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AIM: To investigate the expression and effects of sialyl Lewis X (SLeX) on the invasion and migration of human hepatocellular carcinoma HepG2 cells.METHODS: The expression of α1,3-fucosyltransferase VII (FUT7) in HepG2 cells and L-02 cells was detected by RT-qPCR and Western blot.The SLeX expression in HepG2 cells and L-02 cells was determined by Western blot and immunocytochemical staining.The invasion and migration abilities of the treated cells were evaluated by Transwell assay.RESULTS: The expression of FUT7 and SLeX in the HepG2 cells, but not in the L-02 cells, was observed.The invasion rates of the HepG2 cells treated with SLeX monoclonal antibody at 0.05, 0.5 and 5 mg/L were significantly decreased as compared with control group (P<0.05).The migration ability of the HepG2 cells treated with SLeX monoclonal antibody at 0.05, 0.5 and 5 mg/L was also significantly reduced as compared with control group (P<0.05).The invasion rate and migratory cell number were significantly different between any 2 groups in the HepG2 cells treated with SLeX monoclonal antibody at 0.05, 0.5 and 5 mg/L (P<0.05).CONCLUSION: HepG2 cells express SLeX.SLeX is closely related to the migration and invasion abilities of the HepG2 cells.
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Objective:To investigate the mechanisms of IL-1β promoted lung cancer cells proliferation. Methods: The“Transwell? Inserts” system was used to coculture lung cancer cells A549,NCI-H520 with macrophages. BrdU ELISA used to measure the effect of macrophages promoted lung cancer cells proliferation. Expression of mRNA of IL-1β in A549 and NCI-H520 cells were analysed by Real-time PCR analysis. IL-1β was responsible for macrophage-promoted lung cancer cells growth, IL-1β neutralizing antibody was added. The autophagy marker Beclin1 protein was detected by Western blot. Results:The BrdU ELISA assay showed that after coincubation with macrophages in the proportion of 1:0. 5,the OD value of A549 increased from(0. 41±0. 06)to(1. 13±0. 10). There was statistical significance(P<0. 05). It also showed that the growth of the A549 cell was dependent on the macrophage number (P<0. 05). The OD value variability of NCI-H520 cells was as same as A549 cell upon cocultured with macrophages. Real-time PCR results showed that the expression of IL-1β mRNA in macrophages was remarkably enhanced in a time dependent manner upon coincubated with lung cancer cell,and the expression level was higher than lung cancer cells. Addition of IL-1β neutralizing antibody markedly inhibited macrophage-promoted lung cancer cells proliferation. The OD value of these two cells were decreased from ( 3. 63 ± 0. 33) to (1. 46±0. 18),from (2. 94±0. 38) to (1. 53±0. 20),respectively (P<0. 05). After treatment with IL-1β,the expression of Beclin1 was significantly inhibited in tumor cells. Conclusion:Over-expression of IL-1βfrom macrophages and lung cancer cells is re-sponsible for proliferation of tumor cells in coculture condition. Inhibition of autophagy in tumor cells may be the important mechanisms of IL-1β promotes lung cancer cells proliferation.
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Aim To study the inhibitory effects of gambogenic acid in combination with miR-218 on cervical cancer HeLa cells and its mechanisms.Methods Eukaryotic expression vector of miR-218(pmi8-218) was transfected into HeLa cells.Transcript levels of miR-218 were quantified by real-time quantitative PCR.HeLa cells were incubated with different concentrations of gambogenic acid alone or in combination with pmiR-218.The cell growth inhibiting ratio of HeLa cells was assessed by MTT assay.Cell apoptosis was measured by fluorescence activated cell sorting.The expression levels of Bcl-2, Bax and E-cadherin were measured by Western blot and qRT-PCR.Results Levels of miR-218 transcript significantly increased in pmiR-218 transfected HeLa cells.Overexpression of miR-218 may enhance the sensitivity of HeLa cells to gambogenic acid.Over expression of miR-218 could enhance the effect of gambogenic acid on inhibition cell proliferation, promoting apoptosis of HeLa cells.pmiR-218 could enhance the regulation of Bax expression and decrease the expression of Bcl-2 in HeLa cells.Conclusions Over expression of miR-218 may enhance the sensitivity of HeLa cells to gambogenic acid;miR-218 can enhance the effect of gambogenic acid on inhibition cell proliferation and promote the apoptosis of HeLa cells, and the mechanism may be related to down-regulation of Bcl-2/Bax expression.
