受体相互作用蛋白3对神经胶质瘤U251细胞增殖影响.docVIP

受体相互作用蛋白3对神经胶质瘤U251细胞增殖影响 　　[摘要] 目的 探讨受体相互作用蛋白3（RIP3）在神经胶质瘤细胞中的表达及其对U251细胞增殖的影响。 方法 采用实时定量PCR（real-time RT-PCR）法分别检测神经胶质瘤A172、U251、U373和U87细胞中RIP3的表达水平，随后应用U251稳定过表达RIP3（U251-RIP3）细胞模型，分别采用MTT法和克隆形成实验检测细胞增殖能力，流式细胞仪评价细胞周期变化。结果 RIP3 mRNA在4株胶质瘤细胞（A172、U251、U373、U87）中均有表达，并在U251细胞中表达最低。real-time RT-PCR结果表明U251-RIP3细胞中稳定过表达RIP3，与阴性对照（空载）（NC）相比，RIP3过表达能够抑制U251细胞增殖能力；能够降低U251细胞克隆形成能力，培养11 d后细胞集落数目（67±2）?^NC（75±3）减少（P 0.05）；能够延缓细胞周期进程，U251-RIP3细胞的S期比例较NC组（16.0±0.4）%升至（18.2±1.0）%（P 0.05），同时G0/G1期比例较NC组（55.7±0.5）%降至（57.4±0.4）%（P 0.01）。 结论 RIP3过表达后可抑制神经胶质瘤U251细胞增殖，有望成为靶向控制胶质瘤细胞增殖的潜在治疗靶点。 　　[关键词] 受体相互作用蛋白3；胶质瘤；细胞增殖；可控性坏死 　　[中图分类号] R739.41 [文献标识码] A [文章编号] 1673-7210（2017）04（a）-0004-04 　　[Abstract] Objective To investigate the expressive changes of RIP3 in all sorts of glioma cell lines and the effects on the cell proliferation in the U251 cells. Methods The expression levels of RIP3 mRNA in the four glioma cell lines （A172， U251， U373 and U87） were firstly selected by real-time RT-PCR. Then， U251 cells were engineered to overexpress RIP3 for further study. Lastly， the MTT， colony formation， and flow cytometry assays were used to detect the effects of RIP3 on the cell proliferation， clone formation， and cell cycle of U251 cells， respectively. Results The expression levels of RIP3 mRNA was detected in all four glioma cell lines， and the lowest level of it was discovered in U251 cell line. Then， real-time RT-PCR assay showed that RIP3 was successfully overexpressed in U251-RIP3 cells compared with negative control （NC） cells with empty vector. The overexpressed RIP3 altered the cell viability in U251 cells， Additionally， the overexpressed RIP3 also reduced the colony formation of U251 cells： after 11 d of cell culture， the colony number of U251-RIP3 cells （67±2） was significantly decreased compared with that of NC cells （75±3） （P 0.05）. Correspondingly， the proportion of U251-RIP3 cells in S phase arrest was higher than that in NC cells [from （16.0±0.4）% to （18.2±1.0）% ] （