OBJECTIVE: To study the effect of vinorelbine tartrate lipid microsphere(NVB-lip) injection on growth of human breast cancer cells in nude mice. METHODS:35 female BALB/c-nu mice were inoculated with human breast cancer BCAP-37 cells. These mice were divided randomly into 5 groups for further treatments:NVB-lip high dose group(NVB-lip 10 mg/kg),medium dose group(NVB-lip 5 mg/kg),low dose group(NVB-lip 2.5 mg/kg),positive drug control group(NVB-free 5 mg/kg) and negative control group(only lipid microsphere). There were 7 mice each group. Injection with NVB-lip was done after 6-8 days when the tumors shaped. NVB-lip was injected intravenously every 3 or 4 days each week for 6 times in total. Weights and tumor volumes(V T) of mice were observed and measured on No. 4,7,11,14,18 and 23 days from the first injection. Relative tumor volumn(V RT),relative tumor proliferation rate and tumor inhibition rate were calculated. RESULTS:The weights of nude mice for different groups increased slowly but the differences were not statistically significant. Compared with the negative group,the V RT and relative tumor proliferation rates of NVB-lip groups were reduced but tumor inhibition rate was increased. The differences were statistically significant(P0.01). CONCLUSION:The inhibition of NVB-lip high dose group(2.5-10 mg/kg) was more obviously than that of the other groups. NVB-lip may h ave c linical a pplications.
OBJECTIVE: To understand the relationship between aberrant expression of E-cadherin in nasopharyngeal carcinoma(NPC) and clinicopathological features of NPC patients for clinical applications. METHODS: International publications about E-cadherin and NPC were selected from the Cochrane Library, Pubmed,EMbase database,and Chinese publications were from CBM,CNKI. All analyses were performed by software STATA 11.0. Abnormal expression,degree of differentiation,T stage,lymph node metastasis,and clinical stage were analyzed using pooled odds ratio(OR) with 95% confidence interval(CI). RESULTS:A total of 16 studies including 1040 patients were enrolled in the Meta-analysis. It was shown that expression of E-cadherin was significantly increased in NPC compared to that in normal tissues(OR=0.096,95% CI:0.036-0.252,P=0.000). In addition,the aberrant expression of E-cadherin was significantly associated with the degree of differentiation,lymph node metastasis and clinical stages(OR= 0.232,95% CI:0.061-0.884,P=0.032;OR=0.324,95% CI:0.198-0.53,P=0.000;OR= 0.441,95% CI: 0.232-0.835,P=0.012). CONCLUSION:NPC often showed aberrant expression of E-cadherin. In addition,patients with the aberrant expression were prone to having tumors at lower differentiation,with lymph node metastasis and at late clinical stage. E-cadherin may be an indicator for differentiation,lymph node metastasis and clinical stage.
OBJECTIVE: To observe the photobleaching characters of sinoporphyrin sodium(DVDMS) in different solutions and skins of animal. METHODS:Absorption peaks of DVDMS(10,20,40 μg/m L) in normal saline(NS),RPMI-1640 medium(RPMI-1640),RPMI-1640 medium containing 10% fetal calf serum(RPMI-FBS) and suspension of Ha Ca T cells were measured using micro-spectrophotometer. Using DVDMS solutions with concentrations of 1,2.5,5,10,20 and 40 μg/m L in the 4 solvents above,photobleaching of DVDMS after laser radiation for 3,6,10,20,40,50,60,120,180 min was observed using microplate reader. In vivo image acquisition system was used to study the distribution of DVDMS in animal skins,and the bleaching after irradiation in psoriasis-like animal skins were also observed. RESULTS:DVDMS with concentrations 10,20 and 40 μg/m L showed obvious characteristic absorption peaks in the 4 solutions. Photobleaching occurred in the 4 solutions under the irradiation conditions used in previous pharmacodynamics studies(laser radiation was 3 minutes). Two-phase processes photobleaching(fast first phase and second slow phase) of DVDMS were observed in 180 min. The distributions of DVDMS in skins of normal and psoriasis-like animal were uniform,obvious photobleaching occurred in skins of psoriasis-like animal after laser 2irradiation with dose of 9.6 J/cm. CONCLUSION:In vitro photobleaching of DVDMS was influenced by light dose,initial concentrations and solvents. Secondary-dynamic model was suitable to describe the photobleaching progress. Photobleaching occurred in skins of psoriasis-like animal after laser irradiation.
