反义VEGF硫代寡脱氧核苷酸抑制裸鼠人肝癌移植瘤生长的研究

目的 :检测硫代修饰的反义脱氧寡核苷酸 (ASPODN)抑制人肝癌血管形成和生长的作用 ,以探讨ASPODN的临床应用前景。方法 :传代培养的肝癌细胞系HepG2 ,接种于裸鼠颈背部皮下 ,自 4 8h始局部注射ASPODN和等量的PBS ,彩色多普勒能量图 (CDE)检测移植瘤内新生血管的数目和分布 ;LSAB法免疫组化染色检测移植瘤中微血管密度 (MVD) ;FCM分析裸鼠移植瘤细胞周期动力学和凋亡率。结果 :局部注射ASPODN组和PBS对照组 ,裸鼠人肝癌移植瘤瘤重及MVD平均值分别为 ( 1.2 8±0 .2 6)g、2 6.90± 7.4 2和 ( 3.4 6± 0 .86)g、50 .36± 10 .2 7(P <0 .0 1) ;局部注射ASPODN组 ,用CDE动态检测到移植瘤内的血管数目及血流丰富程度明显低于PBS组P <0 .0 1;经ASPODN治疗后的裸鼠人肝癌移植瘤细胞凋亡率 ( 7.4 8± 1.64) %高于PBS对照组 ( 2 .75± 0 .2 2 ) % (P <0 .0 1)。结论 :VEGFAS PODN明量抑制肝癌血管形成和生长 ,有望成为理想的抗肝癌血管生成基因治疗的新药

Inhibition of Growth of Human Hepatocellular Carcinoma in Nude Mice Model by Antisense Phosophorothioate Oligodeoxynuclieotidesin

Objective To study the inhibition effect of antisense phosophorothioate oligodeoxynuclieotidesin(ASPODN) on growth and angiogensis of human hepatocellular carcinoma in nude mice model, investigatewhether ASPODN of VEGF can become kind of drug for antitumor gene therapy. Methods 5 × 106/ml HepG2were inoculated subcutaneously to 12 nude mices,they were randomized into 2 groups. From the 2 nd day, AS-PODN and PBS were given. The number and distribution of angiogenesis of sucutaneous tumors were detectedwith color doppler energy (CDE),microvesse density was detected with immunohistochemical stain. A cytoki-netic study and apoptosis rate of human hepatocellular carcinoma in nude mice model was performed by FCM.Results The tumor weight and MVD of ASPODN group were( 1.28 ± 0.26)g and 28.90 ± 7.42;PBS controlgroup were (3.46 ± 0.86) g and 50.36 ± 10.28, respectively ( P ＜ 0.01 ). The angiogenesis numers and distri-bution of human hepatocellular carcinoma in nude mice model in CDE was correlation with MVD ( P ＜ 0.05).Apotosis index in ASPODN group7.48 t 1.64 was higher than that in PBS group 2.75 ± 0.22 (P ＜ 0.01).Conciusions The growth and angiogenesis of human hepatocellular carcinoma in nude mice model can be in-hibited by ASPODN;VEGF ASPODN may become an new antitumor angiogenesisi gene therapy drugs.