肝再生磷酸酶3对SGC7901细胞迁移侵袭及RhoC表达的影响

Influence of phosphatase of regenerating liver 3 on cell migration,invasion and expression of RhoC in SGC7901 cells

目的:通过观察肝再生磷酸酶3(phosphatase of regenerating liver 3,PRL-3)对胃癌SGC7901细胞迁移侵袭及RhoC表达的影响,探讨胃癌SGC7901细胞迁移侵袭的机制中PRL-3和RhoC是否位于同一信号通路.方法:体外培养人胃癌SGC7901细胞,经不同浓度PRL-3抗体(1∶600,1∶400,1∶200)作用后,采用细胞划痕实验观察不同时间段(0、12、24、48 h)SGC7901细胞的迁移距离;通过Transwell迁移侵袭实验检测不同浓度PRL-3抗体(1∶600,1∶400,1∶200)作用48 h后SGC7901细胞迁移侵袭的穿膜细胞数,并采用Real-time PCR和ELISA检测SGC7901细胞中RhoC mRNA及蛋白表达量的变化,比较不同浓度PRL-3抗体处理后SGC7901细胞迁移侵袭的细胞数和RhoC mRNA及蛋白的表达量.结果:与对照组比较,随着PRL-3抗体浓度的升高和作用时间的延长,SGC7901细胞的迁移距离逐渐减小,在48 h,经不同浓度PRL-3抗体(1∶600,1∶400,1∶200)处理SGC7901细胞后,其迁移和侵袭的细胞数,RhoCmRNA相对表达量和蛋白表达水平与对照组相比均明显降低(迁移SGC7901细胞数:365.0±5.0,165.3±5.0,90.3±5.5 vs 512.3±4.9;侵袭SGC7901细胞数:321.3±6.1,179.0±6.1,75.7±4.0 vs 545.3±5.0;RhoC mRNA的相对表达量:0.910±0.022,0.742±0.018,0.539±0.015 vs 1.000±0.000;RhoC蛋白表达量:1130.77 g/mL±15.32 g/mL,981.52 g/mL±14.44 g/mL,893.03 g/mL±11.10 g/mL vs 1212.42 g/mL±18.37g/mL)(均P<0.01),不同PRL-3抗体浓度组两两比较均有统计学意义(均P<0.01).结论:PRL-3在体外能显著促进SGC7901细胞的迁移侵袭,并能增强RhoC基因和蛋白的表达,提示在SGC7901细胞迁移侵袭的机制中PRL-3和RhoC可能位于同一信号通路.

AIM：To explore whether phosphatase of regenerating liver 3（PRL-3） and RhoC belong to the same signal pathway in the mechanism of SGC7901 cell migration and invasion by observing the influence of PRL-3 on cell migration,invasion and the expression of RhoC in SGC7901 cells.METHODS：Human gastric cancer SGC7901 cells were cultured in vitro and treated with different concentrations of PRL-3Ab（1：600,1：400 and 1：200）,then the migration distance of SGC7901 cells was assessed at different time points（0,12,24,48 h） by wound healing assay,and the migration and invasion of SGC7901 cells were examined by Transwell assay at 48 h.The expression of RhoC mRNA and protein was detected by real-time PCR and ELISA.RESULTS：Compared with the control group,the migration distance of SGC7901 cells at 12,24 and 48 h decreased with the increase in PRL-3Ab concentration.At 48 h,the migration and invasion of SGC7901 cells and the expression of RhoC mRNA and protein in SGC7901 cells treated with different concentrations of PRL-3Ab（1：600,1：400 and 1：200） were signifi cantly lower than those in the control group（migration：365.0 ± 5.0,165.3 ± 5.0,90.3 ± 5.5vs 512.3 ± 4.9;invasion：321.3 ± 6.1,179.0 ± 6.1,75.7 ± 4.0 vs 545.3 ± 5.0;expression of RhoC mRNA：0.910 ± 0.022,0.742 ± 0.018,0.539 ±0.015 vs 1.000 ± 0.000;expression of RhoC protein：1130.77 g/mL ± 15.32 g/mL,981.52g/mL ± 14.44 g/mL,893.03 g/mL ± 11.10 g/mL vs 1212.42 g/mL ± 18.37 g/mL;P 0.01 for all）.CONCLUSION：PRL-3 can promote SGC7901 cell migration and invasion and raise the expression of RhoC,which suggests that PRL-3and RhoC may belong to the same signal pathway in the mechanism of SGC7901 cell migration and invasion.