目的?探究肿瘤相关基因羧肽酶A4(CPA4)对肝癌细胞增殖、迁移和侵袭的影响及其相关分子机制。方法?慢病毒载体构建MHCC97L-CPA4过表达细胞系和MHCC97H-shCPA4敲减细胞系，分别以MHCC97L-vector和MHCC97H-shNC细胞系作为对照。采用实时荧光定量PCR(qRT-PCR)和western blot检测各组细胞中CPA4?mRNA和蛋白质的表达；CCK8试验和克隆形成试验检测各组细胞的体外增殖能力；Transwell试验检测细胞的体外迁移、侵袭能力；western blot检测各组细胞STAT3/p-STAT3/c-myc的表达差异。结果?与vector组相比，CPA4组MHCC97L细胞的增殖、迁移、侵袭能力均升高(P<0.05)；与shNC组相比，shCPA4组MHCC97H细胞的增殖、迁移、侵袭能力均降低(P<0.05)。western blot结果显示，与vector组相比，CPA4组MHCC97L细胞p-STAT3、c-myc蛋白的表达水平升高(P<0.05)；与shNC组相比，shCPA4组MHCC97H细胞p-STAT3、c-myc蛋白的表达水平降低(P<0.05)。结论?CPA4可能通过激活STAT3信号通路促进肝癌细胞的增殖、迁移和侵袭。
Objective?To explore the effects of carboxypeptidase A4 (CPA4) on proliferation, migration and invasion of hepatoma cells and its related molecular mechanism.?Methods?MHCC97L-CPA4 overexpression cell line and MHCC97H-shCPA4 knockdown cell line were constructed by lentiviral vector. MHCC97L-vector and MHCC97H-shNC cell lines were used as control respectively. Real-time fluorescent quantitative PCR (qRT-PCR) and western blot were used to detect the expressions of?CPA4?mRNA and protein in each group of cells. CCK8 kitand clone formation test were used to detect the proliferation ability in each cell group in vitro. The transwell assay was used to detect cell migration and invasion ability in vitro. western blot was used to detect the expression differences of STAT3/p-STAT3/c-myc in each group.?Results?Compared with the vector group, the proliferation, migration and invasion ability of MHCC97L cells in CPA4 group were increased (P<0.05). Compared with the shNC group, the proliferation, migration and invasion ability of MHCC97H cells in shCPA4 group were reduced (P<0.05). western blot results showed that, the expression levels of p-STAT3 and c-myc protein in MHCC97L cells in CPA4 group increased (P<0.05) compared with the vector group. p-STAT3 and c-myc in MHCC97H cells in shCPA4 group were decreased compared with the shNC group (P<0.05).?Conclusion?CPA4 may promote the proliferation, migration and invasion of liver cancer cells by activating the STAT3 signaling pathway.