半枝莲抑制大肠癌干细胞自我更新及成瘤能力

目的探讨半枝莲对大肠癌干细胞自我更新和成瘤能力的影响及其可能的调控机制。方法采用侧群细胞分选法从SW480人结肠癌细胞中分离大肠癌干细胞。利用克隆球实验检测不同浓度半枝莲(0、0.25、0.5、0.75 mg/mL)干预24 h对大肠癌干细胞体外自我更新能力的影响;加入不同浓度半枝莲(0、0.5 mg/mL)干预大肠癌干细胞24 h后,采用BALB/c裸鼠皮下移植瘤实验检测半枝莲对大肠癌干细胞体内成瘤能力的影响;采用Western blot实验检测不同浓度半枝莲(0、0.25、0.5、0.75 mg/mL)干预24 h对大肠癌干细胞自我更新和成瘤能力标志物Nanog、SOX2、OCT4及Akt信号通路中p-Akt、Akt蛋白表达的影响。结果半枝莲可显著抑制大肠癌干细胞的体外自我更新能力,可见克隆球数量明显减少(P<0.05);半枝莲可明显抑制大肠癌干细胞的体内成瘤能力,可见半枝莲组裸鼠的移植瘤质量明显减轻(P<0.05);半枝莲可显著下调大肠癌干细胞Nanog、SOX2、OCT4及p-Akt的蛋白表达(P<0.05)。结论半枝莲可抑制大肠癌干细胞的自我更新和成瘤能力,其作用机制可能是通过抑制Akt信号通路的活化。

Scutellaria Barbata D. Don Inhibits the Self-renewal and Tumorigenic Ability of Colorectal Cancer Stem Cells

Objective: To explore the effects of Scutellaria barbata D. Don on the self-renewal and tumorigenesis ability of colorectal cancer stem cells(CR-CSCs) and its possible regulatory mechanism. Methods: The CR-CSCs were isolated from SW480 human colon cancer cells by side population cell sorting. The sphere formation assay was used to detect the effect of different concentrations(0, 0.25, 0.5, 0.75 mg/m L) of Scutellaria barbata D. Don on the self-renewal ability of CR-CSCs in vitro for 24 h. After stimulated CR-CSCs with Scutellaria barbata D. Don(0, 0.5 mg/m L) for 24 h, the BALB/c nude mice subcutaneous inoculation xenograft model was used to detect the effect of Scutellaria barbata D. Don on the tumorigenesis ability of CR-CSCs in vivo. Western Blot was used to detect the effect of Scutellaria barbata D. Don on the expression of the self-renewal and tumorigenesis ability markers Nanog,SOX2, OCT4 and the expression of p-Akt and Akt proteins in CR-CSCs after treatment with different concentrations(0, 0.25, 0.5,0.75 mg/m L) of Scutellaria barbata D. Don for 24 h. Results: Scutellaria barbata D. Don markedly inhibited the self-renewal ability of CR-CSCs in vitro, and the number of spheroids was significantly decreased(P<0.05). Scutellaria barbata D. Don profoundly inhibited the tumorigenesis ability of CR-CSCs in vivo, the weight of transplanted tumors of nude mice in the Scutellaria barbata D. Don group was significantly reduced(P<0.05). The protein expression of Nanog, SOX2, OCT4 and p-AKT in CR-CSCs were significantly down-regulated by Scutellaria barbata D. Don. Conclusion: Scutellaria barbata D. Don can inhibit the self-renewal and tumorigenesis ability of CR-CSCs, and its mechanism of action may be achieved by inhibiting the activation of Akt signaling pathway.