IL-22诱导卵巢癌细胞系SKOV3分泌CXCL13的机制研究

Mechanism of IL-22-induced CXCL13 secretion in ovarian cancer cell line SKOV3

目的探讨白细胞介素22(IL-22)诱导卵巢癌细胞系SKOV3细胞分泌CXCL13的分子机制。方法将体外培养的卵巢癌细胞系SKOV3细胞随机分为未刺激组、IL-22刺激组、对照组、DMSO组、NF-κB抑制剂组、JNK抑制剂组、p38/MAPK抑制剂组、MEK1/2抑制剂组以及STAT3抑制剂组。除未刺激组外,其余各组均用IL-22刺激细胞,流式细胞术检测SKOV3细胞表达的IL-22R水平,酶联免疫吸附试验(ELISA)检测CXCL13的水平,趋化实验检测T细胞的趋化效果。结果与未刺激组相比,IL-22刺激组SKOV3细胞分泌CXCL13的水平显著增加,差异具有极显著性统计学意义(P<0.01),并具有剂量和时间依赖性;在IL-22刺激的条件下,STAT3抑制剂组SKOV3细胞分泌CXCL13的水平显著低于DMSO组、NF-κB抑制剂组、JNK抑制剂组、MEK1/2抑制剂组和p38/MAPK抑制剂组,差异具有极显著性统计学意义(P<0.01);而NF-κB抑制剂组、JNK抑制剂组、MEK1/2抑制剂组和p38/MAPK抑制剂组与DMSO组比较,差异无统计学意义(P>0.05)。此外,IL-22刺激SKOV3细胞后的培养上清可诱导滤泡辅助性T细胞的趋化,而加入CXCL13中和抗体后趋化效果则显著下降(P<0.01)。结论IL-22经STAT3信号通路诱导卵巢癌细胞系SKOV3细胞分泌CXCL13,从而调控滤泡辅助性T细胞的趋化。

Objective To study the molecular mechanism of IL-22-induced CXCL13 secretion in ovarian cancer cell line SKOV3.Methods SKOV3 cells cultured in vitro were randomly divided into non-stimulated group,IL-22 stimulated group,control group,DMSO group,NF-κB inhibitor group,JNK inhibitor group,p38/MAPK inhibitor group,MEK1/2 inhibitor group and STAT3 inhibitor group.IL-22 was used to stimulate cells in all groups except the non-stimulated group,IL-22R expression in SKOV3 cells was detected by flow cytometry,the secretion of CXCL13 was detected by enzyme-linked immunosorbent assay.Chemotaxis assay was used to analyze the chemotactic movement of T cells. Results Compared with the non-stimulated group,the level of CXCL13 secreted by SKOV3 cells in IL-22 stimulated group was significantly increased,with statistically significant difference( P <0.01),which was in a dose and time dependent manner.However,the level of CXCL13 secretion induced by IL-22 in the STAT3 inhibitor group was significantly lower than that in DMSO group,NF-κB inhibitor group,JNK inhibitor group,MEK1/2 inhibitor group and p38/MAPK inhibitor group,with statistically significant differences ( P <0.01),but no statistical difference was observed between the DMSO group and NF-κB inhibitor group,JNK inhibitor group,MEK1/2 inhibitor group,p38/MAPK inhibitor group ( P >0.05).Additionally,follicular helper T cells were recruited by cell suspension obtained from SKOV3 cells stimulated by IL-22 ,and such chemotaxis was reversed while CXCL13 was neutralized. Conclusion IL-22 can regulate the chemotaxis of follicular helper T cells by inducing CXCL13 secretion through STAT3 signal pathway in ovarian cancer cell line SKOV3.