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Objective To observe the effects of sevoflurane pretreatment on lung metastasis of mouse Lewis lung cancer (LLC)cells.Methods Mouse LLC cells were inoculated in culture plate. After being cultured for 24 h the cells were randomly divided into four groups:group control (CC), group 1% sevoflurane (SC1),group 2% sevoflurane (SC2),and group 3% sevoflurane (SC3).Cells of group SC1-3 were exposed to 1%,2%,3% sevoflurane for 4 h respectively,cells of group CC were exposed to 95%O 2-5%CO 2 mixture air,and were then cultured for another 24 h.The invasive activity of cells was determined by Transwell assay.The migration of cells was evaluated by wound scratch assay.The expression of MMP-2 and MMP-9 in cells were detected by ELISA.Thirty-two C57BL/6 mice were divided into four groups (n = 8):group control (CM),group 1% sevoflurane (SM1),group 2% sevoflurane (SM2),and group 3% sevoflurane (SM3).LLC cells of group SC1-3 were injected into caudal vein of mouse in group SM1-3 respectively.Cells of group CC were injected into mouse of group CM.Lung metastasis inhibitory rates were evaluated after 3 weeks. Results Compared with group CC,the invasive activity and migration of cells in group SC1-3 were decreased significantly,group SC1 >group SC2 >group SC3 (P group SC2>group SC3 (P <0.05).Compared with group CM,lung metastasis inhibitory rates of group SM1-3 were increased significantly,group SM1 < group SM2 < group SM3 (P < 0.05 ). Conclusion Sevoflurane can inhibit the lung metastasis of mouse LLC cells,which maybe through down-regulating the expression of MMP-2 and MMP-9 in mouse LLC cells.
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OBJECITVE: To study the inhibitive effect of plumbagin on Lewis lung cancer. METHODS: Cell proliferation was determined by CCK8 assay. Apoptosis was determined by flow cytometry. The expression of Bcl-2 and VEGF protein was studied by Western blot assay. The model of C57BL/6 mice bearing Lewis lung cancer was established by subcutaneous seeding of Lewis lung cancer cells, and randomly divided into 5 groups (n = 6). Tumor-bearing mice were injected with normal saline, plumbagin(low, medium, high dose) or cyclophosphamide (CTX) in each group. The tumor volume was measured. All mice were sacrificed on Day 22nd under aseptic condition for the tumor collection. The transplanted tumors were weighed for calculation of the tumor inhibition rate; Immunohistochemical method was applied to assessing the VEGF expression in tumor tissue. RESULTS CCK-8 assay showed that plumbagin had an obvious inhibition on Lewis lung carcinoma cells line in a dose-dependent manner(r = 0.953, P < 0.05). Plumbagin significantly increased cell apoptosis rate(P<0.05). The protein levels of Bcl-2 and VEGF were significantly reduced by plumbagin (0, 2.5, 5, 10 μmol·L-1) treatment(P < 0.05). In plumbagin(low, medium, high dose) groups and CTX group, the tumour volume, tumour weight and the expression of VEGF were significantly less than those in the control group (P < 0. 05). CONCLUSION: The plumbagin effectively inhibits Lewis lung carcinoma cells proliferation and tumor growth of Lewis lung carcinoma cells in mice. The mechanism involved is down-regulating the expression of Bcl-2, VEGF and inducing cell apoptosis.
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ATP-binding cassette protein E (ABCE1) has been annotated as an Rnase L inhibitor in eukaryotes. Previous study showed that the overexpression of ABCE1 was related with the occurrence and clinical stage of lung adenocarcinoma. As an initial investigation into the novel functions of ABCE1, siRNA-expressing vectors targeting sites of the ABCE1 gene were constructed from RNAi-Ready pSIREN-DNR-DsRed-Express vector. Cultured 95-D and NCI-H446 lung carcinoma cells were transfected with the siRNA-expressing vectors using FuGENE 6 and transfection efficiency was determined by using fluorescence microscopy. The expression level of ABCE1 protein was determined by Western blot and immunofluorescence staining. Cell viability was determined by MTT, cell cycle was analysed by flow cytometry.The apoptotic rate was observed by ELISA. Fluorescence microscopy showed a satisfactory transfection efficiency which was about 42.70%. Cell viability and the growth fraction were markedly suppressed, whereas the apoptosis was significantly increased in SiRNA-95-D and SiRNA-NCI-H446 cells than controls(P< 0.05). It can be concluded that the siRNA targeting ABCE1 gene shows a dramatic inhibitory effect on RNA transcription and protein expression and a promoting effect on the apoptosis in 95-D/NCI-H446 cells, which offers a reliable base for the further in vivo experiment.
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Objective To explore the effect of hypoxia on the in vitro invasion of lung carcinoma cells and its molecular basis. Methods Small cell lung carcinoma cells(H128) were exposed to hypoxia(5%O 2,5%CO 2,90%N 2) or anoxia(95%N 2,5%CO 2)condition for 48 hours. The invasive ability of the cancer cells was assayed with HABM-HEM model. The expression of E-cadherin and ? 1-integrin on the cells was also assayed by flow cytometry. Results Compared with normoxia group, the invasive ability of the cancer cells increased, the expression of E-cadherin decreased, and the expression of ? 1-integrin increased in hypoxia group. In anoxia group, the invasive ability and the expression of E-cadherin and ? 1-integrin all decreased compared with normoxia group. Conclusions Moderate hypoxia can down-regulate the expression of E-cadherin, up-regulate the expression of ? 1-integrin, and increase the invasion of carcinoma cells. But serious hypoxia can decrease the expression of adhesive molecules and the invasion of the cancer cells.