RESUMO
[Objective]This study was conducted to examine the effects of dexmedetomidine on the proliferation and angiogenesis of MHCC97H and SMCC7721 human hepatocellular carcinoma(HCC)cell lines cultured in hypoxia condition in vitro,and investigated the possible mechanism involved.[Methods]MHCC97H and SMCC7721 human HCC cell lines under hypoxia culture condition were treated with presence or absence of dexmedetomidine(100 μmol/L). Cell viability,colony formation,vasculogenic mimicry(VM) formation were assessed. The effects of dexmedetomidine on α-2A adrenergic receptor(α2A),hypoxia induced factor-1a(HIF-1a),and vascular endothelial growth factor(VEGF)protein expression were evaluated with Western blot analysis.[Results]Cell proliferation assay and colony formation assay indicated that hypoxia obviously promoted the proliferation of MHCC 97H and SMCC7721 cells(CoCl2 group vs corresponding control group,the proliferation rate of MHCC97H and SMCC7721:Day 3,142.2%and 133.8%;Day 4,134.7%and 131.0%;Day 5,133.5%and 136.2%;all P<0.05),and VM formation assay suggested that hypoxia increased angiogenesis of MHCC97H and SMCC7721 cells. Whereas dexmedetomidine significantly inhibited the proliferation(Dex+CoCl2 group vs CoCl2 group,the proliferation rate of MHCC97H and SMCC7721:Day 3,55.7%vs 60.7%;Day 4,46.9%vs 58.1%;Day 5,46.4%vs 57.0%,all P<0.05)and angiogenesis of MHCC97H,SMCC7721 cells induced by hypoxia. Dexmedetomidine may exert these functions by activating α-2A adrenergic receptor,causing an decrease in HIF-1a and VEGF protein,while hypoxia activated HIF-1a and VEGF protein to promote the growth and angiogenesis of cells.[Conclusion]The findings provide evidence that hypoxia could promote the proliferation and angiogenesis of MHCC97H and SMCC7721 cells,while dexmedetomidine might inhibit these effects by down-regulating HIF-1a and VEGF protein expression through activatingα-2A adrenergic receptor.
RESUMO
Aim To investigate the effect of neferine on proliferation and invasion of human hepatocellular car-cinoma. Methods HepG2 and Bel-7402 cells were cultured in vitro with different concentrations of nefer-ine, then cell proliferation was observed by CCK-8 as-say; cell invasion was observed by transwell invasion assay; the protein expression of RhoA, RhoC and ROCK was detected by Western blot. Results CCK-8 results showed that neferine could significantly inhibit cell proliferation in a dose-dependent manner compared with the control group. Transwell invasion assay showed that cell invasion was significantly decreased with neferine 3 μmol · L-1 . Western blot results showed that RhoA, RhoC and ROCK protein expres-sion was decreased when neferine was co-incubated with hepatocellular carcinoma cells. Conclusion Nef-erine can inhibit proliferation and invasion of HepG2 and Bel-7402 cells, which is mediated mainly by the inhibition of RhoA, RhoC and ROCK protein expres-sion.
RESUMO
Objective The cancer risk of patients with diabetes mellitus who are treated by metformin declines remarkably in comparison to patients receiving other drug therapies.The article was to investigate the relationship between antineopastic activity and fatty acid synthase (FASN) of metformin in human hepatocellular carcinoma cell(HCC) line HepG2. Methods HepG2 cells were treated with various concentrations of metformin( 0, 1, 5, 10, 15 mmol/L) for 24, 48 and 72 h respectively and cell growth was assessed by CCK-8 assay.Positive control(paclitaxel 10μg/mL) and negative control(metformin 0mmol/L) were set up simultaneously.After being treated with doses of metformin(0, 5, 10,15mmol/L) for 72h, protein expression levels of AMPKα、P-AMPKα、FASN、P-mTOR and P-Akt were measured by western blotting analysis and FASN mRNA expression levels were measured by RT-PCR. Results Being treated with vari-ous doses of metformin(1, 5, 10, 15 mmol/L) for 24, 48 and 72 h, the growth of HepG2 cells were inhibited by metformin in dose-dependent and time-dependent manner( P0.05) .FASN mRNA expression levels decreased significantly in all metformin-treated groups( P<0.05) . Conclusion Met-formin actitiviates AMPK, inhibits mTOR and downregulates FASN, which are implicated in its antineopastic activity on HCC.Although metformin inhibits mTOR activation, it is not involved in Akt upregulation through a negative loop.
RESUMO
Objective:To establish proteomic technique system of human hepatocellular carcinoma cell line QCY-7701. Method:Two different lysis buffer were taken to extract cell proteins. After two-dimensional gel electrophuresis (2-DE) and PDQuest analysis, 10 good-matched protein spots were chosen to be identified by MALDI-TOF-MS. Results:A steady 2-DE electrophregram of hepatocellular carcinoma cell line QGY-7701 was established. Compared with lysis bufferⅠ,protein quantities extracted from lysis bufferⅡincreased by 25%;protein spots in 2-DE gel increased by 32%. 9 out of 10 candidate protein spots were successfully identified (P
RESUMO
In the present study, the influence of essential free fatty acids on AFP secretion and cell growth of BEL-7402 human hepatocellular cacinoma cell line was investigated by radiommunoassay. The results demonstrated that 40 - 50?g/ ml concentration of linolenic acid could inhibit the AFP secretion obviously( P 0.05) . All of these studies about anti-cancer effect of linolenic acid will provide the principle for the patient for health care and therapy.
RESUMO
We used retroviral vector pLXSN to construct recombinant retroviral vectors with the human apoptosis gene, interleukin-l? converting enzyme (ICE). The vectors were introduced into packaging cell line PA317 by electroporation method. The G418 resistant colonies were selected, and the supematants of the colonies were used to infect the human hepatocellular carcinoma cell line SMMC7721. G418 resistant colonies of SMMC7721 were named SMMC7721-MCE and SMMC7721-neo. The results of RT-PCR analysis showed that exogenous hICE gene had successfully integrated into the genome of SMMC7721-hICE cells. The proliferation rate and tumorigenicity of cells in nude mice were examined. Our data showed that the growth rate and the tumorigenicity of SMMC7721-hICE cells in nude mice were considerablely decreased comparing with parent SMMC7721 and SMMC7721-neo. These results suggested that the retroviral vector expressing hICE gene was successfully constructed and could suppress the growth ability and tumorigenicity of human hepatocellular carcinoma cells, which provided a basis for further investigation of hICE gene therapy.
RESUMO
The effects of dexamethasone(Dex)on the 3H-Dex specific binding were studied in the human heptatocellular carcinoma cell line(SMMC-7721) by using counting vial culture assay.Cells were cultivated in the counting vials, as th; grew well and the monolayer cells adhered to the wall of vials.The cells were cultivated in the mdium containing 10-3,10-7, and 10-6mol/L Dex for 6, 12, 24 and 48 h, and then the specific binding of 3H-Dex at 30nmol/L of the ligand was determined after elimination of the effect of the occupancy of GR.It was found that 3H-Dex specific binding was not changed within 6 h, but decreased to 70% at 12 h and maintained at this level from 12 to 48 h in the Dex pretreated cells.The specific binding was recovered approximately to the control level 24 h after removal of Dex.These results indicate that glucocorticoids may down-regulate glucocorticoid receptor in vitro.