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Objective To investigate the relationship between the expression of XB130 and the histodifferentiation,lymph node metastasis and survival time of patients with gastric cancer.Methods Seventy-two specimens of gastric cancer tissues and seventy-two specimens of paracancerous tissues were collected from June 2011 to June 2012 in Henan Province People's Hospital.The expression of XB130 protein in gastric cancer and paracancerous tissues was detected by immunohistochemistry.The expression of XB130 mRNA in gastric cancer and paracancerous tissues was detected by real-time quantitative polymerase chain reaction.The relationship between the expression of XB130 and clinicopathological features of gastric cancer was analyzed.Results There were 26 cases of positive expression of XB130 protein and 46 cases of negative expression of XB130 protein among the 72 cases of gastric cancer tissues,and the positive expression rate was 36.1％ (26/72).There were 52 cases of positive expression of XB130 protein and 20 cases of negative expression of XB130 protein among the 72 cases of paracancerous tissues,and the positive expression rate was 72.2％ (52/72).The positive expression rate of XB130 protein in gastric cancer tissues was significantly lower than that in paracancerous tissues (x2 =16.200,P ＜ 0.05).The positive expression rate of XB130 protein in poorly and moderately differentiated gastric cancer tissues was significantly lower than that in well differentiated gastric cancer tissues (x2 =5.786,P ＜0.05).The positive expression rate of XB130 protein in gastric cancer tissues with lymph node metastasis was significantly lower than that in gastric cancer tissues without lymph node metastasis (x2 =4.281,P ＜0.05).The relative expression of XB130 mRNA in gastric cancer tissues and paracancerous tissues was 1.52 ±0.46 and 2.28-± 0.51 respectively,the expression of XB130 mRNA in gastric cancer tissues was significantly lower than that in paracancerous tissues (t =-21.744,P ＜0.05).The expression of XB130 mRNA in well differentiated gastric cancer tissues was significantly higher than that in poorly and moderately differentiated gastric cancer tissues (t =-13.982,P ＜ 0.05).The expression of XB130 mRNA in gastric cancer tissues with lymph node metastasis was significantly lower than that in gastric cancer tissues without lymph node metastasis (t =-19.906,P ＜ 0.05).The mean survival time in patients with high and low expression of XB130 mRNA was (37.040 ± 14.826) and (21.529 ± 11.789) months respectively,the survival time in patients with high expression of XB130 mRNA was significantly higher than that in patients with low expression of XB130 mRNA (t =9.121,P ＜0.05).Conclusion XB130 may be involved in the occurrence and development of gastric cancer,and it is associated with the differentiation,lymph node metastasis and survival time of patients with gastric cancer.
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<p><b>Background</b>XB130 is a recently discovered adaptor protein that is highly expressed in many malignant tumors, but few studies have investigated its role in hepatocellular carcinoma (HCC). Therefore, this study explored the relationship between this protein and liver cancer and investigated its molecular mechanism of action.</p><p><b>Methods</b>The expression of XB130 between HCC tissues and adjacent nontumor tissues was compared by real-time polymerase chain reaction, immunochemistry, and Western blotting. XB130 silencing was performed using small hairpin RNA. The effect of silencing XB130 was examined using Cell Counting Kit-8, colony assay, wound healing assay, and cell cycle analysis.