OBJECTIVE: To understand autophagy induction by tert-butyl hydroperoxide(t-BHP) in human hepatic cell line L02 and to explore the involvement of mitochondria. METHODS:L02 cells were treated with different concentrations of t-BHP(100,200,400,800,1 000 μmol/L) with or without specific inhibitors(mitochondria targeted antioxidant Mito Q or p38/MAPK inhibitor SB203580). Autophagy markers LC3-II and p62 as well as the p38/MAPK pathway proteins were detected by immunofluorescence and/or Western blot. Mitochondrial reactive oxygen species(ROS) were measured by flow cytometry with Mito-SOX Red kit. RESULTS:The high concentrations of t-BHP(≥800 μmol/L) induced the accumulation of LC3 fluorescence puncta,with LC3-II protein level increased and p62 protein level decreased significantly. These changes accompanied the elevated mitochondrial ROS and the activated p38/MAPK pathway. Pretreatment with mitochondria targeted antioxidant Mito Q or specific p38/MAPK inhibitor SB203580 attenuated these changes which were induced by t-BHP(1 000 μmol/L). CONCLUSIONS:We established the autophagy model induced by t-BHP in human hepatic cell line L02. Mitochondrial ROS and p38/MAPK pathway played critical roles in this process.
OBJECTIVE:To establish the early warning of the contamination of nano-SiO_2 particle,swimming behavior changes of zebrafish under nano-SiO_2 particle exposure were studied.METHODS:Based on behavior response of zebrafish,effects of nano-SiO_2 particles,including individual endpoints(swimming speed,swimming depth,turning frequency and acceleration) and group endpoints(distance and dispersion),were investigated by using Online Bio monitoring.RESULTS:Swimming behavior changes of zebrafish were affected significantly in 600 and 1 200 mg/L exposure dose groups.Swimming speed was decreased 30%-50%6 h after exposure.Changes of swimming depth lagged behind changes of swimming speed,which were decreased by more than 90%24 h after exposure.Group behavior of zebrafish was also changed,distance and dispersion were significantly decreased(all P0.01).CONCLUSION:After exposure,swimming behavior of zebrafish(swimming speed,swimming depth,turning frequency,distance and dispersion) were affected significantly with the increase of nano-SiO_2 particles concentration,which is consistend with the Stepwise Stress Model.
OBJECTIVE:Trichloroethylene (TCE),a common industrial contaminant in theenvironment,could induce hepatocellular carcinoma but not kidney cancer in mice. This study aimed to understand the molecular mechanisms of TCE hepatocarcinogenicity by examining the effect of TCE on proliferation-related mRNA expression and DNA methylation in mouse liver and kidney.METHODS:B6C3F1 male mice aged six weeks were randomly divided into 3 groups,with 4 mice in each group. Mice were fed with TCE at 0,500 and 1 000 mg/kg for 5 days. Using qPCR,we examined the mRNA expression of genes involved in cell proliferation and in the regulation of DNA methylation in TCE-exposed mouse liver and kidney. The promoter methylation status ofCdkn1a and the DNA methylation level of repetitive sequences were detected by combined bisulfite restriction analysis. RESULTS:Compared with control,TCE increased the mRNA expression of Cdkn1a,Jun andMki67 in a dose-dependent manner in mouse liver. Moreover,in mouse treated with TCE at 1 000 mg/kg,the mRNA expression of a number of key DNA methylation regulation genes includingDnmt3a,Dnmt3b andTet2 were down-regulated,while the mRNA expression level ofUhrf1 was up-regulated(allP<0.05). TCE also induced hypomethylation o f t he p romoter r egion o fCdkn1a i n m ouse h epatocytes. H owever,TCE should n o d etectable e ffects o n m RNA expression and DNA methylation in mouse kidney.CONCLUSION:Alterations in the mRNA expression of proliferation-related g enes a nd D NA m ethylation c hanges m ight b e c rucial t o T CE-induced h epatocarcinogenesis i n m ice.
