PLCE1基因对胃癌细胞侵袭转移的影响

Influence of PLCE1 gene on invasion and metastasis of gastric cancer cells

目的探讨干扰磷脂酶Cε1(phospholipase C epsilon 1,PLCE1)基因对胃癌细胞侵袭转移的影响及相关机制。方法采用免疫组化SP法检测63例胃癌组织中PLCE1的表达,分析PLCE1表达与胃癌临床病理特征的关系;将胃癌SGC7901细胞株随机分为SiPLCE1组和对照组,分别转染PLCE1基因干扰慢病毒和阴性对照空载体慢病毒,采用细胞划痕实验和Transwell实验观察细胞侵袭迁移能力;采用qPCR和Western blot法检测细胞PLCE1 mRNA和蛋白表达情况;采用Western blot法检测细胞上皮-间质转化(epithelial-mesenchymal transition,EMT)相关蛋白(E-cadherin、N-cadherin、vimentin)以及PKC/ERK信号通路蛋白的表达情况。结果PLCE1高表达组胃癌患者的淋巴结转移率为37.93%、远处转移率为20.69%、Ⅲ~Ⅳ期患者占41.38%,均高于PLCE1低表达组(P<0.05)。细胞实验结果显示,Sh-PLCE1组24 h细胞迁移距离为(36.81±2.77)μm,细胞迁移数为(291.15±20.47)个,细胞侵袭数为(142.94±19.05)个,均明显低于对照组和ShRNA-NC组(P<0.05),PLCE1 mRNA和蛋白表达、N-cadherin、vimentin蛋白表达水平均低于对照组和ShRNA-NC组(P<0.05),E-cadherin蛋白表达水平高于对照组和ShRNA-NC组(P<0.05);Sh-PLCE1组PKCα、p-ERK/ERK蛋白表达水平均低于对照组和ShRNA-NC组(P<0.05)。结论PLCE1高表达可能与胃癌淋巴结转移、远处转移相关,抑制PLCE1基因可降低胃癌SGC7901细胞侵袭、迁移能力,其作用机制可能是通过抑制PKC/ERK信号通路进而抑制EMT。

Purpose To explore the influence of phospholipase C epsilon 1(PLCE1)gene interference on invasion and metastasis of gastric cancer cells and its related mechanism.Methods The expression of PLCE1 in tissues of 63 cases of gastric cancer was detected by immunohistochemistry,and the relationship between PLCE1 expression and clinicopathological characteristics of gastric cancer was analyzed.The gastric cancer SGC7901 cell lines were randomly divided into SiPLCE1 group and control group.The PLCE1 gene interference lentivirus and the negative control unloaded lentivirus were respectively transfected.Cell scratch assay and Transwell test were used to observe the cell invasion and migration.qPCR and Western blot were applied to detect the mRNA and protein expressions of PLCE1 gene.The expressions of epithelial-mesenchymal transition(EMT)-related proteins(E-cadherin,N-cadherin,vimentin)and PKC/ERK signaling pathway proteins were detected by Western blot.Results Statistical analysis showed that the lymph node metastasis rate,distant metastasis rate and proportion of patients with stageⅢ-Ⅳwere 37.93%,20.69%and 41.38%in high-expression PLCE1 group,which were higher than those in low-expression PLCE1 group(P<0.05).Cell experiments showed that the 24h cell migration distance,the number of cell migration and the number of cell invasion of(36.81±2.77)μm,(291.15±20.47)cells and(142.94±19.05)cells in Sh-PLCE1 group were significantly lower than those in control group and ShRNA-NC group(P<0.05),and the mRNA expression and protein expression of PLCE1 gene and protein expression levels of N-cadherin and vimentin were lower than those in control group and ShRNA-NC group(P<0.05)while the protein expression level of E-cadherin was higher than that in control group and ShRNA-NC group(P<0.05).The protein expression levels of PKCαand p-ERK/ERK in Sh-PLCE1 group were lower compared with those in control group and ShRNA-NC group(P<0.05).Conclusion The high expression of PLCE1 may be related to lymph node metastasis and distant metastasis of gastric cancer.Inhibition of PLCE1 gene can reduce the invasion and cell invasion and migration of gastric cancer SGC7901 cells,and its role mechanism may be to inhibit the EMT by inhibiting the PKC/ERK signaling pathway.