摘要
目的:利用血管内皮细胞瘤来源的细胞系EOMA来探讨Avastin治疗小鼠血管内皮细胞瘤的可行性。方法:利用CCK-8试剂检测不同质量浓度的Avastin(0、50、100、200 mg/L)对EOMA细胞增殖的影响。将EOMA细胞移植裸鼠制作动物模型,待肿瘤体积生长至100 mm3左右时,将动物随机分为2组,采取瘤内注射的方式给药,给药组按1 mg/kg体重给予Avastin,对照组给予等体积的磷酸盐缓冲液(PBS),每周给药2次,同时每周测量肿瘤体积及小鼠体重3次。实验结束时将小鼠处死,剥取肿瘤制作石蜡切片,免疫组织化学染色检测增殖细胞核抗原(PCNA),采用原位末端标记法TUNEL检测细胞凋亡,观察Avastin对肿瘤细胞增殖和凋亡的影响。小鼠处死前,在深度麻醉状态下行心脏穿刺取血收集血清,利用酶联免疫吸附实验检测各组小鼠血清中血管内皮细胞生长因子(VEGF)的表达水平。结果:体外实验显示,Avastin能够抑制EOMA细胞的增殖,不同浓度Avastin的抑制率分别为24.21%、26.26%和34.58%。实验结束时,PBS对照组小鼠的肿瘤体积为(1 860.10±146.96)mm3,而Avastin给药组的肿瘤体积为(681.45±63.01)mm3(P<0.01),提示瘤内注射Avastin能明显抑制肿瘤的生长。免疫组织化学PCNA染色结果显示,Avastin可以抑制细胞增殖(P<0.05);TUNEL染色提示Avastin可以明显诱导细胞凋亡增加(P<0.01)。Avastin给药组血清中VEGF的水平为(594.65±118.79)ng/L,明显低于PBS对照组的(802.24±238.41)ng/L(P<0.05)。结论:实验结果提示,Avastin可以作为一种治疗血管内皮细胞瘤的安全有效的药物,或许可以考虑将其应用于其他血管瘤样病变的治疗。
Objective： Investigated the efficacy of Avastin on murine hemangioendothelioma in vitro and in vivo using a mouse hemangioendothelioma-derived cell line-EOMA. Methods： The in vitro effect of Avastin on cell proliferation of EOMA cell line was measured by CCK-8 assay at 0, 50, 100, 200 mg/L concentration of Avastin. When tumors produced by subcutaneous injection of EOMA cells reached a volume of 100 mm^3, animals were treated by intratumor injection with Avastin （ 1 mg/kg body weight） or with vehicle alone （PBS） twice a week. Mice were weighted and tumors were measured three times weekly. At the end of the experiment, the mice were sacrificed and their tumors were excised and processed for histology. Immunohistochemieal study of apoptosis was conducted using a TUNEL kit, tumor cell proliferation was assessed with anti-proliferating cell nueler antiger （PCNA） antibody. Cardiac puncture was performed under deep anesthesia for collection of serum, serum vascular endothelial growth factor （VEGF） level was tested by VEGF-ELISA assay. Results： Avastin exhibited cell inhibited rate of 24. 21%, 26. 26% and 34. 58% at 50, 100 and 200 mg/L, respectively. When experiment was terminated, the tumor volume in the PBS-treated mice was （ 1 860. 10 ±146. 96） mm^3, being significantly larger than that in the mice that were treated by Avastin [ （681.45 ±63. 01 ） mm^3, P 〈0. 01 3. Avastintreated tumors showed decreased tumor cell proliferation and increased cell apoptosis. The VEGF level in mice treated with Avastin [ （594. 65± 118.79 ） ng/L] was significantly lower than that in PBS-treated mice [ （802.24± 238.41 ） ng/L, P 〈 0. 05 ]. Conclusion： The results suggest that topically applied Avastin provides an effective and safe approach to treat hemangioendotheliomas and might be used as a novel treatment of angiomatous diseases in the future.
出处
《北京大学学报（医学版）》
CAS
CSCD
北大核心
2009年第1期105-108,共4页
Journal of Peking University:Health Sciences
基金
国家自然科学基金(30471895)资助~~
关键词
血管内皮瘤
抗肿瘤药
内皮生长因子
内皮
血管
小鼠
Hemangioendothelioma
Antineoplastic agents
Endothelial growth factors
Endothelium, vascular
Mice