OBJECTIVE: To evaluate the acute toxicity and mutagenicity of potassium 2-(1-hydroxypentyl)-benzoate(dl-PHPB). METHODS:KM mice were intravenously treated with dl-PHPB at the doses of 256.0、286.3、320.0、357.8、400.0 and 447.2 mg/kg. We observed the clinical signs of toxicity and calculated the median lethal dose(LD5 0) and LD50 95% confidence interval. Mutagenicity of dl-PHPB was studied by Ames test,bone marrow micronucleus study in male mice and in vitro CHL cell mammalian chromosome aberration assay. In Ames test,the tester strains used were Salmonella typhimurium TA97,TA98,TA100 and TA102,mutagenicity was evaluated with the plate incorporation method beginning at 0.5,5,50,500 and 5 000 μg per plate in the absence or presence of S9 metabolic activation. In bone marrow micronucleus study,KM male mice were intravenously treated with dl-PHPB at the dosse of 210、70 and 23.3 mg/kg. Bone marrow cells were harvested from mice at 24 hours after dosing. 1 000 PCEs per animal were examined microscopically for the presence of micronucleated polychromatic erythrocytes(MNPCEs). The in vitro CHL cell mammalian chromosome aberration study on dl-PHPB consisted of the premilinary toxicity assay and chromosome aberration assay,with and without S9 metabolic activation system, and chromosomal aberration percentage per dosewere analyzed. RESULTS:In the acute toxicity study,all mice after dosing developed transient toxic symptoms such as tachypnea,sautonomic activity decreased,prostration,then the dying state were observed at dosage of ≥ 320.0 mg/kg,mice appeared respiratory depression,cyanosis,loss of righting reflex,and died during 2 to 30 minute after administration. The mortality rate of mice treated with dl-PHPB from 256.0 to 447.2 mg/kg were 0,0,10%,30%, 70%,100%,respectively. The survival mice returned to normal after 30 min. In the 14-days observation period,there were no abnormal clinical signs in the surviving animals and the general autopsy showed no abnormality in major organs. The intravenous median lethal dose of dl-PHPB was 373.3 mg/kg in mice,LD50 95% confidence interval was 355.6 to 392.0 mg/kg. In the Ames assay,no positive mutagenic response was observed. In bone marrow micronucleus study,a single bolus intravenous administration of dl-PHPB at dosages of 23.3,70 and 210 mg/kg didnu0027t induce a significant increase in the number of MNPCEs in the bone marrow,MNPCEs were less than 4‰ at three dosages. In CHL chromosome aberration assay,dl-PHPB achieved a chromosome aberration of less than 5% in the presence and absence of S9 mixture. CONCLUSION:The intravenous median lethal dose of dl-PHPB was 373.3 mg/kg in mice,and dlPHPB was not mutagenic in these 3 screening mutation a ssays.
OBJECTIVE: To explore whether the DNA copy number changes in 5 gene fragments are specific abnormalities for bladder cancer in Chinese patients. METHODS:Real-time PCR was conducted to gauge DNA copy number alteration of the 5 gene fragments among commercial genome DNA of peripheral blood leucocyte(PBL) derived from Caucasian,genome DNA of PBL from Chinese healthy volunteers,the bladder cancer patients and their corresponding tumor tissues. RESULTS:Comparing with the Caucasian PBL,all the gene fragments revealed copy number changes with decreased CEP63,increased FOSL2,GHR and PAQR6,in the PBL of Chinese healthy volunteers,except ZFAND3. Comparing with the PBL of Chinese healthy volunteers,in the bladder tumor tissue samples,the copy number of CEP63,GHR and PAQR6 increased(all P0.05);whereas FOSL2(P=0.41) and ZFAND32(P=0.062) revealed no statistical difference(all P0.05). Comparing with the PBL of Chinese healthy volunteers,the copy number of CEP63 and PAQR6 increased but FOSL2 and ZFAND3 decreased in the PBL of bladder cancer patients(all P0.01). Comparing with the PBL of the bladder cancer patients,gain of copy number was observed with CEP63,FOSL2,GHR and ZFAND3(all P0.01),while PAQR6(P=0.325) exhibited no statistical difference,in the corresponding tumor tissues. CONCLUSION:Copy number variation of CEP63,FOSL2,GHR and PAQR6 was found between Chinese(Han) and Caucasian. Copy number gain of CEP63 and GHR may be involved in carcinogenesis of bladder cancer;while gain of PAQR6 was possibly associated with the genetic s usceptibility for this malignancy.
