肝再生磷酸酶-3上调白细胞介素-6和白细胞介素-8促进结肠癌侵袭的机制研究

Phosphatase of regenerating liver-3 promoted the invasion of LoVo cells through interleukin-6 and interleukin-8

目的 观察细胞因子白细胞介素（IL）-6和IL-8在肝再生磷酸酶-3（PRL-3）诱导结肠癌细胞LoVo的侵袭和转移中的作用机制.方法 将实验分别空白对照组和实验转染组.酶联免疫吸附试验（ELISA）以及Western blot分别检测IL-6和IL-8蛋白的表达变化,IL-6和IL-8中和抗体（5 mg/L）处理细胞后,Transwell检测LoVo细胞增殖和侵袭的能力的变化.结果 将成功合成的PRL-3真核表达载体并转染入LoVo细胞中,ELISA结果提示结肠癌细胞稳定表达PRL-3后,PRL-3可明显促进结肠癌细胞分泌IL-6[（125±11） ng/L比（176±14） ng/L]和IL-8[（157±12） ng/L比（214 ±15） ng/L].Western blot结果提示PRL-3使IL-6的表达升高了3.2倍,IL-8的表达升高2.5倍（P＜0.05）.分别使用IL-6和IL-8中和抗体可以明显抑制PLR-3的促肿瘤侵袭效应[（78.0±3.5）个比（45.8±4.5）个,（78.0±3.5）个比（35.7±6.5）个].结论 PRL-3可以有效诱导结肠癌LoVo细胞分泌IL-6和IL-8,从而诱导结肠癌细胞迁移和侵袭.

Objective To investigate the effects cytokines of interleukin （IL）-6 and IL-8 in the invasion of LoVo cell line induced by phosphatase of regenerating liver-3 （PRL-3）.Methods LoVo cell lines were divided into two groups：the blank control group （LoVo-C）,the PRL-3 expression group （LoVo-P）.Enzyme linked immunosorbent assay （ELISA） and Western blot were used to detect the IL-6 and IL-8 protein levels.After treatment with IL-6 and IL-8 antibody （5 mg/L）,then the ability of invasion of LoVo cells was assessed by transwell invasion assay with the use of the IL-6 and IL-8 antibodies,respectively.Results In the LoVo-P group,the result of ELISA showed that the secretion of IL-6 [（125±11） ng/Lvs.（176±14） ng/L] andIL-8 [（157 ±12） ng/Lvs.（214 ±15） ng/L] were increased significantly and Western blot showed that PRL-3 enhanced the expression of IL-6 and IL-8 by 3.2-fold and 2.5-fold respectively （P ＜ 0.05）.When cells were treated with IL-6 and IL-8 antibody,the invasion ability of LoVo cells was inhibited significantly （78.0 ± 3.5 vs.45.8 ± 4.5,78.0 ± 3.5 vs.35.7 ± 6.5）.Conclusion PRL-3 could regulate the expression of IL-6 and IL-8 of LoVo cells line and could be a target to prevent the metastasis of colon cancer.