辣椒素通过上调MTSS1表达抑制结肠癌细胞侵袭、迁移的实验研究

Capsaicin inhibits invasion and migration of colon cancer cells by upregulating MTSS1 expression

目的探究辣椒素通过上调肿瘤转移抑制因子1(MTSS1)抑制结肠癌细胞侵袭、迁移的机制。方法实验1:10、25、50、100μmol/L辣椒素组分别加入终浓度为10、25、50、100μmol/L辣椒素,对照组不添加辣椒素,恒温箱中培养24 h;实验2:对照组正常培养,100μmol/L辣椒素组添加终浓度为100μmol/L辣椒素,100μmol/L辣椒素+对照siRNA组、100μmol/L辣椒素+MTSS1 siRNA组在100μmol/L辣椒素组的基础上分别添加终浓度为50 nmol/L对照siRNA、MTSS1 siRNA,转染4 h更换为终浓度100μmol/L辣椒素培养液。CCK8检测细胞增殖情况;Transwell检测细胞侵袭情况;划痕实验检测细胞迁移情况;蛋白质免疫印迹(Western blot)检测细胞中辣椒素受体(TRPV1)、MTSS1、基质金属蛋白酶(MMP)-2、MMP-9蛋白水平。结果不同浓度辣椒素处理细胞发现,与对照组相比,25、50、100μmol/L辣椒素组细胞增殖抑制率升高,10、25、50、100μmol/L辣椒素组细胞中TRPV1、MTSS1蛋白水平升高,细胞侵袭数量、迁移百分比、细胞中MMP-2、MMP-9蛋白水平降低(P<0.05)。干扰MTSS1并添加100μmol/L辣椒素发现,与对照组相比,100μmol/L辣椒素组、100μmol/L辣椒素+对照siRNA组、100μmol/L辣椒素+MTSS1 siRNA组细胞侵袭数量、迁移百分比、细胞中MMP-2、MMP-9蛋白水平降低,细胞中MTSS1蛋白水平升高(P<0.05);分别与100μmol/L辣椒素组、100μmol/L辣椒素+对照siRNA组相比,100μmol/L辣椒素+MTSS1 siRNA组细胞侵袭数量、迁移百分比、细胞中MMP-2、MMP-9蛋白水平升高,细胞中MTSS1蛋白水平降低(P<0.05)。结论辣椒素通过上调MTSS1表达抑制结肠癌细胞侵袭、迁移,从而起到对结肠癌的保护作用。

Objective To explore the mechanism of capsaicin in inhibiting the invasion and migration of colon cancer cells by upregulating tumor metastasis suppressor 1(MTSS1). Methods Experiment 1: 10, 25, 50 and 100 μmol/L capsaicin groups were treated with capsaicin, whereas the control group was untreated and cultured at a constant temperature for 24 hours. Experiment 2: the control group was cultured normally, the 100 μmol/L capsaicin group was treated with 100 μmol/L capsaicin, 100 μmol/L capsaicin+control siRNA and 100 μmol/L capsaicin+MTSS1 siRNA groups were treated with 50 nmol/L control siRNA and MTSS1 siRNA, respectively, based on the 100 μmol/L capsaicin group. After transfection for 4 h, 100 μmol/L capsaicin-containing culture medium was added. Cell proliferation was assessed by CCK8 assays. Cell invasion was assessed by Transwell assays. Cell migration was assessed by a scratch test. The protein levels of transient receptor potential vanilloid 1(TRPV1), MTSS1, matrix metalloproteinase(MMP)-2, and MMP-9 were measured by Western blot. Results After treatment with capsaicin at various concentrations, compared with the control group, cell proliferation in 25, 50 and 100 μmol/L capsaicin groups was higher, the protein levels of TRPV1 and MTSS1 in 10, 25, 50 and 100 μmol/L capsaicin groups were higher, and cell invasion, the percentage of migrated cells and the protein levels of MMP-2 and MMP-9 were lower(P<0.05). After RNA interference of MTSS1 and addition of 100 μmol/L capsaicin, compared with the control group, cell invasion, the percentage of migrated cells, and the protein levels of MMP-2 and MMP-9 were lower in 100 μmol/L capsaicin, 100 μmol/L capsaicin+control siRNA, 100 μmol/L capsaicin+MTSS1 siRNA groups(P<0.05), and the protein level of MTSS1 was higher(P<0.05). Compared with 100 μmol/L capsaicin and 100 μmol/L capsaicin+control siRNA groups, cell invasion, the percentage of migrated cells, the protein levels of MMP-2 and MMP-9 were significantly higher in the 100 μmol/L capsaicin+MTSS1 siRNA group and the level of MTSS1 protein was lower(P<0.05). Conclusions Capsaicin inhibits the invasion and migration of colon cancer cells by upregulating the expression of MTSS1, thereby exerting a protective effect against colon cancer.