OBJECTIVE:Polyvinyl chloride(PVC) was used as matrix to prepare the positive reference material by adding dibutyl tin maleate.METHODS:Identification and impurity inspection were carried out for the matrix.The positive reference material,PVC film containing organic tin,was prepared by tape casting process.Sample extract was prepared for L929 cell culture medium.The optimal organic tin concentration was determined by cell morphology examination and MTT test.Homogeneity and stability test were performed for the positive reference material.RESULTS:Extract of PVC film containing 0.005%organic tin exhibited significant cytotoxicity in a dose-effect manner.The homogeneity test showed the statisticd F was 1.07 which was less than the threshold value F.PVC film containing0.005%organic tin exhibited stable positive reactions.Whilst the materials before and after aging test revealed that the tin contentobtained from extraction was uniform(P0.05).CONCLUSION:Extract of PVC film containing 0.005%organic tin showed appropriate cytotoxicity,PVC film containing organic tin had a good uniformity and stability,so it could suitable as a cytotoxic reference material.
OBJECTIVE: To study the effects of carboplatin combined with TRAIL(TNF- related apoptosis inducing ligand) on proliferation and apoptosis of human lung cancer A549 cells. METHODS:After treated by TRAIL,carboplatin or combination of two drugs,the proliferation of A549 was measured by MTS and apoptosis rate was evaluated by flow cytometry. The morphology was investigated by microscopy. RT-PCR and Western blot were used to examine the gene and protein expressions of DR4,DR5,Survivin and XIAP. RESULTS:Carboplatin or TRAIL alone or combined inhibited the proliferation of A549 cells in concentration-dependent(20-80 μg/mL) manner. Carboplatin combined with TRAIL exhibited a stronger inhibitory effect and induced apoptosis than when used alone. Carboplatin could down-regulate the protein expression level of Survivin and XIAP and increase the DR5 protein expression level,but have no effect on DR4 expression. CONCLUSION:Combination of carboplatin and TRAIL could inhibit proliferation and apoptosis via influencing the expressions o f Survivin,XIAP and DR5 in A549 c ells.
OBJECTIVE:To study the genotoxicity and apoptosis of the mice L5178 Y lymphoma cells which was induced by disinfection by-product dibromo acetonitrile in drinking water.METHODS:Cytokinesis-block micronucleus cytome assay(CBMN-cyt) and flow cytometry were used to evaluate genotoxicity and apoptosis in mice L5178 Y lymphoma cells treated with different concentrations of dibromo acetonitrile(0.1,1,5 and 10 mol/L).RESULTS:Compared with the negative control group,frequency of micronucleus(MN) of L5178 Y lymphoma cells in 1 and 5 μmol/L groups increased significantly(P0.05).The nucleoplasmic bridges(NPBs) in the 1 and 10 μmol/L treatment groups and the frequency of nuclear buds(NBUDs) in the 10 μmol/L treatment group all increased significantly(P0.05).However the nuclear divided index(NDI) in the 5 and 10 μmol/L treatment groups all decreased significantly(P0.05).Statistically significant increase of L5178 Y lymphoma cell apoptosis were observed in all dibromo acetonitrile treatment groups in comparison to the negative control group(P0.05).CONCLUSION:Disinfection by-product dibromo acetonitrile in drinking water could obviously induce genotoxicity and apoptosis in L5178 Y lymphoma cells.