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Objective To explore the effect and molecular mechanism of proteasome inhibitor in TNF-related apoptosis-inducing ligand (TRAIL)-induced apoptosis resistance on malignant lymphoma cells.Methods Raji cells were treated with TRAIL and proteasome inhibitor (PS-341) in vitro and the cell growth index was evaluated by MTT assay; cell cycle was analysed by flow cytometry; the protein and mRNA level of Bax were measured by Western blotting and real time RT-PCR. Results TRAIL inhibited proliferation of Raji cells at the concentration of 500 μg/L, but the inhibition rate was lower than that of the control cell:Hmy2.ciR.TRAIL arrested cell in G0/G1 phase. The Bax protein in Raji is degraded, but the Bax mRNA expression level does not change significantly .The effects of TRAIL was enhanced significantly 10 nmol/L PS-341 was added. Conclusion Raji cells are resistant in TRAIL-induced apoptosis. This effect may be related to the decrease of Bax protein. The Ubiquitin-proteasome pathway is involved in the degradation of Bax in TRAIL-treated Raji cells.
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Objective To explore whether antisense-EGFR could enhance the radiosonsitivity of human lung cancer spc-a-1 cell line.Methods The spc-a-1 cells were transfected with antisenso.EGFR-pcDNA3 by lipofectamine 2000(pcDNA3 antiEGFR group).Two other groups were used for comparison:control group(spc-a-1 cell without transfection)and pcDNA3 group(spc-a-1 cell transfeeted with pcDNA3 which did not contain antisense EGFR).Cell clones that stable expressing antisense-EGFR wa8 selected with G41 8 and the suppression of the expression of EGFR mRNA and protein were detected by RT-PCR and Western blot.The influence of antisense-EGFR on cell cycle was testified by flow cytometry assay.The cell apoptosis was analyzed by flow cytometry after 8 Gy irradiation.Further,cells of each group were irradiated with X-rays at the dose of 0,2,4,6 and 8 Gy.Dose-survival curve of each group was established by colony-forming assay.Results The expression of EGFR mRNA and protein were significantly inhibited after antisense-EGFR-pcDNA3 transfection.The cells arrested at the G2/M phase in the pcDNA3 antiEGFR group,control group and pcDNA3 group were (29.53±1.91)%,(13.7±1.30)%and(12.40±1.34)%,respectively.The apoptosis index of spc-a-1 cells in the antisonse-EGFR combined with irradiation group was obviously higher than that of the comparable groups [(39.24±1.57)%,(13.79±0.63)%and(15.02±0.85%)].The values of D0,Dq,SF2 of pcDNA3 antiEGFR group declined obviously compared with the control group(2.11,2.49,0.84 vs 1.19,0.15,0.32).Conclusions Antisense-EGFR could induce the G2/M cell cycle arrest,promote cell apoptosis and inhibit the ability of sublethal cell damage repair induced by irradiation,80 that it could significantly improve the radiosensitivity of spc-a-1 cell in vitro.
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Objective To reform experimental course teaching methods of clinical bio-chemistry and elevate students interesting and ability. Methods The reformed teaching methods was taken in clinical laboratory techniques and start case-guide comprehensive experiment of clinical biochemistry. Results The innovation of teaching methods of clinical biochemistry obtained satisfactory achievement,improved students thinking skills and overall quality. Conclusion It accorded with the trend of teaching innovation and was advantageous to increasing the comprehensive predisposition.
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Objective: To explore the inhibition effects of ectogenic wild type p53 cDNA(Ad wtp53) on colorectal carcinoma cell lines with different p53 gene status and search for the role of wild type p53 tumor suppressor gene in occurrenc and progress of malignant tumor. Methods: MTT process was taken to choose optimal transfection titre. Three kinds of cell lines(p53 gene deletion, mutation and nomal) were transferred by Ad wtp53 in optimal titre. The inhibition effects of these cell lines were observed and compared. Results: The best titre is 1000 MOI and p53 gene deletion cell line (THC 8908) shew the highest sensitivity. G 1 S transition period blocking effects occurred in all cell lines and G 2 M phase regulation effects were not coincidence in three colorectal cell lines. Conclusions: Recombinant adenovirus mediated wild type p53 gene observably inhibited colorectal carcinoma cell lines growth and proliferation, blocked cell cycle in G 0 /G 1 phase and displayed obvious different actions on G 2 M phase among cell lines with different p53 status.
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Objective To study the relationship of fatty acid binding protein-2(FABP2)and apolipoprotein E(ApoE)with coronary heart disease(CHD)in type-2 diabetes mellitus(T2DM)patients.Methods FABP2 and ApoE gene polymorphisms were detected by PCR-RFLP.At the same time all CHD in T2DM patients' serum triglyceride(TG),total cholesterol(TC),high-density lipoprotein cholesterol were(HDLC),low-density lipoprotein cholesterol(LDLC)were detected.Results The T-allele frequencies of FABP2 were higher(P