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Objective To investigate the effect of microRNA-134-5p (miR-134-5p) targeting epidermal growth factor receptor (EGFR) on the growth of ovarian cancer cells.Methods The ovarian cancer cell lines SKOV3 and A2780 served as the study objects and were divided into the control group (transfecting miR-NC) and experimental group (transfecting miR-134-5p) according to the treatment method.The expression levels of EGFR gene and downstream target protein were detected by qRT-PCR and western blot.The cell cycle distribution and apoptosis were detected by flow cytometry.The proliferation ability of ovarian cancer cells was detected by MTT assay and colony forming assay.Results The expressions of EGFR and downstream target protein in the experimental group were significantly down-regulated.EGFR mRNA in SKOV3 cells was downregulated to 48％ (P＜0.05),and EGFR mRNA in A2780 cells was down-regulated to 47％ (P＜0.05).The cell cycle of cells in the experimental group was significantly inhibited (P＜0.05),and miR-134-5p induced apoptosis through the EGFR target protein (P＜0.05).The proliferation activity and colony forming ability of the experimental group were significantly inhibited (P＜0.05).Conclusion miR-134-5p could promote the cellular cycle arrest and apoptosis,and reduces the proliferation ability of ovarian cancer cells by targetedly inhibiting the EGFR gene.
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Objective To explore the correlation between Y-box binding protein-1 ( YB-1) and P-glycoprotein ( P-gp) in drug-resistant hepatocellular carcinoma ( HCC) Bel-7402/ADM cells, and speculate the related mechanism of drug resistance.Methods Bel-7402/ADM cells were developed by concentration gradient escalation and intermittent administration of large dose. The levels of YB-1 mRNA and MDR1 mRNA were detected by means of RT-PCR.Western blotting was used to detect the protein expression of YB-1 and P-gp in the Bel-7402 cells and doxorubicin resistant Bel-7402 ( Bel-7402/ADM) cells. Bel-7402/ADM cells were transfected with small interfering RNA ( siRNA) targeting human YB-1. Expression levels of YB-1 and MDR1 mRNA and protein were detected by means of RT-PCR and Western blotting. Results IC50 values of ADM on hepatoma carcinoma cells Bel-7402 and Bel-7402/ADM were (2.23±0.07) and (7.02±0.03) μmol?L-1. The mRNA expression levels of MDR1 and YB-1 were all significantly higher in Bel-7402/ADM cells than in Bel-7402 ( P<0.01) . The mRNA expression levels of MDR1 and YB-1 in Bel-7402/ADM cells transfected with YB-1 siRNA were reduced significantly (P<0.01). The protein levels of YB-1 and MDR1 in Bel-7402/ADM cells transfected with YB-1 siRNA were reduced significantly ( P<0. 05 ) . Conclusion These results suggest that the high expression level of YB-1 is probably correlated with multidrug resistance in HCC Bel-7402/ADM cells.
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Objective To explore the correlation between Y-box binding protein-1 ( YB-1) and P-glycoprotein ( P-gp) in drug-resistant hepatocellular carcinoma ( HCC) Bel-7402/ADM cells, and speculate the related mechanism of drug resistance.Methods Bel-7402/ADM cells were developed by concentration gradient escalation and intermittent administration of large dose. The levels of YB-1 mRNA and MDR1 mRNA were detected by means of RT-PCR.Western blotting was used to detect the protein expression of YB-1 and P-gp in the Bel-7402 cells and doxorubicin resistant Bel-7402 ( Bel-7402/ADM) cells. Bel-7402/ADM cells were transfected with small interfering RNA ( siRNA) targeting human YB-1. Expression levels of YB-1 and MDR1 mRNA and protein were detected by means of RT-PCR and Western blotting. Results IC50 values of ADM on hepatoma carcinoma cells Bel-7402 and Bel-7402/ADM were (2.23±0.07) and (7.02±0.03) μmol?L-1. The mRNA expression levels of MDR1 and YB-1 were all significantly higher in Bel-7402/ADM cells than in Bel-7402 ( P<0.01) . The mRNA expression levels of MDR1 and YB-1 in Bel-7402/ADM cells transfected with YB-1 siRNA were reduced significantly (P<0.01). The protein levels of YB-1 and MDR1 in Bel-7402/ADM cells transfected with YB-1 siRNA were reduced significantly ( P<0. 05 ) . Conclusion These results suggest that the high expression level of YB-1 is probably correlated with multidrug resistance in HCC Bel-7402/ADM cells.