OBJECTIVE: Try to identify single nucleotide polymorphism(SNPs),copy number variation(CNVs) and m RNA expression from the genetic or tumor markers that are associated with lymph node metastases in colorectal cancer. METHODS:Targeted next-generation sequencing(Illumina Hiseq) was applied to capture SNPs and CNVs in tumor-related candidate genes of tumor tissues and paired normal tissues adjacent to the tumors from 25 colorectal cancer specimens;real time-PCR was used to detect specific m RNA expression of tumor-related candidate genes(VEGFC, CCNA2,IL2,ABCG2,EGF and NFKB1) on chromosome 4 of 39 colorectal cancer patient specimens. RESULTS: The SNPs in SLC28A3,BRCA1,RRM2,PMS2,CDA,EPHX1,RALY,CD33,BCL10,ETV1, MST1 R, KMT2 B,BCL2,LSM3,TTF1,MAP3K1 genes were significantly correlated with lymphatic metastasis risk(P0.05). There were no significant differences of the CNVs in the DDR1, CYP21A2, SULT1A1,XRCC2,POU5F1,FLT1 genes between the positive lymph node metastasis group and the negative lymph node metastasis group(P0.05). Compared with non lymph node metastasis group,m RNA expressions of EGF and NFKB1 were both down-regulated in the lymph node metastasis group,and the difference was statistically significant(P0.05). CONCLUSION:SNPs in SLC28A3, BRCA1,RRM2,PMS2,CDA,EPHX1,RALY,CD33, BCL10,ETV1,MST1 R,KMT2B,BCL2,LSM3, TTF1,MAP3K1 genes and down-regulated expression of EGF and NFKB1 might be the potential genetic markers in lymph node metastasis of colorectal cancers. No CNV was found to be associated with lymph node metastasis in colorectal cancer in our study.
OBJECTIVE: To establish a high-throughput screening method of cell transformation assay for carcinogens detection using Bhas 42 cell line,and evaluate the potential carcinogenic risk of genistein. METHODS: We established Bhas 42 cell transformation assay and compared the effects of traditional foci formation as well as H2O2 treatment methods on data analysis. In the traditional foci formation method,cells were fixed and dyed on day 21 after plating,whereas in the H2O2 treatment method,cells were treated with H2O2 for 24 h before dyeing and the transformation ratio measured at D(450) on day 20. Then the transformation potential of genistein on Bhas 42 cells was determined by the H2O2 treatment method,after a dose range-finding assay was performed to estimate the concentrations of genistein to be used based on the growth rates of cells. RESULTS:Apparent increase in the number of wells with transformed foci were noted in the positive control(3-MCA) in the initiation assay and the positive control(TPA) in the promotion assay,compared with the negative control in the traditional foci formation method(P0.01). Meanwhile,apparent increase in the number of wells with transformed foci and D(450) values were noted in the positive control(3-MCA) in the initiation assay and the positive control(TPA) in the promotion assay,compared with the negative control in the H2O2 treatment method(P0.01). The effects of genistein on Bhas 42 cell line was also examined. Although the D(450) values showed no difference between negative control and groups treated with genistein(0.03,0.1,0.3,1,3 μg/m L) in the initiation assay,a significant difference was noted in the promotion assay with the concentrations of genistein ranging from 0.03 to 3μg/m L(P0.01). CONCLUSION:This study has clearly demonstrated the H2O2 treatment method as a reliable improvement for Bhas 42 cell transformation assay. This cell transformation assay combined with initiation and promotion models is an effective and practical screening platform for non-genotoxiccarcinogens.,and it will be extremely helpful in the early stage of new drug development. Genistein could be a potential non-genotoxic carcinogen,however,this result needs to be further confirmed.
