Abstract
To investigate the effects of lentiviral vector‐mediated shRNA suppressing CXCR7 on tumour invasion and metastasis in hepatocellular carcinoma (HCC) after transcatheter arterial chemoembolization (TACE). HCCLM3 cell lines were cultured and assigned into the CXCR7‐shRNA, negative control (NC) and blank groups. The qRT‐PCR and Western blotting were applied to detect the mRNA and protein expressions of CXCR7, CXCR4 and MMP‐2 in HCCLM3 cells. Cell proliferation and invasion were evaluated by MTT and Transwell assays. A Buffalo rat model of HCC was established. Fifty model rats were divided into the CXCR7‐shRNA + TACE, CXCR7‐shRNA, TACE, NC and control groups. Immunohistochemistry was performed to detect the expressions of CXCR7, MMP‐2, vascular endothelial growth factor (VEGF) and intratumoral CD31‐positive vessel count in tumour tissues of mice. Compared with the blank and NC groups, the mRNA and protein expressions of CXCR7 and MMP‐2 were decreased in the CXCR7‐shRNA group. The cell proliferation and invasion rates of the CXCR7‐shRNA group were lower than the blank and NC groups. At the 4th week after TACE, tumour weight of the CXCR7‐shRNA + TACE group increased continuously. The CXCR7‐shRNA + TACE group showed longer survival time and smaller tumour sizes than other groups. Compared with other groups, the CXCR7‐shRNA + TACE and CXCR7‐shRNA groups had less number of lung metastatic nodules and lower expressions of CXCR7, MMP‐2, VEGF and CD31‐positive vessel count. CXCR7‐shRNA inhibits tumour invasion and metastasis to improve the efficacy of TACE in HCC by reducing the expressions of CXCR7, MMP‐2 and VEGF.
Highlights
Primary liver cancer is the sixth most common cancer worldwide, and the third and sixth most common causes of cancer-related mortality [1, 2]
At 72 hrs after infection by LV-CXC chemokine receptor 7 (CXCR7)-short hairpin RNA (shRNA), the HCCLM3 cells were observed under an inverted fluorescence microscope for green fluorescent protein (GFP) expression, which was identified in the majority of the cells (Fig. 1), indicating that the cells grew well and LV-CXCR7-shRNA efficiently infected the HCCLM3 cells
We simultaneously measured the protein expressions of CXCR7, CXCR4 and MMP-2 in HCCLM3 cells of the three groups by Western blotting and identified similar findings that the CXCR7-shRNA group showed significantly lower protein expressions of CXCR7 and MMP-2 than the negative control (NC) group (P < 0.001); no differences were found in the protein expressions of CXCR7 and MMP-2 between the blank group and the NC group (P > 0.05)
Summary
Primary liver cancer is the sixth most common cancer worldwide, and the third and sixth most common causes of cancer-related mortality [1, 2]. One of the major challenges in improving the survival rate of patients with
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