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Hepatocellular carcinoma (HCC) is one of leading causes of death in the world. Although various treatments have been developed, the therapeutic side effects are far from desirable. Chinese medicines (CMs, including plants, animal parts and minerals) have drawn a great deal of attention in recent years for their potential in the treatment of HCC. Most studies have shown that CMs may be able to retard HCC progression with multiple actions, either alone or in combination with other conventional therapies to improve quality of life in HCC patients. Additionally, CMs are used for preventing HCC occurrence. The aim of this study is to review the potential prophylactic and curative effects of CMs on human HCC and the possible mechanisms that underlie these pharmacological actions. Publications were collected and reviewed from PubMed and China National Knowledge Infrastructure from 2000 to 2014. Keywords for literature searches include "Chinese medicine", "Chinese herb", "traditional Chinese Medicine", "hepatocellular carcinoma" and "liver cancer". CMs in forms of pure compounds, isolated fractions, and composite formulas are included. Combination therapies are also considered. Both in vitro and in vivo efficacies of CMs are being discussed and the translational potential to bedside is to be discussed with clinical cases, which show the actions of CMs on HCC may include tumor growth inhibition, antimetastatic activities, anti-inflammation, anti-liver cancer stem cells, reversal on multi-drug resistance and induction/reduction of oxidative stress. Multiple types of molecules are found to contribute in the above actions. The review paper indicated that CMs might have potential to both prevent HCC occurrence and retard HCC progression with several molecular targets involved.
Subject(s)
Humans , Carcinoma, Hepatocellular , Drug Therapy , Drug Resistance, Multiple , Liver Neoplasms , Drug Therapy , Medicine, Chinese Traditional , NF-E2-Related Factor 2 , Physiology , Neoplastic Stem Cells , Reactive Oxygen Species , MetabolismABSTRACT
Aims To study the effects of clenbuterol on anoxia/reoxygenation( A/R) injury in neonatal Wistar rat cardiomyocytes and to explore whether its mecha-nism is related to reperfusion injury salvage kinase ( RISK) or not. Methods The cultured primary neo-natal cardiomyocytes were randomly divided into eight groups: ①normal culture group; ②anoxia/reoxygen-ation( A/R) group;③ clenbuterol ( 1 μmol · L-1 ) +A/R;④ICI118,551(10 μmol·L-1) + clenbuterol ( 1 μmol · L-1 ) + A/R; ⑤Metoprolol ( 10μmol · L-1 ) + clenbuterol(1μmol·L-1 ) + A/R group;⑥Metoprolol ( 10 μmol · L-1 ) + A/R group; ⑦PD98059 ( 20 μmol · L-1 ) + clenbuterol ( 1 μmol · L-1 ) + A/R group;⑧ LY294002(10 μmol·L-1 ) +clenbuterol(1 μmol · L-1 ) + A/R group. Cell via-bility was determined by the conventional MTT reduc-tion assay. The content of LDH in cultured medium was measured with colorimetry. Cardiomyocyte apopto-sis was determined by Hoechst33342 . Intracellular re-active species( ROS) were monitored by the fluorescent DCFH-DA. Total ERK2 and phosphorylated ERK were detected by western blot. Results Compared with A/R group, clenbuterol significantly increased vaibility of cells, reduced LDH release, lowered the rate of apop-tosis and ROS production. When addedβ2 receptor an-tagonist ICI118 , 551 , PI3 K inhibitor LY294002 and ERK inhibitor PD98059 , the effects of clenbuterol a-bove were inhibited; but β1 receptor antagonist Meto-prolol protected the cardiomyocytes from A/R injury, as evidenced by decreased LDH release and increased cell viability. There were no synergistic effects in the combined use of clenbuterol and Metoprolol. Conclu-sion clenbuterol exerts cardioprotective effects against A/R injury by inhibiting oxidative stress and apopto-sis. The protection of clenbuterol is inhibited by ICI118 , 551 , LY294002 and PD98059 . clenbuterol protects cardiomyocytes against A/R injury via RISK pathway by activation of β2 receptor.
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Objective To establish a method for simultaneous determination of icariin and naringin content in bushen huayu extract by HPLC. Methods Reverse-phase high performance liquid chromatography ( RP-HPLC ) separation was performed one C18 column (4.6 mmí250 mm,5 μm).The mobile phase was acetonitrile containing 0.2%H4PO3(pH=2.9). Gradient elution with a flow rate of 0. 8 mL·min-1 was applied to achieve the separation. The detection wavelength was set at 269 nm,and the column temperature was 30℃. Results Icariin had a good linear range from 0. 3-3. 0μg (r=0. 999 9),the recovery rate was 105. 6%,and RSD was 0. 95%. Naringin was linear within the range of 0. 12-1. 20 μg (r=0. 999 9). The recovery rate was 94. 0%and RSD was 0. 52%. Conclusion The method is simple,stable,accurate,and reproducible,which can be used as the quality control standard for bushen huayue extract.