双歧杆菌通过IL-6介导EMT对HCT116结肠癌细胞侵袭转移影响

Effect of Bifidobacterium on invasion and metastasis of HCT116 colon cancer cells through IL-6 mediated EMT

目的探讨双歧杆菌通过白细胞介素-6(IL-6)介导上皮间质转化(EMT)对HCT116结肠癌细胞侵袭转移的影响。方法体外培养HCT116结肠癌细胞,将其分为对照组和双歧杆菌组,对照组常规培养,双歧杆菌组加入80μg/ml的双歧杆菌培养。48 h后采用Western blot检测两组IL-6、上皮钙黏素和神经钙黏素蛋白表达水平,免疫荧光技术检测IL-6、上皮钙黏素和神经钙黏素的表达及定位。采用细胞划痕试验、Transwell小室实验检测两组细胞转移和侵袭能力。结果Western blot检测结果显示,双歧杆菌组上皮钙黏素蛋白表达水平高于对照组,IL-6、神经钙黏素蛋白表达水平低于对照组,差异有统计学意义(P<0.05)。免疫荧光检测结果显示,IL-6、上皮钙黏素和神经钙黏素蛋白定位于细胞膜上,与对照组比较,双歧杆菌组上皮钙黏素荧光明显,IL-6、神经钙黏素荧光微弱。双歧杆菌组细胞划痕愈合率低于对照组,差异有统计学意义(P<0.05)。Transwell小室实验结晶紫染色及定量分析结果显示,双歧杆菌组侵袭细胞数少于对照组,差异有统计学意义(P<0.05)。结论双歧杆菌可抑制炎症因子IL-6介导的EMT过程,减弱HCT116结肠癌细胞侵袭和转移能力。

Objective To investigate the effect of Bifidobacterium on invasion and metastasis of HCT116 colon cancer cells through interleukin-6(IL-6)mediated epithelial-mesenchymal transition(EMT).Methods HCT116 colon cancer cells were cultured in vitro and divided into control group and Bifidobacterium group.The control group was cultured conventionarily,and the Bifidobacterium group was cultured with 80μg/ml Bifidobacterium.After 48 hours,the expression levels of IL-6,E-cadherin,and N-cadherin in both groups were detected by Western blot,and the expression and localization of IL-6,E-cadherin,and N-cadherin were detected by immunofluorescence technique.Cell scratch test and Transwell chamber test were used to detect the cell metastasis and invasion ability of the two groups.Results The results of Western blot showed that the expression level of E-cadherin in the Bifidobacterium group was higher than that in the control group,while the expression level of IL-6 and N-cadherin was lower than that in the control group,the difference was statistically significant(P<0.05).The results of immunofluorescence showed that IL-6,E-cadherin,and N-cadherin proteins were localized on cell membranes.Compared with the control group,the fluorescence of E-cadherin in Bifidobacterium group was obvious,while the fluorescence of IL-6 and N-cadherin was weak.The healing rate of cell scratches in Bifidobacterium group was lower than that in control group,and the difference was statistically significant(P<0.05).The results of crystal violet staining and quantitative analysis in Transwell chamber test showed that the number of invasive cells in Bifidobacterium group was less than that in control group,and the difference was statistically significant(P<0.05).Conclusion Bifidobacterium can inhibit the EMT process mediated by inflammatory factor IL-6,and weaken the invasion and metastasis ability of HCT116 colon cancer cells.