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目的研究Skp2基因表达下调对HepG2细胞生物学特性及p27和SP1蛋白表达的影响。方法构建Skp2基因干扰慢病毒质粒,包装后感染HepG2细胞,48 h后,流式细胞仪检测细胞周期和凋亡情况;侵袭小室试验检测细胞侵袭能力;Western blot检测Skp2基因下调后p27和SP1蛋白的表达情况。结果与正常HepG2细胞组相比,Skp2干扰慢病毒组细胞Skp2蛋白的表达明显下调,p27蛋白表达大幅增加,细胞生长速度明显减慢,细胞凋亡率增加近两倍,G0/G1期细胞增多;SP1蛋白表达减少,浸润、迁移的细胞数减少约50%(P<0.05)。结论 Skp2蛋白表达下调抑制了HepG2细胞的增殖,促进HepG2细胞的凋亡,造成细胞侵袭能力减弱。Skp2蛋白表达下调导致p27蛋白表达上调、SP1蛋白表达下调,在上述生物学特性变化中可能发挥了重要作用,为深入探讨肝癌的病理机制奠定了基础。
Objective To investigate the effect of down-regulation of Skp2 gene on the biological characteristics of HepG2 cells and the expression of p27 and SP1 proteins. Methods Skp2 gene was constructed to interfere with the lentiviral plasmid and infected HepG2 cells after packaging. After 48 h, the cell cycle and apoptosis were detected by flow cytometry. Invasion chamber assay was used to detect cell invasion. Western blot was used to detect the expression of p27 and SP1 protein The expression of the situation. Results Compared with normal HepG2 cells, the expression of Skp2 protein in Skp2 interfered lentivirus group was significantly down-regulated, the expression of p27 protein was significantly increased, the cell growth rate was slowed down, the apoptosis rate nearly doubled and the number of G0 / G1 phase cells increased ; The number of SP1 protein decreased, the number of infiltrating and migrating cells decreased about 50% (P <0.05). Conclusion The down-regulation of Skp2 protein can inhibit the proliferation of HepG2 cells and promote the apoptosis of HepG2 cells, resulting in the decrease of cell invasion ability. The down-regulation of Skp2 expression leads to the up-regulation of p27 protein and down-regulation of SP1 protein, which may play an important role in the above biological changes and lay a foundation for further study on the pathological mechanism of hepatocellular carcinoma.