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目的:探讨再生基因Ⅳ(regenerating gene Ⅳ,RegⅣ)及其碳水化合物识别域(carbohydrate-recognition domain,CRD)对裸鼠结直肠癌移植瘤血管生成的影响。方法:将已转染了重组质粒pcDNA3.1-RegⅣ、pcDNA3.1-Reg Ⅳ△ CRD、空载体质粒的人结肠癌LoVo细胞(分别命名为LoVo/RegⅣ、LoVo/Reg Ⅳ△ CRD、LoVo/negative)和自然生长状态的LoVo细胞分别接种于裸鼠皮下。RT-PCR法检测移植瘤组织中RegⅣ及Reg Ⅳ△ CRDmRNA的表达,免疫组织化学法检测移植瘤组织中血管内皮生长因子(vascularendothelial growth factor,VEGF)和CD34的表达,计数移植瘤组织中微血管密度(microvessel density,MVD)。结果:LoVo/RegⅣ移植瘤组织中VEGF的表达水平高于其他3组,差异有统计学意义(P<0.05);LoVo/Reg Ⅳ△ CRD、LoVo和LoVo/negative组间VEGF的表达水平差异无统计学意义(P>0.05);LoVo/RegⅣ移植瘤组织中MVD值明显高于其他3组(P<0.05),LoVo/Reg Ⅳ△ CRD、LoVo和LoVo/negative移植瘤组织间MVD值差异无统计学意义(P>0.05)。结论:RegⅣ基因参与调控结直肠癌的血管生成,其作用与CRD结构域密切相关。
Objective: To investigate the effect of regenerating gene Ⅳ (Reg Ⅳ) and its carbohydrate-recognition domain (CRD) on the angiogenesis of colorectal cancer xenografts in nude mice. Methods: Human colon cancer LoVo cells (named as LoVo / RegⅣ, LoVo / Reg Ⅳ △ CRD, LoVo / Reg Ⅳ △ CRD) were transfected with recombinant plasmid pcDNA3.1-RegⅣ, pcDNA3.1- negative) and LoVo cells in their natural growth were inoculated subcutaneously in nude mice respectively. The expression of Reg Ⅳ and Reg Ⅳ △ CRD mRNA in the xenograft tumor was detected by RT-PCR. The expression of VEGF and CD34 in the xenografted tumor was detected by immunohistochemistry. The microvessel density (microvessel density, MVD). Results: The expression of VEGF in LoVo / RegIV xenografts was significantly higher than that in the other three groups (P <0.05). There was no significant difference in VEGF expression between LoVo / Reg Ⅳ △ CRD, LoVo and LoVo / negative groups (P> 0.05). The MVD in LoVo / RegIV xenografts was significantly higher than that in the other three groups (P <0.05). There was no significant difference in MVD among LoVo / Reg Ⅳ △ CRD, LoVo and LoVo / negative xenografts Statistical significance (P> 0.05). Conclusion: Reg Ⅳ gene is involved in the regulation of angiogenesis in colorectal cancer. Its role is closely related to the CRD domain.