Wnt1诱导分泌蛋白-1通过NF-κB通路促进食管鳞癌细胞的增殖和侵袭转移

目的: 探讨Wnt1诱导分泌蛋白 1(Wnt1 induced secreted protein 1， WISP 1)在食管鳞癌细胞侵袭转移的作用机制。方法: 体外培养人食管鳞癌细胞株Eca109、TE 8和正常食管上皮细胞Het 1a，荧光定量PCR和蛋白质印迹检测WISP 1表达。将食管鳞癌细胞株Eca109和TE 8分为空白对照组、阴性对照组和WISP 1 siRNA组，利用CCK8法检测各组细胞增殖改变；流式细胞术分析不同处理组细胞凋亡率；Transwell实验检测细胞侵袭能力变化；蛋白质印迹法检测细胞中VEGF A，VEGF C，MMP2，MMP9以及NF κB通路相关蛋白的表达量；酶联免疫吸附法测定培养上清液中VEGF C和MMP9分泌量。结果： 荧光定量PCR和蛋白质印迹结果显示，食管鳞癌细胞中WISP 1表达量高于正常食管上皮细胞(P<0.01)；CCK8和流式细胞术结果显示，下调WISP 1后，食管鳞癌细胞株增殖明显抑制（P<0.05），凋亡率无明显影响；蛋白质印迹结果显示，下调WISP 1对VEGF A和MMP2 表达无明显影响，而VEGF C和MMP9表达明显得到抑制；酶联免疫吸附结果显示，VEGF C和MMP9分泌量随WISP 1下调也出现明显下降（P<0.05）；下调WISP 1后，食管鳞癌细胞株侵袭能力明显降低（P<0.01）；下调WISP 1表达显著抑制NF κB磷酸化并促进IκBα的磷酸化，抑制NF κB信号活化水平。 结论： WISP 1可通过NF κB通路，增加MMP9，VEGF C的表达和分泌，促进食管鳞癌细胞增殖及侵袭转移的能力。

WISP-1 promotes the invasion and metastasis of esophageal squamous cell carcinoma by NF-κB signaling pathway

Objective: To investigate the role and mechanism of Wnt1 induced secreted protein 1(WISP 1) in invasion and metastasis of esophageal squamous cell carcinoma. Methods: Human esophageal squamous cell carcinoma cells(ESCC), Eca109 and TE 8, and normal human esophageal epithelial cells, Het 1a, were cultured in vitro. RT PCR and Western blotting were used to detect the expression of WISP 1 between ESCCs and Het 1a cells. Eca109 and TE 8 cells were divided into blank control, negative control and WISP 1 siRNA groups. The cell proliferation was detected by CCK8 method. Flow cytometry was used to analyze the apoptosis rate of each group. Transwell assay was used to detect the ability of invasion. The expression of VEGF A, VEGF C, MMP2 and MMP9 in each group was detected by Western blotting. The secretion of VEGF C and MMP9 were detected by ELISA. The expression changes of NF κB pathway were determined by Western blotting. Results: WISP 1 were highly expressed in ESCCs compared with Het 1a cells(P<0.01). CCK8 and flow cytometry results showed that the proliferation of ESCCs were significantly inhibited by down regulation of WISP 1(P<0.05), while had no significant effect on the apoptosis. The results of Western blotting and ELISA showed that, even though there were no significant change of VEGF A and MMP2, the expression or secretion of VEGF C and MMP9 were down regulated after siRNA WISP 1 treatment(P<0.05). Down regulation of WISP 1 could significantly reduce the phosphorylation of NF κB p65 and induce the phosphorylated IκBα(p IκBα), but had no effect on the total expression of NF κB and IκBα. Conclusion: WISP 1 promotes the invasion and metastasis of ESCC via NF κB pathway.