Objective To investigate the role of Dicentrine in regulating lipopolysaccharide(LPS)-induced inflammatory responses in RAW264.7 macrophages and its underlying mechanisms.Methods RAW264.7 macropha-ges were cultured and pretreated with different concentrations of Dicentrine(250,500,1 000 μmol/L).An inflam-matory response model was established using LPS stimulation(model group),with a control group receiving no treatment.Cytotoxicity of Dicentrine was evaluated by MTT assay.Nitric oxide(NO)levels in cell culture supernatants were measured using the Griess method.Fluorescence microscopy was employed to observe in-tracellular reactive oxygen species(ROS)accumulation and apoptosis.Finally,mRNA expression levels of in-flammatory cytokines including IL-1β,IL-6,inducible nitric oxide synthase(iNOS),and tumor necrosis factor-α(TNF-α)were analyzed by qPCR.Results MTT assay results indicated that Dicentrine exhibited no cyto-toxic effects at concentrations below 1 000 μmol/L.Griess method and fluorescence microscopy showed signif-icantly elevated NO and ROS levels in the model group compared to the control group(P<0.05).Compared with the model group,the 250,500,and 1 000 μmol/L Dicentrine groups demonstrated significantly reduced NO and ROS levels(P<0.05).qPCR analysis revealed significantly increased mRNA expression of IL-1β,IL-6,iNOS,and TNF-α in the model group versus the control group(P<0.05).Compared with the model group,the 250,500,and 1 000 μmol/L Dicentrine groups showed significantly decreased mRNA expression of IL-1β,IL-6,iNOS,and TNF-α(P<0.05).Conclusion Dicentrine regulates macrophage apoptosis,thereby ef-fectively inhibiting LPS-induced inflammatory responses in macrophages.

Objective To explore the effect of Yes-associated protein 1 (YAP1) and potential mechanism on erlotinib ( ER) resistance in lung adenocarcinoma.Methods In PC-9 cells and acquired ER resistant PC-9 ( PC-9/ER) cells, the expression changes in YAP1 gene were measured by quantitative real-time PCR( RT-qPCR) and Western blot.Indirect immunofluorescence was adopted to observe the location of YAP1.PC-9/ER cells were treated with the verteporfin ( VP, YAP1 inhibitor) for 24 h and 48 h, respectively.Expression changes in mRNA and proteins of YAP1, AKT and p-AKT were detected in the presence or absence of VP.The effect of VP was analyzed by drug resistance index using Cell Counting Kit 8(CCK-8) assay.Results The resistance index of PC-9/ER cells was (99.80 ±25.81).Compared with PC-9 cells, the expression levels of YAP1 mRNA and protein were increased in PC-9/ER.The inhibitory efficiency of VP was (50.96 ±5.86)%, and the levels of AKT and p-AKT proteins were down-regulated by the inhibition of YAP1 simultaneously.The half maximal inhibitory concentration (IC50) of PC-9/ER decreased from (11.10 ±2.72) to (1.47 ± 0.32)μmol/L (P =0.024).Resistance index was reduced to one eighth of the original.Conclusion These results indicate that the YAP1 mediates ER resistance in lung adenocarcinoma.Suppression of YAP1 can reduce the resistance through PI3K/AKT signaling pathway.Therefore, YAP1 may be a potential target for lung cancer gene therapy.

Objective To investigate the value of detecting plasma microRNA‐125a‐3p(miRNA‐125a‐3p) ,IGF‐2 on monitoring invasion and metastasis in NSCLC ,and to study the correlation between miR‐125a‐3p and IGF‐2 .Methods miR‐125a‐3p transcripts of 20 controls ,73 NSCLC were performed in plasma by quantitative reverse transcription‐polymerase chain reaction(qRT‐PCR) and PCR data was analyzed by the 2‐ΔΔCT method .The expression of IGF‐2 in plasma was detected by ELISA .Results The expression of miR‐125a‐3p in stage Ⅲ /Ⅳ was lower than stage Ⅰ/Ⅱ and the controls(P=0 .001 ,P=0 .005) .There was no statistical differ‐ence between the stage Ⅰ /Ⅱ patients and the controls(P=0 .776) .The expression of miR‐125a‐3p was related with lymph node metastas ,lower expression in positive lymph node metastasis (P=0 .003) .The expression of IGF‐2 in stage Ⅰ /Ⅱ 、stage Ⅲ /Ⅳ was higher than the controls(P=0 .036 ,P=0 .011) .There was no statistical difference between the stageⅠ/Ⅱ and stage Ⅲ/Ⅳ (P=0 .451) . The expression of IGF‐2 was related with lymph node metastas ,higher expression in positive lymph node metastasis (P=0 .037) .The re‐sults showed a negative correlation between miR‐125a‐3p expression and IGF‐2 in plasma(r= -0 .280 ,P=0 .007) .Conclusion Low ex‐pression of miR‐125a‐3p and high expression of IGF‐2 in plasma may play a role in invasion and metastasis of NSCLC .miR‐125a‐3p may play a negative regulatory role on IGF‐2 .

