【摘要】 目的 分析对接蛋白3(DOK3)基因表达下调对原代结肠癌细胞增殖、侵袭/迁移能力的影响，并基于DOK3与G蛋白偶联受体84(GPR84)的相互作用探讨可能的作用机制。方法 培养原代结肠癌细胞HCT16,将其分为对照组、小干扰RNA(siRNA)-NC组、siRNA-DOK3组进行实验。分别将siRNA-NC、siRNA-DOK3转染至siRNA-NC组、siRNA-DOK3组，而仅将培养液加入对照组。培养24 h后通过实时荧光定量PCR评估转染效果，再使用CCK-8法、Transwell实验分别检测各组细胞的增殖情况、侵袭/迁移能力，采用蛋白免疫共沉淀实验检测HCT16细胞中DOK3与GPR84之间的相互作用关系。结果 与对照组比较，siRNA-DOK3组HCT16细胞中DOK3 mRNA表达量降低(P<0.05),而siRNA-NC组HCT16细胞中DOK3mRNA表达量差异无统计学意义(P>0.05)。培养48 h、72 h后，siRNA-DOK3组HCT16细胞的增殖活力较对照组增加(P<0.05)。与对照组相比，siRNA-DOK3组HCT16细胞的迁移和侵袭数量增加(P<0.05),而siRNA-NC组HCT16细胞的迁移和侵袭数量差异无统计学意义(P>0.05)。蛋白免疫共沉淀实验结果显示，在HCT16细胞中GPR84与DOK3相互结合。结论 DOK3基因可能作为抑癌基因，参与了结肠癌的发生与发展，且其可能通过与GPR84结合发挥抑癌作用。更多还原

【Abstract】 Objective To analyze the effect of down-regulated expression of docking protein 3(DOK3) gene on proliferation, invasion/migration abilities of primary colon cancerous cells, and to explore its possible mechanism based on interaction effects between DOK3 and G protein-coupled receptor 84(GPR84). Methods Primary colon cancerous cells HCT16 were cultured, and they were assigned to control group, small interfering RNA(siRNA)-NC group, or siRNA-DOK3 group, and then performing experiment. SiRNA-NC and siRNA-DOK3 were transfected into the siRNA-NC group and the siRNA-DOK3 group, respectively, whereas culture solution was only added into the control group. Transfection effect was evaluated by the real-time fluorescent quantitative PCR after 24 hours of culture, and then the CCK-8 method, Transwell experiment were used to detect proliferation, invasion/migration abilities of cells in various groups, respectively. Co-immunoprecipitation assay was used to detect interaction effects between DOK3 and GPR84 in HCT16 cells. Results Compared with the control group, the siRNA-DOK3 group exhibited a decreased mRNA expression of DOK3 in HCT16 cells(P<0.05), whereas no statistically significant difference in DOK3 mRNA expression in HCT16 cells between the control group and the siRNA-NC group(P>0.05). After 48-and 72-hour culture, proliferation activity of HCT16 cells in the siRNA-DOK3 group was increased as compared with the control group(P<0.05). Compare with the control group, the siRNA-DOK3 group yielded increased number of HCT16 cell migrations and invasions(P<0.05), whereas there was no statistically significant difference in the number of HCT16 cell migrations and invasions between the siRNA-NC group and the control group(P>0.05). The results of co-immunoprecipitation assay revealed that GPR84 interacted with DOK3 in HCT16 cells.Conclusion DOK3 gene may be regarded as a tumor suppressor gene, which participates in the occurrence and development of colon cancer, and it may exert a tumor suppressive effect by binding with GPR84.更多还原