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该文探讨了干扰肿瘤坏死因子受体相关因子6(tumor necrosis factor receptor-associated factor 6,TRAF6)表达对人白血病K562细胞增殖、凋亡的影响及其分子机制。将靶向TRAF6基因的sh RNA慢病毒载体感染K562细胞,利用荧光显微镜观察感染效率;Western blot方法检测TRAF6蛋白表达的改变;CCK-8法检测体外细胞增殖活性;流式细胞术分析细胞凋亡率;Western blot方法检测凋亡相关蛋白Bax、Bcl-2表达和AKT磷酸化水平的变化。结果显示,TRAF6-sh RNA慢病毒载体成功感染K562细胞,TRAF6蛋白表达水平明显下降。与空白对照组和TRAF6-NC组相比较,TRAF6-sh RNA组细胞增殖能力明显受抑(P<0.05),而细胞凋亡率增加(P<0.05);同时,促凋亡蛋白Bax表达升高、抗凋亡蛋白Bcl-2表达下降。此外,干扰TRAF6可下调K562细胞p-AKT(T308)和p-AKT(S473)活性,而总AKT水平未见明显变化。该研究表明,干扰TRAF6表达可抑制K562细胞增殖和诱导细胞凋亡,其机制可能与下调AKT活性有关,提示TRAF6可作为白血病治疗的一个潜在靶点。
This article explores the effects of interrupting the expression of tumor necrosis factor receptor-associated factor 6 (TRAF6) on the proliferation and apoptosis of human leukemia K562 cells and its molecular mechanism. Infection of K562 cells with sh RNA lentivirus vector targeting TRAF6 gene was performed to observe the infection efficiency by fluorescence microscopy. The expression of TRAF6 protein was detected by Western blot. The cell proliferation activity was detected by CCK-8 assay. The apoptosis of cells was analyzed by flow cytometry Western blot was used to detect the expression of Bax, Bcl-2 and AKT phosphorylation. The results showed that TRAF6-sh RNA lentiviral vector successfully infected K562 cells, TRAF6 protein expression decreased significantly. Compared with the blank control group and the TRAF6-NC group, the cell proliferation of TRAF6-sh RNA group was significantly inhibited (P <0.05), while the apoptosis rate was increased (P <0.05); at the same time, the expression of pro-apoptotic protein Bax High, anti-apoptotic protein Bcl-2 expression decreased. In addition, TRAF6 knockdown down-regulated the activity of p-AKT (T308) and p-AKT (S473) in K562 cells, but there was no significant change in total AKT levels. The study shows that disruption of TRAF6 expression can inhibit K562 cell proliferation and induce apoptosis, the mechanism may be related to the reduction of AKT activity, suggesting that TRAF6 can be used as a potential target for the treatment of leukemia.