肿瘤基因治疗学研究:血管内皮生长因子反义寡核苷酸对小鼠Lewis肺癌生长的抑制

Inhibitory effect of vascular endothelial growth factor antisense oligonucleot ide on the growth of Lewis lung cancer:a study on gene therapeutics for tumors

目的:探讨血管内皮生长因子(VEGF)反义寡核苷酸(ASPODN)治疗肺癌的可能性。方法:实验于2002-12/2004-05在哈尔滨医科大学第一临床医学院中心实验室完成。制备C57BL/6小鼠皮下肺癌模型30只,随机分为3组(每组10只):VEGF-ASPODN治疗组,VEGF正义寡核苷酸(SPODN)治疗组及对照组。接种Lewis肺癌细胞后24h内,分别皮下注射AS-PODN及SPODN进行治疗,对照组只注射生理盐水,2次/周,连续4周;观察各组小鼠肿瘤的生长情况、游标卡尺测量肿瘤体积大小。断颈处死小鼠,光镜及电镜下观察肿瘤组织形态学改变及超微结构变化。结果:与对照组瘤质量犤(7.83±0.78)g犦比较,VEGF-ASPODN组犤(4.49±0.43)g犦能明显抑制小鼠肿瘤生长(P<0.01),VEGF-SPODN组犤(7.73±0.69)g犦则无明显作用(P>0.05)。VEGF-ASPODN组和VEGF-SPODN组抑瘤率分别为42.7%和5.9%。组织形态学及超微结构观察,VEGF-ASPODN能明显抑制肿瘤细胞生长,降低增殖活性。结论:肿瘤原位注射VEGF-ASPODN能抑制小鼠肺癌生长。

AIM:To study the possibility of the therapy of vascular endothelial growth fac tor(VEGF) antisense oligonucleotide(ASPODN) for lung cancer. METHODS:The experiment was conducted in the central laboratory of the First Cl inical Medical College of Harbin Medical University from December 2002 to May 20 04.The Lewis lung cancer cells were cultured and implanted subcutaneously into 3 0 C57BL/6 mice,and subsequently the mice were divided into three groups with 10 mice in each group:VEGF ASPODN group,VEGF sense oligonucleotide(SPODN) group,a nd the control group.Subcutaneous injection of ASPODN plus normal saline(NS),SPO DN plus NS and NS were given to VEGF ASPODN group,VEGF SPODN group and control group respectively 24 hours after 4 week cell inoculation.The growth status of tumors in all the mice was observed,and volume of subcutaneous tumors was measu red with vernier caliper.After the mice were scarified by incising their necks,t he morphological and ultrastructural changes of tumor cells were observed under light microscope and electron microscope. RESULTS:As compared with the tumor mass of the control group[(7.83±0.78) g],t hat of ASVEGF ASPODN group[(4.49±0.43) g]was lower,so the tumor growth was inhi bited significantly(P< 0.01),but tumor mass of VEGF SPODN group was (7.73±0.69 ) g,indicating the tumor growth was not inhibited(P >0.05).The inhibition rate o f tumor growth in the VEGF ASPODN group and VEGF SPODN group was 42.7%and 5.9 %respectively.The observation of morphological and ultrastructural changes show ed that VEGF ASPODN could obviously inhibit the growth of tumor cells and decel erate the proliferation of tumor cells. CONCLUSION:Lewis lung cancer can be inhibited by the in situ injection of VEGF ASPODN into the tumor of C57BL/6 mice.