AIM:To investigate the regulatory role of hypoxia mimic reagent cobalt chloride ( CoCl2 ) on cave-olin-1 (Cav-1) generation and the influence of Cav-1 on the abilities of migration and invasion of human lung adenocarcino-ma A549 cells.METHODS:The concentrations of Cav-1 and hypoxia-inducible factor ( HIF)-1αin pleural effusion of the patients with lung cancer ( MPE) or tuberculous pleurisy ( TBPE) were detected, and the correlation was also compared. A549 cells were treated with CoCl2 at different concentrations and time in the presence or absence of HIF-1αinhibitor YC-1.The concentrations of Cav-1 and HIF-1αin the cell supernatants were measured by ELISA.The effects of Cav-1 induced by CoCl2 on the migration and invasion of A549 cells were determined by scratch test and Transwell invasion trial, respec-tively.RESULTS:The levels of Cav-1 and HIF-1αin MPE were significantly higher than those in TBPE.There was a highly positive correlation between Cav-1 and HIF-1αlevels in the pleural effusion.CoCl2 induced the generation of Cav-1 and HIF-1αin A549 cells in a concentration-and time-dependent manner, the peak occurred at 200 μmol/L or 24 h, while the concentration over 200 μmol/L or after treated over 24 h, a concentration-or time-dependent inhibition was ob-served.HIF-1αinhibitor YC-1 concentration-dependently inhibited the generation of HIF-1αand Cav-1 induced by CoCl2 in A549 cells.CoCl2 enhanced A549 cells migration and invasion, with 200 μmol/L played the strongest role, which were down-regulated significantly in the presence of YC-1.CONCLUSION:The alteration of hypoxia-induced Cav-1 generation might be involved in the migration and invasion of A549 cells.A possible role for HIF-1αis indicated in Cav-1 generation.

Objective To investigate the effect of S-1 on serum thymidine kinase-1 ( TK1 ) , vascular endothelial growth factor ( VEGF ) and insulin-like growth factor-1 (IGF-1) and tumor biomarkers, estrgen levels and quality of life in patients with advanced breast cancer.Methods 69 cases with advanced breast cancer from January 2014 to May 2015 in the hospital were selected and randomly divided into two groups.34 cases in control group were treated by conventional therapy, and 34 cases in experimental group were treated with S-1.Serum TK1, VEGF and IGF-1, tumor biomarkers, estrogen level and quality of life score were compared pre-and post-treatment.ResuIts Compared with control group, the levels of VEGF, IGF-1 and TK1 were lower (P<0.05), serum CA125, CEA and CA15-3 concentrations were lower (P<0.05),the LH, E2 and E1 levels were lower (P<0.05) and the survival quality score was higher in experimental group ( P<0.05 ) .ConcIusion S-1 has better clinical curative effect in treatment of patients with advanced breast cancer, and effectively reduce serum TK1, VEGF, IGF-1, tumor biomarker levels, improve the quality of survival, which has important significance.

Objective To explore the expression of DKK-1 and its clinical value in patients with e-sophageal cancer. Methods The levels of serum DKK-1 were measured by sandwich enzyme linked immu-nosorbent assay(ELISA) in 80 patients with esophageal squamous cell cancer and in 35 healthy subjects. The sensitivities of serum DKK-1, CEA and CYFRA21-1 in the patients with esophageal squamous cell cancer were calculated and analyzed to compare their values in diagnosis of esophageal cancer. Results The serum DKK- 1 levels in patients with esophageal squamous cell cancer were significantly higher than those of healthy subjects(P<0.05). According to the highest value of medicine reference of DKK- 1 (14. 54 ng/ml), its sensitivity and specificity were 66. 25% and 82. 86% respectively; There was a significant difference between the sensitivity of DKK-1 in patients with esophageal squamons cell cancer(66. 25%) and that in healthy subjects(17.14%) (P<0.05). The sensitivitis of serum DKK- 1 in patients with esophageal squamous cell cancer were associated with tumor size, TNM stage and lymph node metastasis(P <0.05),but not associated with gender,age and differentiation grade(P>0.05). And the sensitivity of DKK-1 (66. 25%) was significantly higher than that of CEA, CYFRA21-1 (39. 44% ,25. 35%, P <0. 01). Conclusion DKK-1 played a very important role in the growth and metastasis of esophageal cancer and may be a new tumor marker or an important index in the diagno-sis of esophageal cancer.

