| Objectives(1) The aim of this study was to compare the protein and m RNA levels of omentin-1 in human colon carcinoma tissues and para-carcinoma tissues. It was to prove the colon cancer cells could secrete and express the adipokine omentin-1.(2) The aim of this study was to identify the SW480 colon cancer cells could secrete and express the adipokine omentin-1 endogenously.(3) The aim of this study was to compare the protein and m RNA levels of p-Akt, TNF-α, IL-1β, VEGF-A in human colon carcinoma tissues and para-carcinoma tissues. Thereby, it was to investigate the adipokine omentin-1 could result in tumor progression via promotion of Akt signaling. Furthermore, it was aimed to analyze the relationships between protein expression of omentin-1 and TNF-α, IL-1β, VEGF-A and to explore the role of TNF-α, IL-1β, VEGF-A in tumorigenesis and developing of colon cancer.Methods:(1) The protein and m RNA expression levels of omentin-1 in human colon carcinoma tissues and para-carcinoma tissues were detected by q RT-PCR, Western blot and immunohistochemistry(IHC).(2) Human SW480 colon epithelial cancer cells were cultured. The supernatant and lyaste of SW480 cells were obtaibed from 6h, 12 h, 24 h and 48 h. Omentin-1 expression was investigated in SW480 colon cancer cells by q RT-PCR at m RNA level and confirmed by ELASA at protein level.(3) The protein expression levels of p-Akt and Akt in human colon carcinoma tissues and para-carcinoma tissues were detected by Western blot and the correlation among protein expression of omentin-1 and p-Akt/Akt in human colon carcinoma tissues was analyzed by Pearson correlation analysis and linear regression analysis method.(4) The protein expression levels of TNF-α, IL-1β, VEGF-A in human colon carcinoma tissues and para-carcinoma tissues were detected by IHC and the correlation among protein expression of omentin-1 and TNF-α, IL-1β, VEGF-A in human colon carcinoma tissues was analyzed by Pearson correlation analysis.Results:(1) The protein and m RNA levels of omentin-1 in human colon carcinoma tissues were significantly higher than those in para-carcinoma tissues determined by q RT-PCR, Western blot(m RNA: 0.983±0.105 vs 5.381±1.250, P=0.014<0.05, protein:1.086±0.242 vs 0.612±0.064, P<0.01). The positive expression of omentin-1 was detected in the cytoplasm and intercellular substance of human colon cancer cells by IHC, the expression of omentin-1 gradually reduced in para- cancer cells and colon epithelial cells. Moreover, the optical density value of human colon carcinoma tissues compared with para-carcinoma tissues was significantly increased by IHC(0.621±0.102 vs 0.295±0.030, P<0.01).(2) Omention-1 showed a high expression at both m RNA and protein levels in SW480 colon cancer cells. The concentration of this adipokine was greater in cell lysate compared with supernatants and secretion of omentin-1 was increased over time.(3) The protein expression levels of active p-Akt in human colon carcinoma tissues were significantly higher than those in para-carcinoma tissues determined by Western blot(1.891±0.245 vs 1.076±0.094, P=0.006<0.01). There was a significant positive correlation between the protein expression of omentin-1 and that of active p-Akt by Pearson correlation analysis(r=0.764,P<0.01). Linear regression analysis analysis showed that omentin-1 was the influencing factor of active p-Akt in colon cancer.(4) The protein expression levels of TNF-α, IL-1β and VEGF-A in human colon carcinoma tissues were significantly higher than those in para-carcinoma tissues determined by IHC(0.546±0.078 vs 0.231±0.064, 0.673±0.103 vs 0.336±0.044, 0.578±0.079 vs 0.250 ± 0.079, respectively, P < 0.01~0.05). Pearson correlation analysis showed that omentin-1 was negatively correlated with TNF-α 、 IL-1β(r=-0.767 and-0.334, respectively, P<0.05). However, there was no correlation between omentin-1 and VEGF-A in human colon carcinoma tissues.Conclusion:(1) These findings indicate the SW480 colon cancer cells could secrete and express the adipokine omentin-1 endogenously. Omentin-1 plays its possible carcinogenesis role in colon cancer through endocrine, autocrine and paracrine manner.(2) These findings indicate omentin-1 may affect the occurrence and progression of colon cancer via activation of Akt signaling.(3) The adipokine omentin-1 may not responsible for tumor microenvironment shaped by chronic inflammation state in the occurrence and progression of colon cancer. |
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