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目的探讨肝再生磷酸酶-3(PRL-3)对人肺癌细胞侵袭能力的影响及相关机制。方法采用RT-PCR和Western blot检测高低转移能力不同的4种肺癌细胞及HBE细胞PRL-3 mRNA及蛋白的表达。免疫荧光检测A549细胞、HBE细胞中PRL-3的定位。PRL-3特异性小干扰RNA(siRNA)转染高转移肺癌细胞株A549后,采用RT-PCR和Western blot检测肺癌细胞PRL-3和mDia1 mRNA及蛋白表达,MTT、迁移试验和Transwell检测肺癌细胞系增殖及迁移侵袭能力,G-LISA方法检测RhoA活性。结果肺癌细胞系(A549,CI-H661)PRL-3 mRNA及蛋白表达高于HBE细胞。A549细胞系中PRL-3定位于细胞质,而HBE细胞系中PRL-3则定位于细胞核周围。PRL-3 siRNA能够显著下调A549细胞PRL-3 mRNA和蛋白表达水平,引起RhoA活性降低及其下游作用物mDia1的表达降低,A549细胞的迁移侵袭能力降低(P<0.01)。结论 PRL-3的表达与肺癌细胞系的迁移侵袭能力有关,PRL-3可能通过调控RhoA活性及mDia1表达促进肺癌细胞迁移侵袭。
Objective To investigate the effect of liver regeneration phosphatase-3 (PRL-3) on invasion of human lung cancer cells and its related mechanisms. Methods The expression of PRL-3 mRNA and protein in four kinds of lung cancer cells and HBE cells with different metastatic potentials were detected by RT-PCR and Western blot. Immunofluorescence was used to detect the location of PRL-3 in A549 cells and HBE cells. The expression of PRL-3 and mDia1 mRNA and protein in lung cancer cells were detected by RT-PCR and Western blot after PRL-3 specific small interfering RNA (siRNA) transfected A549 cells. MTT, migration assay and Transwell assay were used to detect lung cancer cells Department of proliferation and migration of invasive ability, G-LISA method to detect RhoA activity. Results PRL-3 mRNA and protein expression in lung cancer cell lines (A549, CI-H661) were higher than those in HBE cells. PRL-3 localized in the cytoplasm in the A549 cell line, whereas PRL-3 in the HBE cell line localized around the nucleus. PRL-3 siRNA significantly down-regulated the mRNA and protein expression of PRL-3 in A549 cells, decreased the activity of RhoA and decreased the expression of mDia1, and decreased the migration and invasion of A549 cells (P <0.01). Conclusion The expression of PRL-3 is related to the migration and invasion of lung cancer cell lines. PRL-3 may promote the migration and invasion of lung cancer cells through the regulation of RhoA activity and mDia1 expression.