AIM: To investigate the anti-cancer effect of cytotoxic T lymphocytes (CTL) activated by sensitized dendritic cells (DCs). METHODS: Immature DCs were induced in vitro from peripheral blood monocytic cells (PBMC) and sensitized by adding tumor cells antigen extract. DCs were identified by their morphology and surface markers. MTT assay was used to evaluate the killing activity of CTL activated by sensitized DCs. The effects of specific CTL cells on inhibiting transplanted tumor HT-29 growth and on preventing HT-29 tumor generation were evaluated by injecting CTL into nude mice. RESULTS: After cultured for seven days, a large number of activated DCs were obtained with typical morphology, extensive stimulatory proliferation capacity and high CD80 (63.5%), CD83 (67.6%) and CD3/HLA-DR (83.2%) expressions. The killing activity of CTL at 20∶1 ratio of effective cells to target cells was more than 75% to tumor cells, 35%-45% to homologous cell line and weaker to other germ cell line (P

AIM: To investigate the inhibitory effect of arsenic trioxide on human glioma cell line U251 growth, change of gene expression and intracellular calcium content. METHODS: MTT method was used to observe the growth inhibition effect. Cell cycle, positive rate of proliferation cell nuclear antigen (PCNA), apoptosis associated protein Fas and Bcl-2, and intracellular calcium ion (IECa~ 2+ ) levels were measured by flow cytometry in U251 cells treated with different doses of As_2O_3. Apoptosis was detected with annexin V-FITC+PI dual parameter. RESULTS: As_2O_3 inhibited the growth of U251 cells dramatically. There were obvious dosage-effect and time-effect correlations (P0.05). The cell cycle was arrested in G_2M phase. Apoptosis occurred in U251 cells treated with As_2O_3 by annexin V-FITC+PI dual parameter detection. CONCLUSION: As_2O_3 inhibits the growth of U251 cells in vitro dramatically and induces apoptosis. The mechanism is probably associated with the improvement of Fas expression and IECa~ 2+ levels, decrease in PCNA protein expression and cell cycle arrest.

Objective Studies show that the ERCC1 gene may be involved in secondary cisplatin resistance.This article aims to investigate the effects of shRNA targeting silencing excision repair cross-complementation group 1 (ERCC1-shRNA) on the proliferation and apoptosis of lung cancer A549/DDP cells treated with different concentrations of cisplatin.Methods Lung cancer A549/DDP cells were divided into a negative control, a blank control, an ERCC1-shRNA1, and an ERCC1-shRNA2 group.Human interfering RNA (RNAi) targeting the human ERCC1 gene was constructed and transfected into the A549/DDP cells using Lipofectamine 2000.The mRNA and protein expressions of ERCC1 in the A549/DDP cells were detected by real-time PCR and Western blot respectively, the proliferation-inhibition rate was assessed by MTT, and their cell cycle and apoptosis were determined by flow cytometry.Results ERCC1-shRNA was successfully constructed and transfected into the A549/DDP cells.Both the mRNA and protein expressions of ERCC1 were significantly lower in the ERCC1-shRNA1 (0.20±0.04 and 0.24±0.10) and ERCC1-shRNA2 (0.47±0.28 and 0.37±0.11) than in the negative control (0.96±0.12 and 1.32±0.13) and blank control groups (0.84±0.07 and 1.45±0.23) (P<0.01).Compared with the negative and blank control groups, the ERCC1-shRNA1 group showed a significantly decreased IC50 value (16.71±2.33 and 16.69±1.69 vs 7.78±0.54, P<0.01) and an increased proportion of G0/G1 phase cells ([72.87±3.23] and [71.75±4.56] vs [82.99±4.23]%, P<0.05), with the cell cycle arrested in the G0/G1 phase.The apoptosis rate of the cells in the ERCC1-shRNA1 group was remarkably lower after treated with cisplatin at the concentrations of 6.25 and 12.5 μg/mL than at 0 μg/mL ([8.17±0.65] and [11.91±1.41] vs [29.97±3.14]%, P<0.05).Conclusion ERCC1-shRNA can inhibit the proliferation and enhance the apoptosis of A549/DDP cells by silencing the expression of the ERCC1 gene.

