具核梭杆菌促进食管癌细胞增殖的研究

Fusobacterium nucleatum promotes the proliferation of esophageal cancer cells

目的研究具核梭杆菌（Fusobacteriumnucleatum，n）对食管癌细胞粘附入侵和增殖的作用，探讨其作用机制。方法本实验应用具核梭杆菌标准株ATCC25586与食管癌Eca-109细胞共培养，建立具核梭杆菌与食管癌细胞体外的肿瘤微环境模型。采用细胞免疫组化染色法检测细菌对细胞的粘附入侵能力[3-（4，5-二甲基噻唑-2）-2，5-二苯基四氮唑溴盐]MTF及流式细胞仪检测Eca-109细胞的增殖，酶联免疫吸附试验方法（ELISA）检测观察正常对照组、细菌与细胞共培养模型组的细胞培养液中IL-6的浓度，Westernblot检测STAT3和p-STAT3的蛋白表达。结果具核梭杆菌可粘附和入侵到Eca-109细胞内。与对照组比较，加入具核梭杆菌的实验组作用后第1—4天Eca-109细胞增殖加快，48h内其细胞培养液内IL-6浓度也是对照组的4倍。Westernblot结果显示，与对照组相比，具核梭杆菌处理组p-STAT3蛋白表达增加，STAT3蛋白表达没有明显改变。中和IL-6受体蛋白后，具核梭杆菌处理组p-STAT3、STAT3蛋白表达量均不变。结论具核梭杆菌通过刺激IL．6的产生促进食管癌细胞的增殖。

Objective To study the Fusobacterium nucleatum（F, nucleatum） function on the role of esophageal cancer cell, to explore its mechanism. Methods F. nucleatum standard strains ATCC 25586 were co-cultured with esophageal cancer Eca-109 cells to establish the tumor microenvironment model of bacteria with host cells in vitro, bacterial invasion and adhesion ability were detected by immunohistochemical staining; Eca-109 cell proliferation was assay by flow cytometry and M＇IT; IL-6 concentration was detected by enzyme-linkedimmunosorbent assay （ELISA） in the cell supematant of normal control group, bac- teria and cell co-culture model group respectively; the protein expression of STAT3 and p-STAT3 were detected by Western blot. Results F. nucleatum can adhere and invade into Eca-109 cells. Compared with the control group, the experimental group with F. nucleatum had accelerated Eca-109 cell proliferation in 1-4 days. The concentration of IL-6 in cell culture medium also had elevated 4 times in 48 h. Western blot results showed that F. nucleatum-treated group increased the expression of p-STAT3 protein, but STAT3 protein expression did not change significantly compared with the control group. After neutralizing IL-6 receptor protein, the expression of p-STAT3, STAT3 protein were unchanged in F. nucleatum-treated group. Conclusion F. nucleatum can promote the proliferation of esophageal cancer cells by stimulating IL-6 production.