ABSTRACT
Objective:To investigate the expression of circular RNA 102958 (circRNA_102958) in human gastric cancer tissues and its clinical significance.Methods:Thirty cancer tissues from gastric cancer patients in Jiangsu Cancer Hospital from September 2017 to March 2018 and their matched normal gastric mucosa tissue samples were collected. The relative expression of circRNA_102958 in the tissues was detected by fluorescence quantitative real-time polymerase chain reaction (qRT-PCR). The relationship between the expression of circRNA_102958 in gastric cancer tissues and clinicopathological features was analyzed. The receiver operating characteristic curve (ROC) was used to evaluate the efficacy of circRNA_102958 in the diagnosis of gastric cancer.Results:The relative expression of circRNA_102958 in gastric cancer tissues was higher than that in normal gastric mucosa tissues, and the difference was statistically significant (5.1±1.0 vs. 1.0±0.0, t = 4.045, P = 0.000 2). The relative expression of circRNA_102958 in gastric cancer tissues of patients with stage Ⅲ-Ⅳ was higher than that of patients with stage Ⅰ-Ⅱ, and the difference was statistically significant (9.3±2.6 vs. 2.0±0.5, t = 2.302, P = 0.029). There was no statistical difference in the relative expression of circRNA_102958 among patients with different gender, age, the longest tumor diameter, tissue differentiation degree, and lymph node metastasis (all P > 0.05). ROC analysis showed that the sensitivity and specificity of circRNA_102958 in the diagnosis of gastric cancer were 74% and 61%, and the area under the curve was 0.74. Conclusions:circRNA_102958 is highly expressed in gastric cancer tissues, and its expression is related to the stage of gastric cancer, which may be related to the occurrence and development of gastric cancer. circRNA_102958 is expected to become a molecular marker for gastric cancer diagnosis.
ABSTRACT
Objective To investigate the effect of exosomes secreted from human lung adenocarcinoma A549 cells under hypoxic or normoxic conditions on the radiosensitivity and invasiveness of normoxia cells.Methods A549 cells were cultured in hypoxic (1％ O2) and normoxic (21％ O2) conditions,respectively.The exosomes (N-EXO and H-EXO) secreted from normoxic or hypoxic A549 cells were collected by ultracentrifugation and its number was measured using a NanoSight detector.The appearance and size distribution of exosomes were observed by a scanning electron microscopy.The exosomal marker protein CD63 was measured by Western blot.The proliferation of cells exposed to X-rays under hypoxic or normoxic conditions were detected by CCK8 assay.The cell uptake situation of exosomes labeled with PKH67 was observed by a fluorescence microscopy.Cell migration and invasiveness were detected by a cell scratch test and transwell assay.The expression of matrix metalloproteinase 2 (MMP2) and MMP9 was detected by ELISA.Cellular radioresistance effect of exosomes was evaluated by a colony formation assay.Results The NanoSight measurement showed the number of exosomes in cell culture medium was increased after hypoxia treatment.The H-EXO and N-EXO showed typical ring cake shape.The size distribution of H-EXO was mainly between 30 nm and 200 nm,smaller than that of N-EXO (50-220 nm).Western blot assay showed that CD63 was expressed in both H-EXO and N-EXO.At 4 and 6 days after 2 Gy X-rays irradiation,cell proliferation rate of hypoxia A549 cells was significantly higher than that of normoxia cells.The green fluorescent marker of exosomes,PKH67,was distributed inside of the cell.Cell scratch test showed that the width of H-EXO group was much smaller than that of N-EXO group at 12,24 and 48 hours after exosomes treatment (t =2.96,6.76,3.35,P ＜ 0.05).Transwell assay showed that the number of transmembrane cells in the H-EXO group was more than that in the N-EXO group and the control group (t =4.84,7.88,P ＜ 0.O1).The expression levels of MMP2 (t =4.70,3.21,P＜0.05) and MMP9 (t =5.61,3.76,P＜0.05) in the supernatant of H-EXO group were significantly higher than those in the control and N-EXO groups.Cell survival assay showed that the D0 values of control,N-EXO and H-EXO group were 2.614,2.552 and 4.50 respectively,indicating that H-EXO could enhance radioresistance of A549 cells significantly.Conclusions This study finds that the number of exosomes released from A549 cells was increased under hypoxic condition but its size becomes smaller than that under normoxia.Hypoxic exosomes can promote the migration of normoxia cells andenhance cell radioresistance as well.
