薏苡仁油诱导细胞周期阻滞及细胞凋亡对结肠癌细胞化疗敏感性的影响

Coix seed oil enhances the chemosensitivity of colon cancer cells by inducing cell cycle arrest and apoptosis

目的探究薏苡仁油对结肠癌细胞化疗敏感性的影响。方法体外培养人结肠癌HT29细胞。使用不同浓度的薏苡仁油(1、2、4、8 mg/ml)与30μg/ml 5-氟尿嘧啶(5-FU)共孵育HT29细胞24 h模拟化疗,采用MTT法和流式细胞仪检测各组细胞增殖抑制率、细胞凋亡率、细胞周期占比,Western blot测定各组细胞中cleaved caspase-3蛋白表达。结果1、2、4、8 mg/ml薏苡仁油+5-FU组细胞增殖抑制率显著高于5-FU组和薏苡仁油组(P<0.05),其中2、4、8 mg/ml薏苡仁油+5-FU组显著高于1 mg/ml薏苡仁油+5-FU组(P<0.05);5-FU组、薏苡仁油组及1、2、4、8 mg/ml薏苡仁油+5-FU组细胞凋亡率显著高于空白对照组(P<0.05),1、2、4、8 mg/ml薏苡仁油+5-FU组细胞凋亡率显著高于5-FU组和薏苡仁油组(P<0.05),2、4、8 mg/ml薏苡仁油+5-FU组细胞凋亡率显著高于1 mg/ml薏苡仁油+5-FU组(P<0.05);各组cleaved caspase-3表达结果与流式细胞术检测的凋亡率的结果基本一致;5-FU组、薏苡仁油组及1、2、4、8 mg/ml薏苡仁油+5-FU组G1/M期细胞的占比显著高于空白对照组(P<0.05),S期细胞的占比显著低于空白对照组(P<0.05);1、2、4、8 mg/ml薏苡仁油+5-FU组G1/M期细胞的占比显著高于5-FU组和薏苡仁油组(P<0.05),S期细胞的占比显著低于5-FU组和薏苡仁油组(P<0.05);2、4、8 mg/ml薏苡仁油+5-FU组G1/M期细胞的占比显著高于1 mg/ml薏苡仁油+5-FU组(P<0.05),S期细胞的占比显著低于1 mg/ml薏苡仁油+5-FU组(P<0.05)。结论薏苡仁油可能通过诱导细胞周期阻滞及细胞凋亡增强结肠癌细胞化疗敏感性。

Objective To investigate the effect of coix seed oil on chemosensitivity of colon cancer cells.Methods HT29 cell line was cultured in vitro.Different concentrations of coix seed oil(1,2,4,8 mg/ml)and 30μg/ml 5-fluorouracil(5-FU)were incubated with HT29 cells for 24 hours to simulate chemotherapy.The cell proliferation inhibition rate,apoptosis rate and cell cycle ratio were measured by methyl thiazolyl tetrazolium(MTT)method and flow cytometry,and the protein expression of cleaved caspase-3 was measured by Western blot.Results The inhibition rate of cell proliferation in 1,2,4,8 mg/ml coix oil+5-FU group was significantly higher than that in 5-FU group and coix oil group,and the inhibition rate in 2,4,8 mg/ml coix oil+5-FU group was significantly higher than that in 1 mg/ml coix oil+5-FU group(P<0.05).The apoptosis rate in 5-FU group,coix oil group and 1,2,4,8 mg/ml coix oil+5-FU group was higher than that in the blank control group(P<0.05).The apoptosis rate in 1,2,4,8 mg/ml coix oil+5-FU group was significantly higher than that in 5-FU group and coix oil group(P<0.05).The apoptosis rate of 2,4,8 mg/ml coix oil+5-FU group was significantly higher than that of 1 mg/ml coix oil+5-FU group(P<0.05).The expression of cleaved caspase-3 in each group was basically in line with the apoptosis rate of flow cytometry.The percentage of G1/M phase cells in 5-FU group,coix oil group and 1,2,4,8 mg/ml coix oil+5-FU group was significantly higher than that in the blank control,and the percentage of S phase cells was lower comparing with blank control(P<0.05).Besides,the percentage of G1/M phase cells in 1,2,4,8 mg/ml coix oil+5-FU group was significantly higher than that in 5-FU group and coix oil group,and the percentage of S phase cells was significantly lower than that in 5-FU group and coix oil group(P<0.05).The percentage of G1/M phase cells in 2,4,8 mg/ml coix oil+5-FU group was significantly higher than that in 1 mg/ml coix oil+5-FU group,and the percentage of S phase cells was significantly lower than that in 1 mg/ml coix oil+5-FU group(P<0.05).Conclusions Coix seed oil may enhance the chemosensitivity of colon cancer cells by inducing cell cycle arrest and apoptosis.