Objective To investigate the role and implication of Engrailed-2 (EN2) in human hepatocellular carcinoma (HCC) and the effect of silencing EN2 genes on the proliferation and invasion of HepG2 cells by RNA interference.Methods Immunohistochemistry was used to detect the expression of EN2 in the HCC tissues and corresponding non-cancerous adjacent liver tissues in 126 patients with HCC.The expressions of EN2 and the relationship between EN2 expression and clinicopathological features and prognosis were analyzed using the Chi-square test.The survival curve was drawn using the Kaplan-Meier method and the survival was analyzed using the log-rank test.EN2-small interfering RNA (siRNA) was transfected into HepG2 cell lines mediated by LipofectamineTM 2000,and the expression of EN2 were detected by Western blotting assay.The cell proliferation and invasion were measured by methyl thiazol tetrazolium (MTT) and Transwell assays.Results EN2 positive expression rate was high (62.7％) in HCC tissue but low in the corresponding peritumoral tissue (23.5％,P ＜ 0.01).The high expression of EN2 was strongly correlated with tumor size,metastasis and AJCC TNM stage.The cumulative 5-year survival rate was 33.9％ in the low EN2 expression group,whereas it was 5.1％ in the high EN2 expression group (P ＜ 0.01).Expression of EN2 in EN2 siRNA group was significantly lower than that in control siRNA group and control group (P ＜ 0.05),respectively.The cell proliferation and invasion ability were significantly reduced in the EN2 siRNA group.Conclusion The expression of EN2 is highly up-regulated in HCC tissues and down-regulation of EN2 could inhibit proliferation and invasion of HepG2 cells,which indicating that EN2 is involved in the process of HCC carcinogenesis and progression and may serve as a biomarker for predicting prognosis of HCC patients.

This study examined the role of regulated upon activation normal T cell expressed and secreted (RANTES) and its receptor C-C chemokine receptor type 5 (CCR5) in gastric cancer metastasis and the associated mechanism. The expression of RANTES and CCR5 was detected by using immunohistochemical staining and Western blotting in the gastric cancer tissues obtained from 60 gastric cancer patients with or without lymph node metastasis (n=30 in each). The results showed that the expression levels of RANTES and CCR5 were higher in gastric cancer with lymph node metastasis than in that without metastasis (P<0.05). The expression levels of RANTES in 30 lymph nodes with cancerous invasion were higher than in 30 normal lymph nodes (P<0.05). Chemotactic test revealed that the number of migrating gastric cancer cells (n=295.0±54.6) induced by the protein of cancer-invading lymph nodes was greater than that by the protein mixture from cancer-invading lymph nodes and RANTES antibody (n=42.5±11.6) (P<0.05). RT-PCR showed that the expression levels of the main Th1 cytokines (IL-2, Γ-IFN) were lower in gastric cancer with lymph node metastasis (2.22±0.90, 3.26±1.15 respectively) than in that without metastasis (3.07±1.67, 4.77±1.52 respectively) (P<0.05), but the expression level of the main Th 2 cytokine (IL-10) was higher in gastric cancer with lymph nodes metastasis (6.06±2.04) than in that without metastasis (4.88±1.87) (P<0.05). It was concluded that RANTES and its receptor CCR5 may contribute to gastric cancer metastasis through influencing the balance of Th1/Th2. RANTES and CCR5 may become a marker of gastric cancer metastasis.

This study examined the role of regulated upon activation normal T cell expressed and secreted (RANTES) and its receptor C-C chemokine receptor type 5 (CCR5) in gastric cancer metastasis and the associated mechanism.The expression of RANTES and CCR5 was detected by using immunohistochemical staining and Western blotting in the gastric cancer tissues obtained from 60 gastric cancer patients with or without lymph node metastasis (n=30 in each).The results showed that the expression levels of RANTES and CCR5 were higher in gastric cancer with lymph node metastasis than in that without metastasis (P＜0.05).The expression levels of RANTES in 30 lymph nodes with cancerous invasion were higher than in 30 normal lymph nodes (P＜0.05).Chemotactic test revealed that the number of migrating gastric cancer cells (n=295.0±54.6) induced by the protein of cancer-invading lymph nodes was greater than that by the protein mixture from cancer-invading lymph nodes and RANTES antibody (n=42.5+11.6) (P＜0.05).RT-PCR showed that the expression levels of the main Th1 cytokines (IL-2,γ-IFN) were lower in gastric cancer with lymph node metastasis (2.22±0.90,3.26±1.15 respectively)than in that without metastasis (3.07±1.67,4.77±1.52 respectively) (P＜0.05),but the expression level of the main Th 2 cytokine (IL-10) was higher in gastric cancer with lymph nodes metastasis (6.06±2.04)than in that without metastasis (4.88±1.87) (P＜0.05).It was concluded that RANTES and its receptor CCR5 may contribute to gastric cancer metastasis through influencing the balance of Th1/Th2.RANTES and CCR5 may become a marker of gastric cancer metastasis.