乙酰肝素酶反义寡核苷酸对肺癌A549细胞侵袭力抑制作用.docVIP

乙酰肝素酶反义寡核苷酸对肺癌A549细胞侵袭力抑制作用 　 作者：陈志涛 田辉 岳韦名 李林 亓磊 朱应超 【摘要】 目的 探讨乙酰肝素酶(heparanase，HPSE )反义寡核苷酸(antisense oligodeoxynucleotide，ASODN)对人肺癌A549细胞侵袭力的抑制作用。方法 设计合成互补于HPSE mRNA起始密码区的HPSE ASODN，以阳离子脂质体Lipofectin包埋后转染人肺癌A549细胞进行培养，采用流式细胞分析技术和逆转录聚合酶链反应(RTPCR)方法检测肺癌A549细胞HPSE蛋白及其mRNA表达的变化;以Matrigel细胞侵袭试验对HPSE ASODN对肺癌A549细胞侵袭的抑制进行检测。结果 ASODN各组与正常对照组和脂质体组比较及ASODN各组间比较HPSE mRNA和蛋白的表达及细胞侵袭力均受到明显抑制 (Plt;0.01);ASODN在终浓度为100、200、400 nmol/L时对肺癌A549细胞侵袭力的抑制率分别为55.6%、82.3%和91.2%。结论 HPSE ASODN通过下调HPSE mRNA和蛋白的表达可显著抑制肺癌A549细胞的侵袭力，并呈剂量依赖性。 【关键词】 肺肿瘤;乙酰肝素酶; 反义寡核苷酸 　　【Abstract】 Objective To evaluate the inhibitory effect of HPSE antisense oligodeoxynucleotide (ASODN) on the invasiveness of human lung cancer A549 cell lines.Methods HPSE ASODN which was complementary with initiation codon region of HPSE mRNA was designed and synthesized.After embedded by cation lipofectin，it was transfected into A549 cells of human lung cancer.The expression of HPSE protein and HPSE mRNA in A549 cell lines were detected by flow cytometry and RT PCR.Meanwhile Matrigel invasive assay was used to measure the inhibitory effect of HPSE ASODN on the invasiveness of human A549 cell lines.Results The HPSE protein and HPSE mRNA expression and invasiveness of human A549 cells treated with ASODN of different concentrations were significantly decreased as the ASODN concentration increasing.There was a significant difference between control group and each group of ASODN respectively(Plt;0.01)，between lipofectin group and each group of ASODN respectively(Plt;0.01)，and among the groups of ASODN (Plt;0.01).Besides，the difference of inhibitory effect was significant among the cells treated with different ASODN concentrations (Plt;0.01).The inhibition rates of A549 cell invasiveness were 55.6%，82.3 % and 91.2% treated by ASODN at final concentration of 100，200 and 400 nmol/L respectively.Conclusions By downregulating the expression of HPSE mRNA and HPSE protein，HPSE ASODN has significant inhibitory effect on