The differences in intracellular and extracellular protein expressions between human prostate cancer lines LNCap and DU145 were examined. The proteins of the two cell lines were extracted and condensed by using protein extraction kits. And the intracellular and extracellular proteins were quantitatively detected on a micro-plate reader by using bicinchoninic acid (BCA) method. The proteins in cell culture fluid were qualitatively assayed by SELDI-TOF-MS. The results showed that the intracellular protein contents of LNCap cells were extremely higher than those of DU145 cells. After serum-free culture, both intracellular and extracellular protein contents of LNCap and DU145 were decreased to some extent. And the intracellular proteins were decreased by 5% in LNCap and by 36% in DU145 respectively, while the extracellular proteins were decreased by 89% in LNCap and 96% in DU145 respectively. SELDI assay revealed that there were 5 marker proteins in LNCap and 6 in DU145. Although both LNCap and DU145 cell lines originated from human prostate cancer, they had some differences in protein expression.
Cell Line, Tumor ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Mass Spectrometry/*methods ; Prostatic Neoplasms/*metabolism ; Proteins/chemistry ; Proteins/metabolism ; Proteomics/*methods ; Tumor Markers, Biological

Objective To study the antitumor effects on human prostate cancer cell lines of a traditional Chinese medicine named atractylodes macrocephala koidz volatile oil in vitro.Methods LNCaP and DU145 cell lines were normally cultured and were divided into 4 control groups including:Blank culture without serum and without cell (group A),Blank culture with serum but without cell (group B),LNCaP cell culture with serum (group C),DU145 cell culture with serum (group D).In the meanwhile,there were 6 experimental groups:adding 50 μg/ml of atractylodes macrocephala koidz volatile oil in culture of group C1 and group D1,250ug/ml in group C2 and group D2,and 500ug/ml in group C3 and group D3.All of the 10 groups were simultaneously cultured in 24-well-plates for 48 hrs,and each group was repeatedly studied for three times.Forty-eight hours later,every 2 × 106 cells of LNCaP or DU145 was seeded into each well and atractylodes macrocephala koidz volatile oil was added in 6 experimental groups,and saline water was added into 4 control groups.Another 48 hrs later,the culture solutions of the 10 groups were separately collected for testosterone (T),estradiol (E2),progesterone (P),vascular endothelial growth factor (VEGF),basic fibroblast growth factor (b-FGF),total prostate specific antigen (tPSA) and free prostate specific antigen (fPSA) analysis with enzyme-linked immunoassay kits.The experiment was repeated for 3 times,and the mean data were statistically analyzed by SPSS One-way ANOVA.Results The growth of cultured cells was found to have been effectively inhibited in group C1 and group D2.The inhibition severity of LNCaP cells was positively related with the drug concentration and time,while DU145 cells could only be highly inhibited (60.96％) after 24hrs drug treatments.In group C and group D,we found that both T were in very low level (0 ng/ml) whereas both E2 were in high levels (269 pg/ml and 239.81 pg/ml,P ＜ 0.05),no distinct differences showed in P; In addition,VEGF,b-FGF and fPSA were all in high values,whose values were 102.96 pg/ml and 1763.40 pg/ml,0.26 ng/ml and 6.41 ng/ml,0.16 ng/ml and 0.44 ng/ml,respectively; DU145 cells had higher values than LNCaP cells (P ＜ 0.05).As regard to the 6 experimental groups,in the groups C1,C2,C3 and D3,we found that T had been unexpectedly increased from 0 to 0.37 ng/ml (P ＜ 0.05),E2 continuously elevated from 239.81 pg/ml to 649.90 pg/ml (P ＜0.05),surprisingly P were also increased from 0.98 ng/ml to 9.83 ng/ml (P ＜0.01).On the contrary,VEGF,b-FGF and fPSA levels were all graduallydecreased,dropping down to 47.79 pg/ml and 59.56pg/ml,0 and 1.79 ng/ml,0 and 0.11 ng/ml,respectively; nevertheless,in group C2 and group D2,fPSA values were surprisingly increased from 0 and 0.04 ng/ml up to 1.78 ng/ml and 0.23 ng/ml respectively.Conclusions Atractylodes macrocephala koidz volatile oil has certain anti-tumor effects on human prostate cancer.Compared with LNCaP cells,DU145 cells have many different characteristics in sex hormone,cytokine and PSA expressions.

ObjectiveTo evaluate the application of multi-slice CT (MSCT) perfusion scan technique in predicting renal function recovery after unilateral hydronephrosis treatment.MethodsThirtyeight patients with unilateral obstructive hydronephrosis not shown on intravenous urography (IVU) and a normal contralateral kidney were recruited for this study.Patients were divided into detected (D) and undetected (UD) groups depending on whether the IVU detected urinary tract obstruction.All patients underwent plain abdominal X-ray,gray-scale ultrasonography,excretory urography and MSCT perfusion scan before and after the treatment.Patients were followed-up at six months or more after the treatment for a mean duration of 12.5 months (range from 6 to 22 ).ResultsOf the 38 cases,22 cases were in group D,16 cases were in group UD.On MSCT,renal cortex blood flow (BF) and blood volume ( BV ) value after treatment in group D were 561.1 ± 165.4 ml/( 100 g · min) and 35.9 ± 11.3 ml/100 g compared with before treatment rates of 361.6 ±109.7 ml/(100g· min) and24.1 ±10.2 ml/100g,t=-3.38,-2.34,P＜0.01,0.05.In the UD group,the differences of these parameters were after treatment 38.7 ± 15.4 ml/(100 g · min),10.306 ± 4.925 ml/100 g and before treatment 39.1 ± 22.5 ml/( 100 g · min) and 8.7 ± 4.4 ml/100 g,P ＞ 0.05.In the aspects of BF and BV,there were statistically significant differences between group D and group U D both before and after the treatment,t=9.09,4.15,P ＜ 0.01.ConclusionsM SCT perfusion can provide a valuable prediction technique of the renal function recovery in patients with unilateral obstructive hydronephrosis.Improvement of renal function can be expected after relief of obstructive hydronephrosis if the patients have a BF 361.6 ml/( 100 g · min) and BV 24.1 ml/100 g or greater measured by MSCT perfusion.

The differences in intracellular and extracellular protein expressions between human prostate cancer fines LNCap and DU145 were examined. The proteins of the two cell lines were extracted and condensed by using protein extraction kits. And the intracellular and extracellular proteins were quantitatively detected on a micro-plate reader by using bicinchoninie acid (BCA) method. The proteins in cell culture fluid were qualitatively assayed by SELDI-TOF-MS. The results showed that the intracellular protein contents of LNCap cells were extremely higher than those of DU145 cells. After serum-free culture, both intracellular and extracellular protein contents of LNCap and DU145 were decreased to some extent. And the intracellular proteins were decreased by 5% in LNCap and by 36% in DU145 respectively, while the extracellular proteins were decreased by 89% in LNCap and 96% in DU145 respectively. SELDI assay revealed that there were 5 marker proteins in LNCap and 6 in DU145. Although both LNCap and DU145 cell lines originated from human prostate cancer, they had some differences in protein expression.