槲皮素调控JAK2/STAT3对直肠癌细胞生物活性的影响

Mechanism of quercetin on regulating the biological activity of the JAK2/STAT3 signaling pathway on rectal cancer cells

目的:了解影响直肠癌细胞的作用机制,探讨槲皮素对JAK2/STAT3信号通路的调控作用,为直肠癌临床治疗提供依据。方法:人直肠癌Colo320细胞随机分为4组,包括A组(无药物干预Colo320细胞)、B组(Colo320细胞+25μmol/L的槲皮素)、C组(Colo320细胞+50μmol/L的槲皮素)及D组(Colo320细胞+100μmol/L的槲皮素),经不同浓度槲皮素干预后分别采用MTT法检测Colo320存活率,流式细胞仪检测Colo320凋亡率,Transwell实验检测侵袭数目,免疫印迹及RT-PCR检测JAK2/STAT3表达。结果:与A组相比,B组、C组及D组Colo320细胞存活率均下降(P<0.05),经不同浓度槲皮素干预的B、C及D组Colo320细胞存活率逐渐降低,D组72 h存活率最低,为(36.20±2.65)%,任意两组比较差异有统计学意义(P<0.05)。与A组相比,经不同浓度槲皮素处理的B组、C组及D组Colo320细胞凋亡率升高(P<0.05),D组Colo320细胞凋亡率最高,为(24.92±2.51)%,与B组、C组相比差异有统计学意义(P<0.05)。与A组相比,经不同浓度槲皮素处理的B组、C组及D组Colo320侵袭数量逐渐降低(P<0.05),D组Colo320细胞侵袭数量最低,为(44±10)个,与B组和C组比较差异有统计学意义(P<0.05)。与A组相比,B组Colo320细胞中JAK2/p-JAK2、STAT3/-pSTAT3蛋白及JAK2、STAT3 m RNA表达降低,组间比较差异有统计学意义(P<0.05);与B、C组相比,D组JAK2、STAT3蛋白、m RNA及磷酸化表达均降低(P<0.05)。结论:槲皮素能够降低直肠癌Colo320存活率及侵袭能力,增加凋亡,存在浓度依赖性,可能与抑制JAK2/STAT3磷酸化表达相关。

Objective: To understand the mechanism of effect on colorectal cancer cells and explore the regulation of quercetin on JAK2/STAT3 signaling pathway, so as to provide evidence for clinical treatment of rectal cancer.Methods: Colo320 cells from human rectal cancer were randomly divided into four groups, Colo320 cells without drug intervention, Group A(Colo320 cells +25μmol/L quercetin), Group C(Colo320 cells +50μmol/L quercetin) and Group D(Colo320 cells +100μmol/L quercetin), Colo320 survival rate was measured by MTT assay, apoptosis rate of Colo320was measured by flow cytometry, invasion number was measured by Transwell assay, and JAK2/STAT3 expression was measured by western blotting and RT-PCR.Results: Compared with group A, the survival rate of Colo320 cells in groups B, C, and D decreased(P<0.05). The survival rate of Colo320 cells in groups B, C, and D decreased gradually after treatment with different concentrations of quercetin. The survival rate of Colo320 cells in group D was the lowest at 72 h(36.20±2.65) %. There was statistically significant difference between any two groups(P<0.05).Compared with group A, the apoptosis rate of Colo320 cells in groups B, C and D was increased(P<0.05), and the apoptosis rate of Colo320 cells in group D was the highest(24.92±2.51) %,which was statistically significan(P<0.05) compared with groups B and C.Compared with group A,the number of Colo320 invasions in groups B, C, and D decreased gradually(P<0.05), and the number of Colo320 invasions in group D was the lowest(44±10), with statistically significant differences compared with groups B and C(P<0.05). Compared with group A, JAK2/P-JAK2 and STAT3/-pSTAT3 expression and JAK2 and STAT3 m RNA in Colo320 cells of group B were decreased, with statistically significant differences(P<0.05). Compared with groups B and C, the protein, m RNA and phosphorylation of JAK2 and STAT3 in group D were decreased(P<0.05). Conclusions: Quercetin can reduce the survival rate and invasion ability of Colo320 colorectal cancer and increase apoptosis in a dose-dependent manner, which may be associated with inhibition of JAK2/STAT3 phosphorylation expression.