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目的:研究硒化壳聚糖对慢性粒细胞白血病耐多柔比星细胞株(K562/DOX)增殖的影响,探讨其与磷脂酰肌醇-3激酶/丝氨酸-苏氨酸蛋白激酶(PI-3K/Akt)信号通路的关系。方法:将K562/DOX细胞分为阴性对照组、DOX组(50μg/mlDOX)和25、50、100、200、400μg/ml硒化壳聚糖(50μg/mlDOX+相应浓度的硒化壳聚糖)组,每组4个复孔,作用12、24、36h后,应用MTT法和克隆形成法检测硒化壳聚糖对细胞增殖的影响并计算逆转倍数,应用免疫印迹法检测DOX组和100、200μg/ml硒化壳聚糖组作用24h后细胞内磷酸化Ak(tp-Akt)蛋白表达。结果:与DOX组比较,50、100、200、400μg/ml硒化壳聚糖组对细胞具有明显抑制作用(P<0.05或P<0.01),且与浓度、时间呈正相关;25、50、100、200、400μg/ml硒化壳聚糖组细胞的逆转倍数分别为1.11、1.57、1.68、2.09、2.51。硒化壳聚糖明显下调了细胞内p-Akt蛋白表达(P<0.01)。结论:硒化壳聚糖可通过抑制PI-3K/Akt信号通路对K562/DOX细胞产生增殖抑制和多药耐药逆转作用。
OBJECTIVE: To study the effect of selenized chitosan on the proliferation of chronic myeloid leukemia cell line (K562 / DOX) and its relationship with phosphatidylinositol 3-kinase / 3K / Akt) signaling pathway. Methods: K562 / DOX cells were divided into negative control group, DOX group (50μg / ml DOX) and 25,50,100,200,400μg / ml selenium chitosan (50μg / ml DOX + corresponding concentration of chitosan) Group with 4 replicate wells for 12, 24 and 36 hours. MTT assay and colony formation assay were used to detect the effect of selenized chitosan on cell proliferation. The reversal multiples were calculated. DOX and 100, The expression of intracellular phosphorylated Akt (tp-Akt) protein was detected after treated with 200μg / ml selenium chitosan for 24 hours. Results: Compared with DOX group, the selenium chitosan groups at 50, 100, 200, 400μg / ml had significant inhibitory effect on the cells (P <0.05 or P <0.01), and were positively correlated with the concentration and time; The reversal multiples of cells in 100,200,400μg / ml selenized chitosan group were 1.11,1.57,1.68,2.09,2.51 respectively. Selenized chitosan downregulated the expression of p-Akt protein (P <0.01). Conclusion: Chitosan selenide can inhibit the proliferation of K562 / DOX cells and reverse the multidrug resistance by inhibiting the PI-3K / Akt signaling pathway.