毒蕈碱型胆碱受体拮抗剂抑制小细胞肺癌生长及血管生成的体内研究

发布时间：2018-10-20 17:28

【摘要】：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。
[Abstract]:Aim: to investigate the effects of 4-diphenylethoxy-N-methyl-piperidine iodide (4-diphenylacetoxy-Nmethylpiperidine methiodide,4-DAMP), an antagonist of muscarinic choline receptor 3 (muscarinic cholinergic receptor 3 (M3R), 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 blot. Nude mice with human small cell lung cancer (SBC3) were randomly divided into 4 groups: 0.9% sodium chloride solution (control group) and 4-DAMP4 at different doses (0.5 mg/kg and 2 mg/kg) respectively, once a day for 15 days. The tumor volume was measured once every 2 days, the nude mice were killed after administration, the tumor mass was measured and the tumor inhibition rate was calculated. The expression of M3R and vascular endothelial growth factor (vascular endothelial growth factor,VEGF) mRNA and protein in tumor tissues were detected by real-time fluorescence quantitative PCR and Western blot respectively. The expression of VEGF and microvessel density (microvessel density,MVD) were detected by immunohistochemistry. Results: human small cell lung cancer (SBC3) cells expressed M 3R mRNA and its protein. The transplanted tumor model of human SBC3 cells in nude mice was successfully established and treated with different doses of 4-DAMP (0.5 mg/kg). It was found that the volume of transplanted tumor in nude mice was significantly smaller than that of the control group (P < 0. 05), and the tumor quality of exfoliation was also significantly decreased (P = 0. 01). The tumor inhibition rates of 0.5 mg/kg 4-DAMP and 2 mg/kg 4-DAMP groups were 15.82% and 55.06% respectively. The expression of mRNA and protein in VEGF was significantly down-regulated (P < 0. 05), MVD, P < 0. 05). Conclusion: M3R antagonist 4-DAMP can inhibit the growth and angiogenesis of small cell lung cancer in nude mice.
【作者单位】：宁夏医科大学研究生院;宁夏医科大学总医院呼吸与危重症医学科;
【基金】：国家自然科学基金资助项目(编号:81260356)~~
【分类号】：R734.2

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