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Objective: filamin A (filamin A, FLNa) belonging to non-muscle actin-binding protein, is a major distribution of macromolecular proteins in the cytoplasm, initially thought to actin bundling, and can induce forming a strong three-dimensional actin gel form the cytoskeleton. Studies have shown that filamin A showed low levels of expression in colorectal cancer cells showed a high level of expression in normal tissue. FLNa expression level and TNM stage, liver metastasis, lymph node metastasis, tumor invasion depth correlation, but with the patient's age, gender, lesion, tumor size, tumor differentiation degree, general and pathological types do not have relevance. This research aimed at the FLNa the cloned into the vector pcDNA3.1/V5-His-TOPO liposome transfection of human colon cancer cells SW480 observed nude mice. Method: First this experiment reagent preparation, and then perform cell culture. FLNa gene plasmid vector pcDNA3.1/V5-His- TOPO / FLNa liposome-mediated transfection of these SW480 cells, in this experiment, we first determine the concentration of G418 selection. The the G418 concentration sieve to determine after the election of the three groups of cells, FLNa gene expression SW480 / FLNa cell. The two rounds of methods using reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot (Western blot) confirmed FLNa better expression of the transfected tumor cells. At the same time, we get a the stable expression FLNa gene SW480/FLNa cells. The three groups SW480 cell proliferative capacity in vitro by MTT assay, and incubation time for the horizontal axis (X-axis), the absorbance value of the vertical axis (Y-axis), the cell growth curve. The first 30 mice were randomly divided into three groups. Then SW480/FLNa, SW480 / pcDNA3.1 and SW480 three groups of cells were inoculated subcutaneously in nude mice, and tumor growth were observed; eagerly sacrificed all in nude mice after eight weeks, complete peel tumor, weighing, and analysis of tumor weight inhibitory rate was calculated; made transparent, fixed, dehydrated dip wax-embedded, and dewaxing treatment tumor tissue slices, then after HE staining, tumor tissue in the microscope morphological changes. Finally, the application SPSS13.0 statistical software for data analysis and processing. And the data were analyzed by t-test, one-way ANOVA, Dunnett-t test was used for the relevant inspection. Set test level α = 0.05, P lt; 0.05 for the difference was statistically significant. Results: 1 by using the RT-PCR method to detect the results of the three groups SW480 cells: the SW480 / mRNA expression levels of FL Na, SW480 / pcDNA3.1 and SW480 cells were 1.27 ± 0.11,0.12 ± 0.02 and 0.14 ± 0.03. SW480/FLNa group SW480 group (P = 0.019 lt; 0.05) the SW480/pcDNA3.1 group with SW480 difference between the groups was not statistically significant (P = 0.865 gt; 0.05); while using the Western blot assay results show protein expression levels SW480/FLNa, SW480/pcDNA3.1 and SW480 cells were 0.78 ± 0.03,0.01 ± 0.00 and 0.01 ± 0.00. SW480/FLNa group than SW480 group (P = 0.008), no statistically significant difference between SW480/pcDNA3.1 group and SW480 groups of (P = 0.943). Therefore, it can be seen that the FLNa gene in the SW480 cells i.e. (SW480/FLNa group) obtained in a relatively stable expression. 2 after the results of the MTT assay showed that the three groups of cells in 24h absorbance value of the difference was not statistically significant, the cell growth curve overlap, so the three groups of cell proliferative capacity in vitro. Through experiments that SW480/FLNa group is slow compared to the SW480/pcDNA3.1 and SW480 group tumor growth rate and tumor weight of SW480/FLNa group below SW480 group experimental statistical data show: [(0.67 ± 0.06) g vs (5.90 ± 0.77) g, P = 0.000]; inhibition was 88.7 ± 3.5% (P = 0.000); 4 after microscopy found, SW480/FLNa group of tumor tissue degeneration necrosis appears. Conclusion: The the FLNa gene transfection of human colon adenocarcinoma SW480 cells untransfected growth in nude mice compared with the gene of two groups of SW480 cells slowly. After HE staining in the light microscope display, SW480/pcDNA3.1 and SW480 tumor in nude mice were nests tumor cell atypia, nuclear chromatin deep, more visible split phase; SW480 / FLNa group tumor necrosis serious degeneration. In summary, the FLNa gene suppression SW480 cells, suggesting that FLNa gene has a role in suppressing tumor formation in nude mice. Further study on its mechanism for suppressing tumor can provide new ideas for the treatment of tumors.
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