| Aim To study the expression of zinc transporter Zrt-and Irt-like proteins14(ZIP14)in the hepatocellular carcinoma(HCC)tissues,and investigate the effects of exogenous zinc and the overexpression of ZIP14 on the biological behavior of HCC cells.Methods 1.zip14 m RNA expression of the HCC tissues and adjacent non-tumor tissues from 81 HCC patients was detected with q RT-PCR method.ZIP14 protein expression of the HCC tissues and adjacent non-tumor tissues from 60 HCC patients was detected with immunohistochemical staining method.2.ICP-OES was used to detect the serum zinc levels of the patients with HCC,cirrhosis,chronic hepatitis and the healthy population.3.The effects of various concentration of zinc to the content and cellular distribution of zinc,cell proliferation,migration and invasion abilities,cell cycle and apoptosis of BEL-7404 cells with Zinc sulfate used as exogenous stimuli were detected by the TSQ fluorescent probe,MTT assay,transwell assay,DNA ploid and AO/EB fluorescence staining,respectively.4.The effects of various concentrations of zinc to the expressions of zip14,alb,bax and bcl-2 at m RNA levels in BEL-7404 cells were detected by q RT-PCR method.5.The lentivirus containing ZIP14 gene was constructed,and was used to infect the HCC cell line BEL-7404,which had relatively lower expression of ZIP14.Then the constructed HCC cell model BEL7404-ZIP14 was used to reveal the effects of ZIP14 overexpression on cell proliferation,migration and invasion abilities,and cell cycle of HCC cell,respectively.Results 1.The expression of zip14 at m RNA and protein levels of ZIP14 in HCC tissues were significantly lower than that of adjacent non-tumor liver tissues.Compared to other HCC cell lines,the expression of zip14 at m RNA in HCC cell line BEL-7404 was the lowest.2.The serum zinc levels of the patients with HCC and cirrhosis were significantly lower than that of the patients with chronic hepatitis and the health people,respectively(P<0.05,P<0.01).There was no significant difference in serum zinc levels between HCC patients and cirrhosis patients(P?0.05).3.With the increase of zinc concentration in culture conditions,the zinc level in BEL-7404 cell line was increased,the proliferation,migration and invasion ability of HCC cells were decreased,the cells were blocked in G2/M phases,and the apoptotic cells were increased.The expression of alb at m RNA levels in BEL-7404 cells exposed to 100 ?M zinc was increased(P<0.05)while the cellular expression of zip14 at m RNA level presented no changed with the concentration of zinc that stimulated the cells.4.The HCC cell model BEL7404-ZIP14 with overexpression of ZIP14 was constructed successfully.The cell proliferation,migration and invasion abilities of BEL7404-ZIP14 were inhibited compared with that of control group transfected with empty vector,the cell percentage in G2/M phase was increased(P<0.05),and the expression of alb was higher.The phenomena were more obvious when zinc concentration was increased in the culture condition.The m RNA expression of bcl-2 was remarkably reduced,while the m RNA expression of bax was increased than that of the control group(P<0.01).Conclusions 1.Both of the expressions of ZIP14 at m RNA and protein levels in HCC tissues are low,and the serum zinc concentrations of patients with HCC are significantly decreased.2.The high concentration of zinc might increase the intracellular zinc concentration of BEL-7404,inhibit the HCC cell proliferation and metastasis,block the cell cycle in G2/M phase,induce the cell apoptosis,and increase the alb m RNA expression.3.The overexpression of ZIP14 may facilitate the HCC cells to transfer zinc ion,and increase the expression of alb m RNA,inhibit the cell proliferation and metastasis,block the cell cycle in G2/M phase,and induce the cell apoptosis.The phenomina are especially obvious when the HCC cells are treated with exogenous zinc. |
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