贝伐珠单抗联合高压氧治疗小鼠胶质母细胞瘤的效果

目的 探讨贝伐珠单抗（Bev）联合高压氧（HBO）对小鼠胶质母细胞瘤（GBM）的治疗作用。方法 体外培养U251细胞，随机分为对照组、Bev组（Bev作用24 h）、HBO组（HBO治疗1次）和Bev+HBO组（Bev作用24 h后，接受HBO治疗1次），采用MTT法检测细胞增殖能力，采用划痕实验和Transwell小室实验检测细胞迁移和侵袭能力。取100只雄性BALB/c裸鼠，右侧纹状体注射U251细胞建立GBM模型，造模后7 d随机分为对照组（腹腔注射PBS，3次/周，连续2周）、Bev组（腹腔注射Bev，5 mg/kg，3次/周，连续2周）、HBO组（HBO治疗，1次/d，连续2周）、Bev+HBO组（同时接受Bev和HBO处理2周）；每组取10只小鼠记录生存时间；每组取5只小鼠，HE染色测定肿瘤体积；每组取5只小鼠，CD34免疫组化染色测定肿瘤组织微血管密度（MVD）；每组取5只小鼠，PCR检测肿瘤组织基质金属蛋白酶（MMP）9 mRNA表达水平。结果 Bev对U251细胞增殖能力无明显影响（P>0.05），明显增强U251细胞迁移和侵袭能力（P<0.01）；明显降低肿瘤组织MVD（P<0.05），明显增加肿瘤组织MMP9 mRNA表达水平（P<0.05），但对荷瘤小鼠肿瘤体积及生存时间无明显影响（P>0.05）。HBO明显抑制U251细胞增殖、迁移、侵袭能力（P<0.01 ），明显降低荷瘤小鼠肿瘤组织MMP9 mRNA表达水平（P<0.01），但对荷瘤小鼠生存时间、肿瘤体积及肿瘤组织MVD均无明显影响（P>0.05）。Bev联合HBO明显抑制U251细胞增殖、侵袭和迁移能力，明显降低荷瘤小鼠肿瘤组织MVD和MMP9 mRNA水平（P<0.05），明显缩小肿瘤体积（P<0.05），明显延长荷瘤小鼠的生存时间（P<0.01）。结论 单独应用Bev或HBO对GBM小鼠肿瘤体积及生存时间无明显影响，但Bev联合HBO明显抑制小鼠GBM生长，明显延长小鼠生存时间。

Treatment of bevacizumab combined with hyperbaric oxygen therapy for a mouse model of glioblastoma

Objective To explore the therapeutic effect of bevacizumab (Bev) combined with hyperbaric oxygen therapy (HBO) on a mouse model of glioblastoma (GBM). Methods U251 cells were cultured in vitro and randomly divided into control group, Bev group (treatment with Bev for 24 h), HBO group (tretament with HBO for one time) and Bev+HBO group (treatment with Bev for 24 h and then with HBO). MTT assay, transwell migration assay and wound healing assay were used to detect the proliferation, migration and invasion abilities of U251 cells. U251 cells were injected into the right striatum of 100 male BALB/c nude mice to establish the GBM models. 7 days after making the establishment, these mice were randomly divided into control group (intraperitoneal injection of PBS, 3 times per week for 2 weeks) and Bev group (intraperitoneal injection of Bev, 5 mg/kg, 3 times per week for 2 weeks), HBO group (HBO, 1 time per day for 2 weeks), Bev+HBO group (treatment with Bev and HBO for 2 weeks). Ten mice in each group were used to record survival time; 5 mice in each group were used to measure the tumor volume by HE staining; 5 mice in each group were used measure the tumor tissue microvessel density (MVD) by CD34 immunohistochemical staining; 5 mice in each group were used to detect the expression level of matrix metalloproteinase (MMP) 9 mRNA in tumor tissues by PCR. Results Bev had no significant effect on the proliferation of U251 cells (P>0.05), but significantly enhanced the migration and invasion of U251 cells (P<0.01). Bev significantly reduced tumor tissue MVD (P<0.05) and significantly increased tumor tissue MMP9 mRNA expression levels (P<0.05), but had no significant effect on tumor volume and survival time of GBM mice (P>0.05). HBO significantly inhibited the proliferation, migration and invasion of U251 cells (P<0.01). HBO significantly reduced the expression level of MMP9 mRNA in tumor tissues of GBM mice (P<0.01), but it had no significant effect on the survival time, tumor volume and tumor tissue MVD of GBM mice (P>0.05). Bev combined with HBO significantly inhibited the proliferation, invasion and migration of U251 cells, significantly reduced MVD and MMP9 mRNA levels in tumor tissues and tumor volume of GBM mice (P<0.05), and significantly prolonged the survival time of GBM mice (P<0.01). Conclusions Bev or HBO alone has no significant effect on the tumor volume and survival time of GBM mice, but the combination of Bev and HBO significantly inhibits the tumor growth and significantly prolongs the survival time of GBM mice.