藤梨根对RKO结肠癌细胞失巢凋亡的作用

目的:观察藤梨根对人结肠癌RKO细胞悬浮生长及失巢凋亡作用.方法:200～800 mg·L-1藤梨根作用于RKO细胞,CCK-8(CCK-8)法检测藤梨根对RKO细胞悬浮生长作用,双嵌入剂乙锭均二聚物(EthD-1)染色检测失巢凋亡,比色法检测半胱天冬酶-3(caspase-3)活性,2-7-二氯氢化荧光素乙二脂染色法结合荧光酶标仪检测细胞内活性氧的产生.结果:200 ～800 mg·L-1藤梨根可显著抑制RKO悬浮生长,呈剂量依赖性(P＜0.01).200 ～800 mg·L-1藤梨根作用后悬浮生长RKO细胞吸收EthD-1散发红色荧光,部分细胞可见核碎裂,呈现凋亡形态改变;藤梨根组EthD-1荧光吸光度与对照组比较差异显著(P＜0.01).终质量浓度200 ～800 mg·L-1藤梨根可显著增强RKO细胞caspase-3活性以及细胞内活性氧的产生(P＜0.01).结论:藤梨根可抑制RKO细胞悬浮生长,促使RKO细胞发生失巢凋亡,可能与活化caspase-3以及升高细胞内活性氧水平相关.

Root of Actinidia chinensis Planch Induces Anoikis in Colon Carcinoma RKO Cells

Objective: To observe the effects of Tengligen (root of Actinidia chinensis Planch) on cell growth in suspension and anoikis in human colon carcinoma RKO cells. Method: RKO cells were treated with Tengligen (200-800 mg·L^-1) or same volume of RPMI1640.Cell growth in suspension was detected with CCK-8 assay. Anoikis was visualized by EthD-1 staining. Caspase-3 activities were detected by colorimetric assay. Intracellular reactive oxygen species (ROS) production was observed by 2', 7'-dichlorofluorescin diacetate staining. Result: 200-800 mg·L^-1 of Tengligen, significantly inhibited RKO cell growth in suspension in a dose dependent manner (P<0.01). After 200-800 mg·L^-1 of treatment, EthD-1 was absorbed by suspension-cultured RKO cells to yield a red-fluorescent nuclear staining, and nuclear fragmentation was observed in partial cells. The EthD-1 fluorescence in 200-800 mg·L^-1 of Tengligen treated RKO cells were compared with controls (P<0.01). 200-800 mg·L^-1 of Tengligen also activated caspase-3 and upregulated intracellular ROS generation (P<0.01). Conclusion: Tengligen could inhibit RKO cell growth in suspension, induce anoikis in RKO cells, and may associate with caspase-3 activation and ROS production.