Experimental Study of Efficacy of Multi-target Inhibitor Combined with Cytotoxic Treatment on Malignant Glioma
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摘要: 目的 研究多靶点抑制剂凡德他尼联合卡莫司汀(BCNU)对C6鼠胶质瘤增殖及侵袭的影响。 方法 传代培养C6大鼠胶质瘤细胞， MTT比色法检测药物对细胞活性的影响，HE染色观察细胞形态的变化。原位注射法建立C6大鼠胶质瘤模型，6天后分为凡德他尼组、BCNU组、联合药物组（凡德他尼+BCNU）及空白组，隔天给药7次，两周后断头法处死大鼠取脑，计算肿瘤体积，做肿瘤组织切片；SP法标记肿瘤血管内皮细胞,检测移植瘤中微血管密度（MVD）；检测与肿瘤侵袭性有关的MMP1蛋白表达；TUNEL法检测肿瘤细胞凋亡指数。 结果 联合药物组促进肿瘤细胞凋亡效果显著高于BCNU组（P<0.01)。与空白组比较，BCNU组移植瘤体积下降了34.0％(P<0.05)，移植瘤中MMP1表达为(+〖KG-*2〗+)，MVD无明显变化，细胞凋亡率为62.6％(P<0.05)；联合药物组移植瘤体积下降了56.0％(P<0.05)，移植瘤中MMP1表达阳性(+)，MVD减少了53％(P<0.05），细胞凋亡率为82.0％(P<0.05)。 结论 联合用药不仅能明显抑制C6大鼠胶质瘤细胞移植瘤生长，还能降低与肿瘤侵袭能力有关的MMP1蛋白表达，这为临床上多靶点抑制剂与细胞毒药物联合治疗恶性胶质瘤提供了实验依据。Abstract: Objective To investigate the influence of multi-target inhibitor vandetanib combined cytotoxic drugs carmustine (BCNU) on the proliferation and invasion of rat glioma cell C6. Methods The MTT assay was used to test the impact of drugs on cell viability and changes of cell morphology were observed by HE staining. Rat glioma cell C6 model was established by situ injection, and six days later they were divided into the vandetanib group [50 mg/(kg·d)], BCNU group [25 mg/(kg·d)], the combination group (vandetanib+ BCNU) and the control group. Dose the rats 7 times every other day for 2 weeks, then take the rats' brain after the decapitation, section the tumor tissues and calculate the volume of the tumor. Immunohistochemical SP assay was used to mark tumor vascular endothelial cells, and detect the microvessel density (MVD) in transplanted tumors and the expression of MMP1 protein related to tumor invasiveness. Apoptotic index was tested by TUNEL technique. Results The combination group was much remarkable to promote the apoptosis than that of the BCNU group (P <0.01). Compared with the control group, tumor volumes of the BCNU group decreased by (P<0.05) The expression of MMP1 in xenografts was weak positive(++), and there was no significant change in MVD，but the apoptosis rate of transplanted tumor cells was 62.6%(P<0.05). Tumor volume of the combination group wasdecreased about 56% and the expression of MMP1 was negative(+), and MVD decreased by 53%(P <0.05). Conclusion The combination therapy can not only significantly inhibit the growth of glioma cell C6, but also decrease the expression of MMP1 related to neoplasm invasiveness. Therefore, it provides experimental basis for the joint multi-target inhibitors and cytotoxic drugs in clinical treatment of malignant glioma.
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Key words:
- C6 rat glioma /
- Multi-target inhibitors /
- Cytotoxic drugs /
- Efficacy studies
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