【作者】 罗庚求；

【导师】 李景和；

【作者基本信息】 中南大学 ， 病理学与病理生理学， 2003， 硕士

【摘要】 目的：研究DPC4基因转染对结肠癌血管生成的影响，探讨DPC4对结肠癌的生长抑制作用及其可能作用机制。 方法：利用脂质体介导转染技术建立表达Smad4蛋白的DPC4^+-SW620（PcDNA-DPC4转染的SW620^*细胞）细胞模型；利用免疫细胞化学S-P法和Western blot检测细胞中Smad4的表达；利用ELISA法检测细胞上清中VEGF蛋白的表达；利用RT-PCR检测细胞内VEGFmRNA的表达；利用皮下注射法建立裸鼠结肠癌移植瘤模型；利用免疫组织化学S-P法检测裸鼠肿瘤组织中VEGF的表达及微血管密度。 结果：1、建立了表达Smad4蛋白的DPC4⁺-SW620细胞系； 2、DPC4⁺-SW620细胞，其Smad4蛋白表达于细胞浆及细胞核，以胞浆为主；DPC4⁺-SW620细胞其Smad4蛋白表达水平明显高于SW620细胞及PcDNA-SW620细胞（PcDNA3.1转染的SW620细胞）； 3、DPC4⁺-SW620细胞其VEGF蛋白、VEGFmRNA水平明显低于SW620细胞及PcDNA-SW620细胞（P＜0.05）； 4、成功地建立了裸鼠结肠癌移植瘤模型；DPC4⁺-SW620组裸鼠肿瘤生长速度慢于SW620细胞组及PcDNA-SW620细胞组（P＜0.05）；DPC4⁺-SW620组裸鼠肿瘤重量低于SW620细胞组及PcDNA-SW620细胞组（P＜0.05）； 硕士学位论文 摘要 5、DPC4”－SW620组裸鼠肿瘤组织VEGF蛋白、月瘤组织微血管 密度均低 于 SW620细胞组及 PCDNA－SW620细胞组（HO．05）； 结论1、DPC4能够抑制裸鼠结肠癌移植瘤的生长； 2、通过抑制肿瘤的血管生成可能是DPC4发挥对结肠癌的抑制 作用的机制之一。更多还原

【Abstract】 Objective:To study the mechanism of transfected DPC4 gene on angiogenesis and growth suppression in human colon carcinoma .Methods:SW620 cells, a human colorectal carcinoma cell line, were transfected with PcDNA3.1-DPC4 plasmid using lipofectamine transfecting technique. Expressing Smad4 in DPC4+-SW620 cells was observed by Immunohistochemistry( SP method) and Western blot. The expression of VEGF protein in the cell supernatant was detected by ELISA, VEGF mRNA by RT-PCR. The model of nude mice inoculated with DPC4+-SW620 cells was established by injecting into flank subcutaneously.The expression of VEGF protein and the microvessel density of tumor tissue in nude mice were detected by Immunohistochemistry( SP method).Results:1. DPC4+-SW620 cells expressing Smad4 were harvested.2. Smad4 Protein showed stronger expression in SW620 cells transfected with PcDNA-DPC4 plasmid than that in not-transfected SW620 cells and SW620 cells transfected with blank plasmid, and the positive signal was localized in cytoplasm and nucleus, mainly in cytoplasm.3. There were lower expressions of VEGF protein and mRNA in DPC4’ - SW620 cells than in SW620 cells and PcDNA-SW620 cells(P<0.05).4. The model of nude mice inoculated with colon carcinoma cells was established successfully. The tumors of nude mice inoculated with DPC4-SW620 cells growed more slowly than that inoculated with SW620 cells and PcDNA-SW620 cells. The volume and weight of tumors in nude mice inoculated with DPC4 ’ ~sw620 cells were smaller than that with sw620 cells and PcDNA-sw620 cells(P<0.05).5. The expressions of VEGF and the microvessel density in DPC4’ -SW620 cells were lower than that in SW620 cells and PcDNA-SW620 cells(p<0.05).Conclusions:1 .The DPC4 gene can suppress the tumors of nude mice inoculated with DPC4 + -SW620 cells.2.The inhibition effect of DPC4 on colon carcinoma may be partly mediated by suppressing angiogenesis of tumor.更多还原