Objective To explore the relationship of chemotherapy sensitivity and expression of multidrug resistance genes and apoptosis regulation genes in human breast cancer cell lines.Methods MTT assay was used to detect the sensitivity to adriamycin(ADM),cisplatinum(DDP),mitomycin C(MMC),fluorouracilum(5-Fu),carmustine(BCNU) in five breast cancer cell lines including Bcap37,MCF-7,T47D,MDA-MB-231and MDA-MB-435.Multidrug resistance genes including P-glycoprotein(P-GP),Glutsthione-s-transferases-?(GST-?),Lung resistance protein(LRP),multidrug resistance related protein(MRP),MGMT and apoptosis regulation genes FAS,BCL-2,P53 and P16 were examined by flow cytometry(FCM).Results The chemotherapy sensitivity was obviously divergent in different breast cancer cell lines.The correlation analysis showed that there were positive correlations between the sensitivity to 5-Fu in breast cancer cell lines and the expression of P16.There was no correlation among the sensitivity to other drugs and expression of other genes.Conclusions The sensitivity to 5-Fu is related to the expression of P16 in breast cancer cell lines.

Purpose:To study the relation between regional lymph node metastasis and nm23 H 1 protein expression and DNA ploidy of esophageal squamous cell carcinoma patients. Methods:Flow cytometry was used to analyze nm23 H 1 protein and DNA ploidy in surgical specimens from the patients with esophageal carcinoma. The object tissues included: the primary tumor tissue, the pericancerous mucosa, the incisional margin and the regional lymph node. Results:There were obvious differences between nm23 H 1 protein expression in lymph node metastasis positive and negative groups, primary tumor tissue and pericancerous mucosa, cancer tissue and the incisal margin( P 0.05).The differences of DI were distinct between cancer tissue and the pericancerous mucosa, cancer tissue and the incisal margin, cancer tissue and the lymph node( P 0.05).In corresponding tissues of lymph node metastasis positive and negative groups, nm23 H 1 protein expression was different( P 0.05). As to DI, it was different only among pathological grades( P 0.05). Conclusions:nm23 H 1 protein higher expression can produce certain inhibitory effect on metastasis of esophageal carcinoma. Combining DNA ploidy detection can help to understand the biological behavious and deduce the prognosis of esophageal carcinoma.

Purpose:To study COX 2 protein expression and its significance in familial adenomatous polyposis (FAP) and rectal cancer tissues. Methods: Flow cytometry was used to detect COX 2 protein expression in surgical specimens from 12 normal rectal mucosa tissues, 19 FAP tissues, 7 transformed FAP tissues and 36 rectal cancer tissues.Results:The COX 2 protein expression was negative in the 12 normal rectal mucosa tissues ( FI=0.67?0.28). The COX 2 protein expression was positive in 18 (94.74%) of the 19 FAP tissues (FI=1.97?0.36),the 7(100%) transformed FAP tissues(FI=2.02?0.24) and the 36(100%) rectal cancer tissues (FI=2.14?0.31). The COX 2 protein and positive expression rates showed that there were obvious differences between normal rectal mucosa tissues and FAP tissues, transformed FAP tissues, rectal cancer tissues( P 0.05). Conclusions:It is suggested that the detection of COX 2 protein expression can have important significance for predicting adenomatous polyposis carceration and chemo interfering carceration progress. [