牙龈卟啉单胞菌通过CXCL2/CXCR2轴促进口腔鳞癌进展的动物模型研究

An animal model study on promotion of oral squamous cell carcinoma by Prophyromonas gingivalis via axis of CXCL2/CXCR2

目的探讨牙龈卟啉单胞菌(Porphyromonas gingivalis,P.gingivalis)通过口腔鳞癌细胞中趋化因子配体2/趋化因子受体2(CXCL2/CXCR2)轴促进口腔鳞癌进展的机制。方法体外培养SCC7鼠源性鳞状细胞癌细胞株,P.gingivalis建立共培养模型,用C57BL/6小鼠建立荷瘤模型,用不同浓度CXCL2/CXCR2轴抑制剂SCH527123进行干预,实验分为对照组、高剂量组、低剂量组和实验组,于第10、20天两个时间点处死小鼠,测量各组瘤体体积,免疫组化检测Gαi、P.gingivalis、CXCL2、CD66b+、N-钙黏蛋白(N-Cad)、E-钙黏蛋白(E-Cad)在各组中的表达情况,用Elisa法检测CXCL2在各组中的含量。结果实验组瘤体体积较对照组明显增大(P<0.05),用CXCL2/CXCR2轴抑制剂干预时,与实验组相比,低剂量组瘤体体积缩小(P<0.05),高剂量组瘤体体积缩小更明显(P<0.01)。Gαi在实验组中的表达强于对照组、高剂量组和低剂量组(P<0.05),低剂量组与高剂量组相比差异有统计学意义,表明CXCL2/CXCR2信号轴抑制剂SCH527123成功抑制该信号轴。P.gingivalis、CXCL2、CD66b+、N-Cad在实验组中阳性或强阳性表达;CXCL2和CD66b+在对照组中阴性表达;P.gingivalis和CXCL2在高剂量组和低剂量组中弱阳性表达或阳性表达;E-Cad在对照组、高剂量组、低剂量组、实验组中分别是强阳性、阳性、弱阳性、阴性表达。实验组中CXCL2的含量最高,与对照组、高剂量组、低剂量组相比差异均有统计学意义(P<0.001)。结论P.gingivalis通过上调CXCL2的表达来促进肿瘤进展,用CXCL2/CXCR2轴抑制剂干预后,肿瘤的进展被抑制。

Objective To explore the mechanism of Porphyromonas gingivalis(P.gingivalis)promoting the progression of oral squamous cell carcinoma(OSCC)through the chemokine ligand 2/chemokine receptor 2(CXCL2/CXCR2)axis in oral squamous cell carcinoma cells.Methods The SCC7 murine squamous cell carcinoma cell line was cultured in vitro.The cell and bacteria co-culture model was established with using SCC7 and P.gingivalis.The tumor-bearing model was established in C57BL/6 mice.Different concentrations of CXCL2/CXCR2 axis inhibitor SCH527123 were used for intervention.The animals were divided into four groups as following:control group,high dose group,low dose group and experimental group respectively.The mice were euthanized at two time points on the 10th and 20th days,and the tumor volume of each group was measured.The expression of Gαi,P.gingivalis,CXCL2,CD66b+,N-Cadherin and E-Cadherin in each groups was tested by immunohistochemistry,and CXCL2 levels were tested by Elisa.Results Tumor volume was significantly larger in experimental group than that in control group(P<0.05).When using CXCL2/CXCR2 axis inhibitors,compared with experimental group,the tumor volume in low dose group was decreased(P<0.05),and the tumor volume in high dose group was decreased more significantly(P<0.01).The expression of Gαi in experimental group was stronger than that in control group,high dose group and low dose group(P<0.05).The difference between low dose group and high dose group was statistically significant,indicating that CXCL2/CXCR2 signal axis inhibitor SCH527123 successfully inhibited the signal axis.P.gingivalis,CXCL2,CD66b+,N-Cadherin were expressed positively or strongly in the experimental group;CXCL2 and CD66b+were negatively expressed in the control group;P.gingivalis and CXCL2 were weakly or positively expressed in high-dose and low-dose groups.E-Cadherin was expressed strong positively,moderate positively,weak positively,negatively in control group,high dose group,low dose group and experimental group respectively.CXCL2 levels were highest in experimental group,which was statistically significant compared with control group high dose group and low dose group(P<0.001).Conclusion P.gingivalis promotes tumor progression by up-regulating the expression of CXCL2.When CXCL2/CXCR2 axis inhibitor is used for intervention,tumor progression is inhibited.