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目的:研究腺病毒介导的人ING4基因(Ad-ING4)对MG-63人骨肉瘤细胞移植瘤的生长抑制作用及其机制。方法:将本室构建好的重组腺病毒表达载体Ad-ING4,经QBI-293A细胞感染多轮扩增后获得高效价重组病毒子以用于肿瘤基因治疗试验,首先建立MG-63人骨肉瘤细胞移植瘤裸鼠模型,然后将15只荷瘤裸鼠随机分为阴性对照组(PBS组)、空载体对照组(Ad-GFP组)、Ad-ING4实验组(Ad-ING4组),3组均使用瘤体内注射干预用药,隔日一次,共5次。分别于用药前和用药后测量皮下瘤的体积,治疗开始后的第15天将裸鼠脱颈处死,摘取瘤体称重,计算抑瘤率。HE染色观察移植瘤细胞形态,免疫组化法检测瘤体中Bcl-2、Bax、Caspase-3、VEGF、CD34细胞因子的表达。结果:获得了高滴度(109pfu/ml)的重组腺病毒Ad-ING4;经瘤体基因治疗后,Ad-ING4组与PBS组及Ad-GFP组相比,可以明显抑制裸鼠MG-63人骨肉瘤细胞移植瘤生长,瘤重抑制率可达59.3%,移植瘤组织中可出现典型的细胞凋亡、坏死现象,其分子机制可能与Ad-ING4明显上调瘤体中促进凋亡的细胞因子Bax、Caspase-3的表达,下调抑制凋亡因子Bcl-2及血管形成细胞因子VEGF、CD34的表达有关。结论:Ad-ING4可以显著抑制MG-63人骨肉瘤细胞荷瘤生长,其机制可能通过激活细胞凋亡和抑制血管形成等途径来发挥抑瘤作用。
AIM: To investigate the effect of adenovirus-mediated human ING4 gene (Ad-ING4) on the growth of MG-63 human osteosarcoma xenografts and its mechanism. Methods: Ad-ING4, a recombinant adenovirus expression vector constructed in our laboratory, was obtained. After multiple rounds of infection with QBI-293A cells, a high titer recombinant virus was obtained for tumor gene therapy. First, MG-63 human osteosarcoma cells The nude mice were randomly divided into three groups: negative control group (PBS group), empty vector control group (Ad-GFP group), Ad-ING4 experimental group (Ad-ING4 group) All use of tumor injection therapy intervention, every other day, a total of 5 times. The volume of hypodermic tumor was measured before treatment and after treatment. On the 15th day after the start of treatment, the nude mice were sacrificed and the tumors were weighed and the tumor inhibition rate was calculated. The morphology of the transplanted tumor was observed by HE staining and the expression of Bcl-2, Bax, Caspase-3, VEGF and CD34 in the tumor was detected by immunohistochemistry. Results: The recombinant adenovirus Ad-ING4 with high titer (109pfu / ml) was obtained. Compared with PBS group and Ad-GFP group, Ad-ING4 group could significantly inhibit MG-63 Human osteosarcoma xenografts grew to a tumor weight of 59.3%. Typical apoptotic and necrotic phenomena were found in the xenograft tumors. The molecular mechanism may be related to Ad-ING4 upregulation of apoptosis-promoting cytokines Bax, Caspase-3 expression, downregulation of apoptosis-inhibiting factor Bcl-2 and angiogenic factors VEGF, CD34 expression. Conclusion: Ad-ING4 can significantly inhibit MG-63 human osteosarcoma cell tumor growth, its mechanism may play an antitumor effect by activating apoptosis and inhibiting angiogenesis.