| Objective: Observe the effects of sorafenib on HCC xenograft growth and detect differentially expressed mi RNA in tumor tissue. Analysis and discuss mi RNA involve in regulate some biological processes about inhibition of cancer cells proliferation., anti-angiogenic and epithelial to mesenchymal transition(EMT) after sorafenib intervention.Methods:1 To establish a model of Intraperitoneal xenografts in nude:Choose liver cell line MHCC-97 H which has high metastasis potential, and set up a model of subcutaneous tumor-bearing, when the tumor grow to about 1.5cm3, remove the tumor and cut it into small piece whose diameter is 3mm for the experiment. The nude mice for experiment was divided into group A(experimental group) and B(control group), each group of six. Then transplant the tumor tissue 1 piece in each nude mice to estabolish Intraperitoneal xenografts model. The mice was given 30 mg/kg(0.2ml) sorafenib, once daily. In contrast, the control received an equal volume of solvent, put to death all the mice after 35 days and draw the matierials.2 Compare the size of the tumor in the treatment group used sorafenib and the control, use immunohistochemical to detect tumor microvascular density(MVD) and hypoxia, stain hypoxia-induced factor-1α(HIF-1α), and detect the changes of E-cadherin and Vimentin of the tumor in the the experimental group and the control.3 Use Microarray to detect the differentially expressed mi RNA in two group, and bioinformatics analysis of its results. Analyze the molecular function of target genes using Gene Ontology(GO). Analyze the significant signaling pathways of the targetgene using Kyoko Encyclopedia of Genes and Genomes(KEGG).4 By means of Bioinformatics results, exploring mi RNA involved in proliferation, anti-angiogenic and EMT under sorafenib intervention, and to discuss the regulatory mechanism.Results:1 The tumor formation rate in nude mice were 100% in group A and B, no obvious gross tumor metastasis,and no mice died during the experiment.2 Tumor volume was significantly higher in control, MVD in experimental group was obvious lower expression than that in the control group, and the difference had statistical significance(P < 0.05). Expression of HIF-1α in experimental group was higher than the control, comparison of the two groups have statistical difference(P<0.05). Compared with the control group, the expression of E-cadherin in the experimental group was obviously down-regulated, and up-regulation of Vimentin expression, compared the two groups had significant difference(P<0.05), which suggested that sorafenib promote the process of EMT..3 Microarray shows that there was 28 mi RNAs differentially expressed in the two group, 20 of which were increased, and 8 of which were decreased. Through the results of bioinformatics analysis we concluded: mi R-762 and mi R-150-5p were upexpressed, which participated the process of sorafenib inhibiting hepatoma cells proliferation. Although mi R-142-a-3p was not increased obviously, while it was associated with many biological processes about tumor angiogenesis; mi R-155-5p increased and mi R-122-5p decreased participated in regulating the EMT process.Conclusion:1 Sorafenib equipped with the dual function: inhibit tumor growth and resistance to blood vessel formation, while it lead to tumor hypoxia, which indicated that it could promote EMT.2 Mi R-762 and mi R-150-5p participated in the process of regulation of sorafenib inhibit the growth of liver cancer; Mi R-142-a-3p was participated in the process of regulating tumor angiogenesis.3 Mi R-155-5p and mi R-122-5p participate in regulating the process of sorafenib induced EMT. |
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