目的 研究分析结肠癌患者血管内皮生长因子(vascular endothelial growth factor,VEGF)信号通路的改变,以及五味子乙素对SW620细胞的影响,并分析其可能的作用机制。方法 蛋白印迹实验检测结肠癌患者正常肠道组织和癌变肠道组织中VEGFA、VEGF-R2、磷脂酰肌醇-3羟基激酶(phosphatidylinositol 3-kinase,PI3K)、Akt、p-Akt蛋白的表达,CCK-8法检测五味子乙素对SW620细胞增殖的影响,Transwell检测五味子乙素对SW620细胞迁移的影响;实时荧光定量PCR法检测SW620细胞内VEGFA、VEGF-R2、PI3K、Akt基因表达水平;免疫印迹法检测SW620细胞内VEGFA、VEGF-R2、PI3K、Akt、p-Akt蛋白表达水平。结果 与正常肠道组织相比,结肠癌患者癌变肠道组织内VEGF-R2、PI3K、Akt、p-Akt蛋白表达水平显著升高(P<0.05);与对照组比较,五味子乙素可以显著抑制SW620细胞的增殖和迁移(P<0.01),同时可以显著降低SW620细胞内VEGFA、VEGF-R2、PI3K、Akt基因表达水平和蛋白表达水平(P<0.01)。结论 结肠癌患者肠道内VEGF/PI3K/Akt信号通路被激活,五味子乙素可以抑制SW620细胞的活性和迁移,同时抑制VEGF/PI3K/Akt信号通路。
Abstract
OBJECTIVE To study the changes of VEGF signaling pathway in colon cancer patients and the effect of schisandrin B on SW620 cells and to analyze its possible mechanism. METHODS The protein expression of VEGFA, VEGF-R2, PI3K, Akt and p-Akt in human cancerous colon samples and adjacent normal samples were detected by Western blotting. The proliferation of SW620 cells was detected by CCK-8 method. The mRNA expression of VEGFA, VEGF-R2, PI3K and Akt in SW620 cells were detected by real-time PCR. The protein expression of VEGFA, VEGF-R2, PI3K, Akt and p-Akt in SW620 cells were detected by Western blotting. RESULTS The protein expression of VEGF-R2, PI3K, Akt and p-Akt in human cancerous colon samples was significantly higher than that in the adjacent normal samples(P<0.05). Compared with the control group, schisandrin B could significantly inhibit the proliferation and migration of SW620 cells, the mRNA expression of VEGFA, VEGF-R2, PI3K and Akt(P<0.01) and the protein expression of VEGFA, VEGF-R2, PI3K, Akt and p-Akt(P<0.05) in SW620 cells also were significantly decreased by schisandrin B. CONCLUSION The VEGF/PI3K/Akt signaling pathway is activated in colon cancer patients. Schisandrin B could inhibit the activity and migration of SW620 cells and inhibit the VEGF/PI3K/Akt signaling pathway.
关键词
结肠癌 /
SW620细胞 /
血管内皮生长因子 /
磷脂酰肌醇-3羟基激酶 /
Akt /
五味子乙素
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Key words
colon cancer /
SW620 cell /
vascular endothelial growth factor /
PI3K /
Akt /
schisandrin B
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中图分类号：
R965
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参考文献
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脚注
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基金
国家自然科学基金项目资助(81573685)
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