[关键词]
[摘要]
本文通过研究不同浓度6-姜烯酚诱导结肠癌细胞HT 29凋亡及与VEGFR2(血管内皮细胞受体2)不同表达之间的关系,以探讨其可能的抑制结直肠肿瘤相关机制。通过应用不同浓度6-姜烯酚(0、10、20和40 μM)分别对体外细胞实验培养的HT29细胞株进行诱导处理24 h, 相差倒置荧光显微镜观察细胞形态变化,CCK8(Cell Counting Kit-8)法测定药物诱导后细胞抑制率,Annexin V-FITC/PI流式细胞术检测不同浓度药物干预后细胞凋亡率及凋亡周期变化,最后用Western-blot检测分析VEGFR2蛋白不同表达变化。结果显示,与对照组相比,6-姜烯酚可呈浓度依赖性地诱导HT29凋亡,并使细胞周期G0/G1期细胞数由49.48%下降到24.39%和17.18%,而G2/M期细胞数由3.44%增加到34.78%和49.54%,进而将细胞周期阻滞在G2/M期,抑制细胞增殖,而且还不同程度地抑制了VEGFR2表达(p<0.05)。结果表明,6-姜烯酚主要是通过诱导HT29凋亡进而起到抑制肿瘤细胞增殖的作用,这可能与不同程度抑制VEGFR2有关。
[Key word]
[Abstract]
In order to unravel the mechanism of the inhibition of colorectal cancer by 6-shogoal, the apoptosis of colon cancer cells HT29 induced by different concentrations of 6-shogoal, as well as its relationship with different expression of VEGFR2 (vascular endothelial cell receptor2) was investigated. The HT29 cells were treated with different concentrations of 6-shogoal (0, 10, 20 and 40 μM) cultured in vitro. The cell morphology changes were observed by the contrast inverted fluorescence microscope, and the cell inhibition rate was determined by CCK8 method. Annexin V-FITC/ PI flow cytometry was used to detect the changes of apoptosis rate and apoptosis cycle. Finally, the expression of VEGFR2 protein was detected by Western-blot. The results showed that as compared with the control group, 6-shogoal could induce HT29 apoptosis in a concentration dependent manner. The number of cell cycle G0/G1 cells decreased from 49.48% to 24.39% and 17.18%, while the number of G2/M cells increased from 3.44% to 34.78% and 49.54%.Thus,the cell cycle was blocked in G2/M stage to inhibit the cell proliferation. Besides, it also inhibited the expression of VEGFR2 to different extents (p<0.05).Therefore, it can be concluded that 6-shogoal can induce HT29 apoptosis and inhibit the proliferation of tumor cells, which may be related to different degrees of inhibition of VEGFR2.
[中图分类号]
[基金项目]
国家自然科学基金项目(81460369);宁夏医科大学优势学科群项目(2001060304)