中文摘要：

目的观察蛋白酶体激活因子REGgamma（REGγ）基因对乳腺癌MDA-MB-231细胞生长的影响。方法构建真核表达重组体PcDNA3．1-REGγ并将其以脂质体转染法导人细胞，以600mg／L浓度的G418连续筛选转染细胞6周，获得稳定高表达该基因的细胞株。分别以噻唑蓝（MTT）、流式细胞仪（FCM）、软琼脂集落形成试验检测其对细胞生长的影响。免疫细胞化学检测增殖细胞核抗原（PCNA）的表达。结果转染REGγ的细胞有外源REGγ基因的整合及表达，经Westernblot检测其表达较未转染组和仅转染空载体组明显增加。MTT法检测发现转染REGγ细胞生长加速；FCM检测提示转染REGγ组、未转染组和仅转染空载体组在S＋G2＋M增殖期的细胞比例分别为55．91％、44．09％、43．69％；软琼脂集落形成试验显示其平均集落形成率分别为10．23％、3．67％、4．06％。转染REGγ基因后PCNA表达明显增强。结论转染外源野生型REG7基因对乳腺癌MDA-MB-231细胞具有加速其生长、促进其增殖的作用。

英文摘要：

Objective To study the effect of REGgamma gene on growth of human breast carcinoma MDA-MB-231 cells. Methods The eukaryotic expression recombinant PcDNA3. 1-REGgamma gene was constructed and tansfected into human breast carcinoma MDA-MB-231 cells by lipofectamine 2000. The cells transed with empty vector served as controls. The stably transfected cells were selected after exposure to 600 mg/L C,418 for 6 weeks. The effects of REGgamma gene on growth of human breast carcinoma MDA-MB-231 cells were examined by MTF, flow cytometry （ FCM ） and soft agar conoly formation test. Immunocytochemistry was performed to detect the expression of proliferative cell nuclear antigen （PCNA） in MDA-MB-231 ceils. Results The expression of REGgamma gene was highly increased after transfection. Cell growth was accelerated and the proportion of cells in proliferative phase in tansfected with REG- gamma gene group, untransfected group and transfeeted with empty vector group was 55.91% ,4,＊. 09%0 and 43.69%0 respectively. The cell colony forming efficiency was 10.23% ,3.67% and 4.06% in tansfected with REGgamma gene group, untransfected group and transfeeted with empty vector group respectively. The expression of PCNA was markedly enhanced after transfection. Conclusion Cell growth and proliferation were accelerated after MDA-MB-231 was transfected with REGgamma gene.