BACKGROUNDAs a new member of inhibitor of apoptosis protein(IAP) family,Livin,especially Livin α,is known to be involved in occurrence and development of lung cancer.Livin is an important mechanism of chemotherapy resistance of lung cancer cell.The aim of this study is to set up Livin isoform(α & β)-specific gene silencing system in SPC-A1 cells by gene transfection and RNA interference(RNAi),and to explore the different functions and value of the isoforms in enhancing chemosensitivity of SPC-A1 cells.

METHODSLivinα+β,Livinα and Livinβ specific siRNA were expressed stably in SPC-A1 cells,respectively.MTT was performed to study sensitivity of the cells to chemotherapy drugs.In vivo experiment was performed to test sensitivity of mouse bearing tumor to cisplatin after gene silencing of Livin.

RESULTSAfter silencing of Livinα+β,Livinα and Livinβ genes,sensitivity of SPC-A1 cells to many chemotherapy drugs(including cisplatin,carboplatin,cyclophosphamide and adriblastine) was markedly increased(P < 0.05).Among them,gene silencing of Livinα+β showed the strongest enhancement effect on chemosensitivity of SPC-A1 cells(P < 0.01).Animal experiment showed that tumor inhibition rate of pSilencer-Livinα+β,pSilencer-Livinα and pSilencer-Livinβ groups was 146.1%,130.7% and 110.5%,respectively.

CONCLUSIONSThe results suggest that Livin isoform,especially Livinα+β is hopeful to be a molecular target for increasing sensitivity of lung cancer cell to chemotherapy.Gene silencing may be a new means of gene therapy for non-small cell lung cancer.

BACKGROUNDGensing Rg3 is an active component from ginseng. The aim of this study is to observe the clinical anticancer effect of Rg3 in combination with chemotherapy regimen NP (vinorelbine+cisplatin) in advanced non-small cell lung cancer (NSCLC).

METHODSStage III-IV NSCLC patients confirmed by pathology or cytology all received vinorelbine plus cisplatin for at least two cycles, and were randomized into two groups: patients in arm A also received placebo twice a day, while patients in arm B received two tablets of Rg3 twice a day for at least two months. The endpoints of the study were the efficacy, survival and tolerance of patients.

RESULTSFrom July 2000 to May 2002, 115 patients were enrolled into the trial. The patients' characteristics were well balanced in the two groups. Sex of patients: male, 79; female 36. Types of pathology: adenocarcinoma, 71; squamous cell carcinoma, 29; adenosquamous carcinoma, 8; others, 7. TNM stage: stage III, 45; stage IV, 70. Prior chemotherapy: with, 17; without, 98. Prior radiotherapy: with, 15; without, 100. Prior surgical treatment: with, 23; without, 92. Nine patients discontinued from the trial due to severe adverse effects (5) and other reasons (4), so there were 106 patients evaluable for clinical efficacy. The response rate was 14.5% (8/55) in arm A, and 33.3% (17/51) in arm B (P=0.011). The survival time in arm A was 9.7 months (mean) and 8.0 months (median), and 15.3 months (mean) and 10.0 months (median) in arm B (P=0.0088).

CONCLUSIONSPreliminary results show improvements in response rate and survival time (median and mean) in Rg3 arm compared with placebo arm. It is worthy to confirm the results in further clinical trials.

Objective To cultivate,identify and sort bronchoalveolar stem cells(BASC)derived from normal adult mouse lung.Methods After enzymatic digestion of lung tissue with dispase and collagenase in combination,the Sca-1+ cells were isolated from the obtained pulmonary cells by magnetic cell sorting.These Sca-1+ cells were cultured in dishes coated with collagen and mouse fibroblast cell line Swiss-3T3 under a serum-free culture system for BASC,which were identified by the dual-color immunofluorescent staining clara cell specific antigen(CCA)and surfactant protein C(SP-C).Finally,these pure BASC were isolated by the flow cytometry.Results One lung of normal adult mouse could yield(1.6-1.8)?107 nucleated cells in this enzyme digestion procedure.The percentage of Sca-1+ cells we sorted from lung tissue was much higher than the unsorted [(87.3?5.9)% and(9.6?1.8)%,P

Purpose:To explore different functions and values of Livin ? and Livin ?-specific silencing in inducing SPC-A1 cell apoptosis, respectively. Methods:First, Livin ?+?, Livin?and Livin ?-specific siRNA were steadily expressed in SPC-A1 respectively. Next, after isoform-specific gene silencing, biological changes with regard to cell morphology and proliferation of lung cancer cell were observed. Then, TUNEL and FCAS were performed to test the rate of apoptosis. Lastly, statistical analysis was performed to explore the different functions and values of isoform-specific gene silencing in inducing lung cancer cell apoptosis.Results:After gene silencing of Livin ?+?, Livin ? and Livin ?,there was an evident decrease in colony forming ability of SPC-A1 compared with the control(P

Objective To express miR-122 in HeLa cells. Methods pAVU6+27-mir122, an eukaryotic expression vector of miR-122, was transfected into HeLa cell line by electroporation. G418-resistant clones were screened for cell clones with stable transfection. Northern blotting and dot blotting were used to detect miR-122 expression in the transfected HeLa cells. Results Northern blot and dot blot analyses demonstrate miR-122 expression in both transfected HeLa cells and murine hepatocytes, but miR-122 expression was not found in HeLa cells transfected with pAVU6+27. Conclusion The expression of miR-122 in HeLa cells may provide the basis for further studies of its targets and biological functions.