| 摘要: |
| 目的 探讨川楝素（TSD）调节Ras/Raf/丝裂原激活蛋白激酶激酶（MEK）/细胞外信号调节激酶（ERK）信号通路对结肠癌细胞恶性生物学行为的影响。方法 将SW620细胞分为对照组（NC组）、低剂量TSD组（TSD-L组，1μmol/L）、中剂量TSD组（TSD-M组，2 μmol/L）、高剂量TSD组（TSD-H组，4 μmol/L）、ML-099（Ras/Raf/MEK/ERK通路激活剂）组（5 μmol/L）、TSD-H+ML-099组（4 μmol/L+5 μmol/L）。CCK-8法、克隆形成实验检测SW620细胞增殖；流式细胞术检测细胞凋亡；Transwell检测细胞迁移和侵袭；western blot检测SW620细胞中细胞周期蛋白D1(cyclinD1)、Bcl-2蛋白相关X蛋白(Bax)、基质金属蛋白酶（MMP）-9、Ras、Raf、p-MEK1/2、p-ERK1/2蛋白表达。裸鼠体内移植瘤实验检测TSD对裸鼠肿瘤质量与体积的影响。结果 与NC组比较，TSD-L组、TSD-M组、TSD-H组SW620细胞OD450值、克隆形成率、迁移与侵袭细胞数目、cyclinD1、MMP-9、Ras、Raf、p-MEK1/2、p-ERK1/2蛋白表达、裸鼠肿瘤质量与体积降低，细胞凋亡率、Bax蛋白表达升高，ML-099组对应指标变化趋势与上述相反（P＜0.05）；与TSD-H组比较，TSD-H+ML-099组SW620细胞OD450值、克隆形成率、迁移与侵袭细胞数目、cyclinD1、MMP-9、Ras、Raf、p-MEK1/2、p-ERK1/2蛋白表达、裸鼠肿瘤质量与体积升高，细胞凋亡率、Bax蛋白表达降低（P＜0.05）。结论 TSD可能通过抑制Ras/Raf/MEK/ERK信号通路抑制SW620细胞增殖、迁移、侵袭及裸鼠体内肿瘤生长，促进细胞凋亡。 |
| 关键词: 川楝素 结肠癌 Ras/Raf/丝裂原激活蛋白激酶激酶（MEK）/细胞外信号调节激酶（ERK）信号通路 侵袭 增殖 |
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| Impact of toosendanin on the malignant biological behavior of colon cancer cells by regulating Ras/Raf/MEK/ERK signaling pathway |
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Han Meng-yuan, Chen Jia-li, Xi Yan, Wang Dong-ming
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Shanghai University of Traditional Chinese Medicine Affiliated Longhua Hospital
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| Abstract: |
| Objective To investigate the impact of toosendanin (TSD) on the malignant biological behavior of colon cancer cells by regulating Ras/Raf/mitogen-activated protein kinase (MEK)/extracellular signal-regulated kinase (ERK) signal pathway. Methods SW620 cells were grouped into control group (NC group), low-dose TSD group (TSD-L group, 1 μmol/L), medium-dose TSD group (TSD-M group, 2 μmol/L), high-dose TSD group (TSD-H group, 4 μmol/L), ML-099 (Ras/Raf/MEK/ERK pathway activator) group (5 μmol/L), and TSD-H+ML-099 group (4 μmol/L+5 μmol/L). The proliferation of SW620 cells was detected by CCK-8 and clonogenic assay; cell apoptosis was detected by flow cytometry; Transwell was applied to detect the cell migration and invasion; Western blot was applied to detect the expression of cyclin D1, Bcl-2 protein associated X protein (Bax), matrix metalloproteinase-9, Ras, Raf, p-MEK1/2, p-ERK1/2 in SW620 cells. The effect of TSD on the mass and volume of tumor in nude mice was detected by tumor transplantation in vivo.Results Compared with NC group, the OD450 value of SW620 cells, the clonogenic rate, the numbers of migrating and invading cells, the expression of cyclinD1, MMP-9, Ras, Raf, p-MEK1/2, p-ERK1/2 proteins, the tumor mass and volume of nude mice in TSD-L group, TSD-M group and TSD-H group decreased, the apoptosis rate and the expression of Bax protein increased, the change trend of corresponding indicators in ML-099 group was contrary to the above (P<0.05); compared with TSD-H group, the OD450 value of SW620 cells, the clonogenic rate, the number of migrating and invading cells, the expression of cyclinD1, MMP-9, Ras, Raf, p-MEK1/2, p-ERK1/2 proteins, the tumor mass and volume of nude mice in TSD-H+ML-099 group increased, the apoptosis rate and the expression of Bax protein decreased (P<0.05).Conclusion TSD may inhibit the proliferation, migration, invasion and tumor growth of SW620 cells and promote cell apoptosis by inhibiting Ras/Raf/MEK/ERK signal pathway. |
| Key words: toosendanin colon cancer Ras/Raf/mitogen-activated protein kinase kinase (MEK)/extracellular signal-regulated kinase (ERK) signal pathway invasion proliferation |