OBJECTIVE: In this study,the toxicokinetics and toxicodynamics of chlorpyifos were assessed in rats following one single subcutaneous exposure. METHODS:Adult female SD rats,randomly divided into three dose groups(69.75,139.5,279 mg/kg) and one control group(0 mg/kg),received one subcutaneous injection. Blood and cerebral cortex samples were collected on 3,6,12,24,48 and 72 hours in each group,and 0-24,0-48,0-72 h urine samples were collected continuously. The indicators included chlorpyrifos(CPF) in serum,3,5,6-trichloro-2-pyridyol(TCP) in urine and serum,and acetylcholinesterase(ACh E) in serum and cortex. RESULTS:With the increase in the exposure dose,the concentration of serum CPF,serum TCP,urine TCP and inhibition of serum and cortex ACh E increased. The peak concentration of serum CPF and TCP were 6 and 12-24 h,respectively. Maximum inhibition of serum and cortex AChE occured at 24 and 24-48 h,respectively. Half-life(t 1/2) of serum CPF was 29-48 h,and 0-72 h area under the curve(AUC) was 31-110 h·μmol/L in each group. t 1/2 of serum TCP was 28-54 h,and 0-72 h AUC value was 897-3 450 h·μmol/L. Serum and cortex AChE activity demonstrated a positive correlation(P0.01),and serum AChE activity had higher inhibition(P0.01). There was a dose-response relationship between serum CPF and AChE,fitting by Dose Resp curve. CONCLUSION:External exposure dose of CPF was correlated with CPF,TCP and AChE activity. There was a dose-effect relationship between CPF internal level and serum AChE activity. Excretion of TCP in urine can be used as a biomarker of exposure. Serum AChE activity was an effective biomarker of cortex AChE activity.
OBJECTIVE: Build and validate PBTK/TD with growth function of the Juvenile rat following the oral exposure to chlorpyrifos(CPF). METHODS:Model construction and validation:to determine the model structure and parameters;establish the growth function of body weight for rats,and write the differential equations as well as the program. Using experimental data to optimize the model parameters,make Sensitivity Analysis and model validation. Animal experiment:21 days-old female SD rats were randomly divided into 1 control group and 3 treated groups(12.5,25.0 and 50.0 mg/kg) according to the weight. Serum and cerebral cortex were collected after single gavage at 1h,3 h,6 h,12 h and 24 h,then to determine the concentration of CPF and 3,5,6-trichloropyridinol(TCP) in serum,the activity of acetylcholinesterase(ACh E) in cerebral cortex. RESULTS:Experimental data showed that the concentrations of serum CPF and TCP increased to the peak level at the 3rd hour,then the concentrations decreased. With time,the activity of ACh E in serum and cerebral cortex decreased first and then rose slowly. The minimum activity of serum ACh E and cortex ACh E was observed at 6th hour and 12 th hour,respectively. The model predictions is consistent with the experimental results,while there were an overvalued or undervalued problem when using the model to make extrapolation. CONCLUSION:The model has reasonable structure and parameters,and can be used for extrapolation,especially in prediction of the individual average level,but the model still need further improvement.
OBJECTIVE: To study the inhibitory effect of sodium norcantharidin lipid microsphere injection on nude mice bearing human carcinoma A549. METHODS:Animal model of human lung cancer was established by inoculating human carcinoma A549 cells to BALB/c nude mice. 11 days after inoculation,mice were randomly divided into control group,positive control group,and other three groups with different doses of sodium norcantharidin lipid microsphere injection. The mice received physiological saline,2.5 mg/kg sodium norcantharidin injection,and 1.25,2.5 or 5 mg/kg sodium norcantharidin lipid microsphere injection at 1 d,4 d,8 d,11 d,15 d,18 d,and 22 d after grouping. The tumor volume(Vt),relative volume(VR T),and the increment rate(T/C) were measured at 7 d,14 d,and 24 d after the first injection. The weight of tumors and the growth inhibition rate in vivo were measured at 24 d after the first injection. RESULTS:The tumor volume and the relative volume of all treated groups were much smaller than the group with negative control 7 days after the first injection,and the difference was significant(P0.05). These effects showed drug dose- effect relationships(r7 d =0.994,r1 4 d =0.993,r 24 d =0.996). The tumor increment rate of all treated groups were lower than 25%,especially the medium dose and the high dose groups which dropped to below 20%. The average tumor weights of the treated groups were lower than the control group(P0.01). The growth inhibition rates were more than 50%. CONCLUSION:Sodium norcantharidin lipid microsphere injection effectively inhibited the growth and reduced increment rate of human carcinoma A549 transplanted in nude mice.
