭力抑制的研究 马建刚’，李晓明 ，路秀英 (1河北医科大学第二医院耳鼻咽喉科，河北 石家庄 050000：2白求恩国际和平医院耳鼻咽喉头颈外科，河北 石家庄 050082) 【摘 要】 目的 探讨乙酰肝素酶 (heparitinase， inhibitory ef1．ect 0f HPSE AS．ODN on the invasiveness HPSE)反义寡核苷酸 (antisense 0Iig0dexyrIb0nucIe· of human Hep-2 cell Iines．METHODS HPSE AS． otides，AS·0DN)对Hep-2细胞HPSE mRNA、HPSE ODN which was complementary with initiation codon 蛋白的表达及其侵袭力的活性抑制。方法 设计合成 region of HPSE mRNA was designed and synthesized． 互补于HPSE mRNA起始密码区的HPSE AS·ODN，以阳 After embedded by cation Iipofectin．it was transfected 离子脂质体Lipofectin包埋后转染人喉癌Hep-2细胞进行 into HeP-2 cells of human Iaryngocarcinoma．The 培养，采用流式细胞分析技术和逆转录-聚合酶链反应 expression of HPSE protein and HPSE mRNA in Hep-2 (reverse transcription-polymerase chain reaction，RT． celllines were detected by flow cytometry and RT． PCR)方法检测Hep-2细胞HPSE蛋白及其mRNA表达的 PCR．Meanwhile MatrigeIinvasive assay was used to 变化；以Matrigel_~1]]胞侵袭试验分别对HPSE AS-ODN对 measure the inhibitory effect of HPSE AS．ODN on the Hep-2细胞侵袭力的抑制进行检测。结果 AS-ODN各 invasiveness of human Hep．2 cell Iines．RESULTS 组与正常对照组和脂质体组比较及AS-ODN各组间比较 The HPSE protein and HPSE mRNA expression and HPSE mRNA和HPSE蛋白的表达及细胞侵袭力均有明 invasiveness of human Hep．2 cells treated with AS- 显抑制作用，差异有统计学意义 (PO．01)：AS·ODN ODN 0f di仟erent c0ncentrati0ns were significantly 在终浓度为1 00、200、400 nmol／L时对Hep-2~1]]胞侵袭 decreased as the AS．ODN c0ncentrati0n increasing． 力的抑制率分别为60．1％、8O．8％and 94．0％。结论 There was a significantly di仟erence between controI HPSEASON通过下调HPSEmRNA和HPSE蛋白的表达 group and each group of AS-ODN respectively 可显著抑制Hep-2细胞的侵袭力并呈剂量依赖性。 (PO．01)，between lipofectin group and each group 【关键词】肝素裂合酶：寡脱氧核糖核苷酸类，反义； 0f AS·ODN respectively(PO．01)，and among the 喉肿瘤：肿瘤侵袭 groups of AS-ODN(PO．01)．Besides，the difference