</p><p><b>Results</b>We found that XB130 was highly expressed in HCC tissues (cancer tissues vs. adjacent tissues: 0.23 ± 0.02 vs. 0.17 ± 0.02, P < 0.05) and liver cancer cell lines, particularly MHCC97H and HepG2 (MHCC97H and HepG2 vs. normal liver cell line LO-2: 2.35 ± 0.26 and 2.04 ± 0.04 vs. 1.00 ± 0.04, respectively, all P < 0.05). The Cell Counting Kit-8 assay, colony formation assay, and xenograft model in nude mice showed that silencing XB130 inhibited cell proliferative ability both in vivo and in vitro, with flow cytometry demonstrating that the cells were arrested in the G0/G1 phase in HepG2 (HepG2 XB130-silenced group [shA] vs. HepG2 scramble group [NA]: 74.32 ± 5.86% vs. 60.21 ± 3.07%, P < 0.05) and that the number of G2/M phase cells was decreased (HepG2 shA vs. HepG2 NA: 8.06 ± 2.41% vs. 18.36 ± 4.42%, P < 0.05). Furthermore, the cell invasion and migration abilities were impaired, and the levels of the epithelial-mesenchymal transition-related indicators vimentin and N-cadherin were decreased, although the level of E-cadherin was increased after silencing XB130. Western blotting showed that the levels of phosphorylated phosphoinositide 3-kinase (PI3K) and phospho-protein kinase B (p-Akt) also increased, although the level of phosphorylated phosphatase and tensin homolog increased, indicating that XB130 activated the PI3K/Akt pathway. Furthermore, we found that a reduction in XB130 increased liver cancer cell sensitivity to tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis.</p><p><b>Conclusions</b>Our findings suggest that XB130 might be used as a predictor of liver cancer as well as one of the targets for its treatment.</p>
Subject(s)
Animals , Mice , Adaptor Proteins, Signal Transducing , Genetics , Metabolism , Apoptosis , Carcinoma, Hepatocellular , Metabolism , Pathology , Cell Movement , Cell Proliferation , Gene Knockdown Techniques , Liver Neoplasms , Metabolism , Pathology , Mice, Nude , Microfilament Proteins , Genetics , Metabolism , Neoplasm Invasiveness , Phosphatidylinositol 3-Kinases , Signal TransductionABSTRACT
Background and purpose: XB130 protein plays an important role in proliferation and invasiveness of tumor cells. However, there is little research on the role of XB130 protein in hepatocellular carcinoma (HCC) and the effect of XB130 is still unclear. This study investigated the role of XB130 gene in the proliferation of HCC cell and its potential mechanism. Methods: The protein expressions of XB130 in HCC cell lines, Huh7, HepG2 and SNU449, and liver cell line HL7702 were detected by Western blot. Huh7 cells were transfected with XB130-siRNA. Then cell viability was measured using cell counting kit-8 (CCK-8), and cell cycle was examined by flow cytometry. Protein expressions of p-AKT, p-GSK3β, cyclin D1 and p-Rb were detected by Western blot, while mRNA expression levels of E2F/DP1 target genes (cyclin E1, c-Myc and PCNA) were measured by reverse transcription-polymerase chain reaction (RT-PCR). Results: The relative protein expressions of XB130 in Huh7, HepG2, SNU449 and HL7702 cells were 0.66±0.10, 0.78±0.11, 0.83±0.08 and 0.32±0.06, respectively. The difference between HCC cell lines and HL7702 cell line was statistically significant (P＜0.01). The transfection efficacy of XB130-siRNA was confirmed to be highly effective in Huh7 cells, and the viability of XB130-siRNA transfected Huh7 cells declined 72 h after transfection (P＜0.001). The ratio of Huh7 cells in G0/G1 phase was increased, while the ratio in S or G2/M was decreased 48 h after XB130-siRNA transfection (P＜0.01). In addition, compared with negative control, protein expressions of p-AKT, p-GSK3β, cyclin D1 and p-Rb, and mRNA expression levels of cyclin E1, c-Myc and PCNA were all decreased in XB130-siRNA transfected Huh7 cells (P＜0.001). Conclusion: XB130 promotes the proliferation of HCC cells by regulating cell cycle-related proteins and downstream transcription factors.
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Currently,XB130 as a newly discovered characterized adaptor protein ,it has been implicated as a substrate and regulator of many intracellular signal transduction ,such as FAK/SRC,PI3K/Akt and MEK-ERK signaling and so on.It has been found that XB130 is high expression in many cell lines ,for instance thyroid carcinoma,osteosarcoma,gastric cancer,esophageal cancer and breast cancer etc .The mechanism of XB130 in tumor is becoming increasingly attention .XB130 is recently attributed to be a new oncogene ,and plays important roles in cell pro -liferation,cell survival and tumorigenesis .A deeper understanding of these mechanisms may lead to the discovery of XB130 as an important mediator in tumor development and as a novel therapeutic target for cancer.