ABSTRACT
Objective To synthesize novel gold nanoparticles of GAL-PEG-GNPs,study its radiation effect on hepatocellular carcinoma cells HepG2 cells in vitro,and investigate the underlying mechanisms.Methods GAL-PEG-GNPs were synthesized and characterized successfully.HepG2 cells were divided into three groups of control,GNPs and GAL-PEG-GNPs.The cytotoxicities of these compounds were tested by the CCK-8 assay and their IC50 values of HepG2 cells were calculated.Cell uptake of nanoparticles was detected by TEM and ICP-MS.The radiosensitization effect of nanoparticles was tested by the colony formation assay.Cell cycle distribution was detected by FCM.The expressions of CAT,SOD,and total GSH were detected with a microplate reader,and the expressions of apoptosis-related proteins were tested by Western blot.Results The GNPs and GAL-PEG-GNPs had absorption peaks at 520 and 530 nm,respectively,and their diameters were (22.6-±2.12) and (32.0 ± 1.41) nm detected by ICP-MS.The GAL-PEG-GNPs and GNPs had similar cytotoxicity profiles (P ＞ 0.05),while GAL-PEG-GNPs could be more effectively uptaken by HepG2 cells than GNPs.The sensitive enhancement ratio (SER) of GNPs and GAL-PEG-GNPs to HepG2 cells were 1.46 和 1.95,respectively.The percentage of cells at phase of G2/M in HepG2 population treated with GNP was higher than that of untreated cells (t =14.20,P ＜0.05).The protein expressions of Cytochrome C,Bax,Caspase-3,and Caspase-9 were upregulated while Bcl-2 expression was down-regulated in the cells treated with GNPs/radiation or GAL-PEGGNPs/radiation.The expressions of CAT,SOD and total GSH in the GNP treated groups were significantly decreased compared with the control group(t =12.34,29.39,12.85,P ＜ 0.05).Conclusions GALPEG-GNPs has obvious radiosensitization effect on HepG2 cells,which is related to the induction of cell apoptosis and the generation of free radicals.
ABSTRACT
Objective To study the synergism effect of 131 I-Herceptin and high-energy X-ray on HER2 overexpressed breast cancer SK-BR-3 cells.Methods The protein expression and gene amplification of human epidermal growth factor receptor-2 ( HER2 ) in SK-BR-3 cells were identified by immunohistochemistry and fluorescence in situ hybridization ( FISH ) method, 131 I-Herceptin was prepared by iodogen method, and the IC15 concentration of 131 I-Herceptin on SK-BR-3 cell were selected by MTT method.The cells were divided into control group and drug group according to 131 I-Herceptin used or not, and were delivered five different doses of external irradiation (0,2,4 and 6Gy), and the synergism effect was detected by colonogenic assay.The cells were divided into blank group, drug group(131I-Herceptin), X-ray group(2 Gy external irradiation) and combination group (131I-Herceptin+2 Gy external irradiation), the apoptosis rate and death rate were detected by AO/EB method and cell cycle were detected by flow cytometry.Results The labling rate, radiochemical purity and specific radioactivity of 131 I-Herceptin were 86.8%, 93.9% and 868.3 μci/mg, respectively.The IC15 of 131 I-Herceptin was 15.625μci/mL.131 I-Herceptin and high-energy X-ray significantly reduced surviving fraction ( SF) ( F=628.888,F=964.97,P<0.05) and there were interactions between them (F=113.046,P<0.05).There were significant differences in apoptosis rate and death rate among blank group, drug group, X-ray group and combination group(F=103.324,F=13.33,all P<0.05),and there were significant differences of pairwise comparison (P<0.05).After irradiation and 131I-Herceptin administration, the cell cycle changed obviously from G1-phase to G2-and S-phase.Conclusion 131 I-Herceptin combined with high-energy X-ray has the synergism effect on HER2 overexpressed breast cancer SK-BR-3 cells.
