论文部分内容阅读
目的 :探讨毒蕈碱型胆碱受体3(muscarinic cholinergic receptor 3,M3R)拮抗剂4-二苯乙酰氧基-N-甲基-哌啶甲碘化物(4-diphenylacetoxy-Nmethylpiperidine methiodide,4-DAMP)对裸鼠体内小细胞肺癌生长和血管生成的影响。方法:RT-PCR及蛋白质印迹法检测人小细胞肺癌细胞株SBC3中M3R mRNA及蛋白的表达。建立人小细胞肺癌SBC3细胞的裸鼠移植瘤模型,随机分为4组,分别腹腔注射0.9%氯化钠溶液(对照组)和不同剂量(0.5、1和2 mg/kg)的4-DAMP,每天给药1次,连续15 d。每2 d测量1次肿瘤体积;给药结束后处死裸鼠,测量肿瘤质量并计算抑瘤率。实时荧光定量PCR和蛋白质印迹法分别检测各组肿瘤组织中M3R和血管内皮生长因子(vascular endothelial growth factor,VEGF)mRNA和蛋白的表达水平。免疫组织化学法检测VEGF表达和微血管密度(microvessel density,MVD)。结果:人小细胞肺癌SBC3细胞表达M3R mRNA及其蛋白。成功建立人SBC3细胞的裸鼠移植瘤模型,并用不同剂量(0.5~2 mg/kg)的4-DAMP给药后,发现裸鼠体内移植瘤的体积均比对照组明显减小(P值均<0.05),剥离肿瘤的质量也明显减少(P值均<0.01),0.5、1和2 mg/kg 4-DAMP给药组的抑瘤率分别为15.82%、33.50%和55.06%。0.5~2 mg/kg 4-DAMP给药组肿瘤组织中M3R和VEGF的mRNA及蛋白表达水平均明显下调(P值均<0.05),MVD也明显下调(P值均<0.05)。结论 :M3R拮抗剂4-DAMP可以抑制裸鼠体内小细胞肺癌的生长和新生血管生成。
OBJECTIVE: To investigate the effect of 4-diphenylacetoxy-N-methylpiperidine methiodide, a muscarinic cholinergic receptor 3 (M3R) antagonist, DAMP) on the growth and angiogenesis of small cell lung cancer in nude mice. Methods: The expression of M3R mRNA and protein in human small cell lung cancer cell line SBC3 was detected by RT-PCR and Western blotting. A nude mouse xenograft model of human small cell lung cancer SBC3 cells was established and divided into 4 groups at random: 0.9% sodium chloride solution (control group) and 4-DAMP (0.5, 1 and 2 mg / kg) , Administered once daily for 15 days. The tumor volume was measured every 2 days. After the administration, the nude mice were killed, the tumor mass was measured, and the tumor inhibition rate was calculated. Real-time quantitative PCR and Western blotting were used to detect the expression of M3R and vascular endothelial growth factor (VEGF) mRNA and protein in each group. Immunohistochemistry was used to detect VEGF expression and microvessel density (MVD). Results: Human small cell lung cancer SBC3 cells expressed M3R mRNA and its protein. Successfully established a nude mouse xenograft model of human SBC3 cells and treated with different doses (0.5-2 mg / kg) of 4-DAMP, the volume of xenografts in nude mice was significantly decreased compared with the control group (P <0.05). The tumor inhibition rates of 4, 5, 1, and 2 mg / kg 4-DAMP groups were 15.82%, 33.50% and 55.06%, respectively. The mRNA and protein expressions of M3R and VEGF were significantly down-regulated in 0.5-2 mg / kg 4-DAMP group (all P <0.05), and MVD was also significantly decreased (all P <0.05). Conclusion: M3R antagonist 4-DAMP can inhibit the growth of small cell lung cancer and neovascularization in nude mice.