Lu Xingrong, Yi Jilin
Department of General Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
J Huazhong Univ Sci Technolog Med Sci. 2005;25(3):289-91. doi: 10.1007/BF02828145.
In order to investigate the effect of small ubiquitin-like modifier-1 (SUMO-1) on the p53-induced HepG2 cell apoptosis, HepG2 cells were transfected by recombinant plasmids as pwtp53, pMDM2 and pSUMO-1 respectively. Western blot was employed to detect the protein expression of the transfected recombinant plasmids and the rate of apoptosis was measured by flow cytometry. The results showed that in cells transfected with pwtp53 and pwtp53 + pSUMO-1, the apoptosis rate was (16.79+/-1.62) % and (18.15+/-1.36) % respectively, while transfected with pwtp53+ pMDM2, the rate was decreased to (5.17+/-1.23) %. The apoptosis rate was (14.06+/-1.84) % in the cells transfected with pwtp53+pMDM2+pSUMO-1, significantly higher than that in the cells Transfected with pwtp53+pMDM2 (P<0.01). The apoptosis rates in the cells were all less than 2 % and had no significant difference among the groups. It was suggested that in the HepG2 cells, SUMO-1 can increase the apoptosis induced by wild-type p53 through binding to p53 protein, posttranslational modification and inhibiting the p53 degradation by MDM2.
为了研究小泛素样修饰物1(SUMO-1)对p53诱导的HepG2细胞凋亡的影响,分别用重组质粒pwtp53、pMDM2和pSUMO-1转染HepG2细胞。采用蛋白质免疫印迹法检测转染重组质粒后的蛋白表达情况,并用流式细胞术检测细胞凋亡率。结果显示,转染pwtp53和pwtp53 + pSUMO-1的细胞凋亡率分别为(16.79±1.62)%和(18.15±1.36)%,而转染pwtp53 + pMDM2的细胞凋亡率降至(5.17±1.23)%。转染pwtp53 + pMDM2 + pSUMO-1的细胞凋亡率为(14.06±1.84)%,显著高于转染pwtp53 + pMDM2的细胞(P<0.01)。各细胞组凋亡率均小于2%,组间无显著差异。提示在HepG2细胞中,SUMO-1可通过与p53蛋白结合、进行翻译后修饰并抑制MDM2对p53的降解,从而增加野生型p53诱导的细胞凋亡。