ABSTRACT
Objective To explore the early diagnostic value of Golgi membrane protein 73 (GP73),alpha-fetoprotein (AFP) and alpha-fetoprotein-L3 (AFP-L3) in patients with high risk of primary hepatic carcinoma (PHC).Methods Sixty-four cases of PHC were selected as the PHC group,60 cases of liver cirrhosis(LC) as the LC group,53 cases of hepatitis as the hepatitis group and 51 healthy checked-up people as the control group.Enzyme-linked immunosorbent assay (ELISA) was used to detect the serum level of GP73 in all the cases.AFP-L3 was isolated by using affinity micro centrifugal column,AFP and AFP-L3 were detected with chemiluminescent immunoassay and then the proportion of AFP-L3 was calculated.Results The positive rate of serum GP73,AFP and AFP-L3 in PHC group was significantly higher than that in LC group and hepatitis group [78.1％ (50/64)vs.25.0％ (15/60),17.0％ (9/53);48.4％ (31/64) vs.31.7％ (19/60),22.6％(12/53) ;53.1％(34/64) vs.30.0％(18/60),20.8％(11/53)] (P ＜ 0.05),In control group,GP73,AFP,AFP-L3 was no positive.The levels of GP73,AFP and AFP-L3 in PHC group were significantly higher than those in LC group,hepatitis group and control group [(245.69 ± 89.18)μ g/L vs.(116.37 ±38.52),(97.29 ± 24.58),(23.48 ±9.12) μ g/L; (403.27 ± 128.46) μg/L vs.(75.62 ± 19.35),(66.49 ± 15.14),(3.46 ± 1.02) μg/L; (15.64 ±3.19)％ vs.(5.24 ± 1.15),(4.21 ± 0.96),(2.95 ±0.73)％] (P ＜0.05).The levels of GP73,AFP in LC group and hepatitis group were significantly higher than those in control group (P ＜ 0.05).The levels of GP73,AFP and AFP-L3 had no statistically significant difference between LC group and hepatitis group (P ＞ 0.05).Sensitivity and accuracy of three combined detection for PHC was 96.9％(62/64),91.7％(209/228),significantly higher than that of AFP,AFP-L3 single detection (P ＜ 0.05).GP73 single detection and any two combined detection was no significant difference in sensitivity and accuracy,compared with three combined detection (P ＞ 0.05).The levels of GP73 in PHC patients with different age,gender,serum level of AFP,TNM stage and tumor diameter had no statistically significant difference (P ＞ 0.05).The levels of GP73 in PHC patients with positive HBsAg,extrahepatic metastases and LC had significant difference (P ＜ 0.05).Conclusions The diagnosis value of GP73 is evidently higher than AFP and AFP-L3 for PHC,and combined determination is superior to single marker.Combined determination enhances the degree of precision in populations with high risk of PHC diagnosis.
ABSTRACT
Objective To study the radiosensitizing effect of nano-gold nano-silver particles in hepatocellular carcinoma cells (HepG2) in vitro and the possible mechanisms.Methods MTT assay and clonogenic assay were performed to determine the killing effect of nano-gold and nano-silver particles in HepG2 cells.Flow-cytometry was used to measure cell apoptosis and cell cycle distribution.Western blotting was used to measure the expression of Caspase-3,Bax and Bcl-2.ELIASA was used to determine the content of catalase (CAT),superoxide dismutase (SOD),and total glutathione (GSH).Results Nano-gold and nano-silver particles inhibited the proliferation of HepG2 cells with IC50 of 6.51 μg/ml and 2.47 μg/ml,respectively.Nano-gold and nano-silver particles significantly enhanced the radiosensitivity of HepG2 cells.Obtained by Dq,the SER of 1/5 IC50 nano-gold and nano-silver particles were 1.37 and 1.48,and 1/10IC50 with 1.11 and 1.09.Nano-gold and nano-silver particles increased the expression of Caspase-3 and Bax and reduced the exprcssion of Bcl-2.CAT,SOD and total GSH were significantly reduced.Conclusions Nano-gold and nano-silver particles can enhance the radiation sensitivity of HepG2 cells.Specific sensitizing mechanism may be the activation of the mitochondrial apoptosis pathway and the induction of reactive oxygen species in apoptotic pathways,which ultimately induces apoptosis.