OBJECTIVE: To investigate the expression of silent information regulator 1(SIRT1) in bladder urothelial cell carcinoma(BUC),and analyze its clinical significance. METHODS:The expression of SIRT1 protein was assessed by immunohistochemistry in bladder sections of 19 normal bladders and 72 patients with BUC. We analyzed the relationship of SIRT1 and the pathological grade and clinical stage of BUC. RUSULTS:SIRT1 was observed in normal bladder samples and BUC.Normal bladder showed negative or weakly positive expression of SIRT1(31.58%),however BUC samples showed moderately or strongly positive expression(77.78%). Furthermore,the expression level of SIRT1 increased with tumor pathological grade and clinical stage. SIRT1 positive expression rates showed significant difference between various pathology grades(P0.05). CONCLUSION:SIRT1 was high expressed in BUC,and was related to its pathological grade and clinical stage,making it a useful tumor marker and a possible indicator for estimating the invasiveness and recurrence of BUC.Blocking SIRT1 expression and its functions may become targeted therapy of bladder cancer treatment.
OBJECTIVE:To study the effects of norcantharidin (NCTD) on human lung cancer cells,and investigate the mechanisms.METHODS:The growth inhibition of A549 cells treated with 0-240μmol/L NCTD for 0-72 hours was analyzed by MTT assay. The recovery growth and proliferation of A549 cells treated with 0-120μmol/L NCTD for 24 h was evaluated by MTT assay. The morphological changes of A549 cells treated with 40,50 and 60μmol/L NCTD for 0-72 h were examined under inverted microscope. The apoptosis and cell cycle changes of A549 cells treated with 40-60μmol/L NCTD were detected by flow cytometry.RESULTS:NCTD inhibited the growth of A549 cells in 30-240 μmol/L(P<0.05). Cell growth inhibition could hardly be detected 24 h after 30-240μmol/L NCTD was removed from culture medium. The vitality of A549 cells was fully recovered in 5 d after 30-60μmol/L NCTD was removed from culture medium. A549 cells showed G2-M phase block but no obvious apoptosis compared with the control group when treated with 40,50 and 60μmol/L NCTD for 0-72 h. CONCLUSION:NCTD (40-60μmol/L) inhibited human lung cancer A549 cell growth mainly by cell G2-M phase block.
OBJECTIVE:This study aimed to investigate the expression of integrinαvβ6 and JunB in oral squamous cell carcinoma (OSCC) and the correlation with clinicopathologic characteristics and prognosis.METHODS:The expression of i ntegrinα vβ6 a nd J unB p rotein w as d etected w ith i mmunohistochemical s taining a ssay i n 1 20 c ases o f O SCC a nd 1 5 samples of normal oral mucosa. The correlation with clinicopathologic characteristics and prognosis of the patients were evaluated. RESULTS:Integrinα vβ6 w as d etected i n b oth c ytoplasm a nd c ellular m embrane o f O SCC,but J unB p rotein w as l ocalized i n the nuclei of both OSCC and normal oral mucous tissues. The expressions of integrinαvβ6 and JunB protein in OSCC were significantly higher than that in normal oral mucosa (P<0.01). The expression of integrinαvβ6 was positively related to depth of invasion(T stage),tumor differentiation degree and lymph node metastasis(N stage) (P<0.05). The expression of JunB was positively related to depth of invasion and lymph node metastasis (P<0.05). Survival as a single factor analysis revealed that patients w ith o ver-expression o f i ntegrinα vβ6 h ad a s horter o verall s urvival(OC) (P=0.021). T hrough C ox m ultivariate r egression analysis,lymph node metastasis (N stage) was an independent risk factor that affected the prognosis. CONCLUSION: The over-expressions of integrinαvβ6 and JunB were common in OSCC and closely correlated with tumor invasion and metastasis. I ntegrinα vβ6 m ay b e u sed a s a p rognostic m arker f or t herapy.
