Effects of Mage-H1 expression on PC12 cell differentiation and cell cycle

AIM: To explore the effects of melanoma-associated antigen H1 (Mage-H1) on cell proliferation and differentiation. METHODS: A phase contrast microscope was used to observe the morphological changes of PC12 cells treated with or without nerve growth factor (NGF). The expression of Mage-H1 in pre-and post-differentiated PC12 cells was detected by RT-PCR and Western blotting, and its potential effects on the cell cycle were analyzed by flow cytometry. RESULTS: After induced by NGF for 8 days, over 92% of PC12 cells were differentiated. The relative levels of Mage-H1 mRNA and protein in the differentiated PC12 cells were 4.6 times and 2.6 times higher than those in control cells,respectively. Moreover, the PC12 cells transiently expressed Mage-H1 were significantly arrested in G₀-G₁ phase as compared to the cells transfected with an empty vector.CONCLUSION: Mage-H1 inhibits the proliferation of PC12 cells and promotes the differentiation of the cells.     

Effects of tamoxifen on volume-activated Cl- currents of nasopharyngeal carcinoma cells at different stages of the cell cycle

AIM:To investigate the inhibitory effects of Cl- channel blocker, tamoxifen, on volume-activated Cl- currents of nasopharyngeal carcinoma cells (CNE-2Z cells) in G₁ and S phases. METHODS:Highly synchronous cells in G₁ phase and S phase were obtained by the serum starvation and the double-block techniques. The whole-cell patch clamp technique was used to observe the effects of tamoxifen on volume-activated Cl- currents and to analyze the anion permeability of volume-activated Cl- channels. RESULTS:47% hypotonic stimulation activated a Cl- current in the nasopharngeal carcinoma cells at the cell cycle stage G₁ phase and S phase. Tamoxifen at concentration of 10 to 30 μmol/L completely inhibited the current. However, the time needed to completely inhibit the current was dose-dependent and was different between G₁ phase and S phase. The time needed to completely inhibit the current was shorter in G₁ cells than that in S phase cells. The anion permeability sequence of the volume-activated Cl- channel was I->Cl->gluconate in both G₁ phase and S phase cells. The permeability of G₁ phase cells to I- was higher than that in S phase cells, but to gluconate was lower than that in S phase cells. CONCLUSIONS:The density of the volume-activated Cl- current, the anion permeability of the channel and the sensitivity of the current to tamoxifen were different between the CNE-2Z cells in G₁ phase and those in S phase. The results suggest that the expression of tamoxifen-sensitive, volume-activated chloride channels is differentiated at different stages of the cell cycle.     

Mcl-1 involves in G2/M arrest of HL-60 cells induced by diallyl disulfide

AIM: To investigate the role of Mcl-1 in the G₂/M arrest induced by diallyl disulfide (DADS) in leukemic HL-60 cells.METHODS: The inhibitory effect of DADS on human leukemic HL-60 cells was detected by CCK-8 method in vitro. Flow cytometry analysis was employed to observe the cycle arrest in HL-60 cells and the effect of DADS-induced G₂/M arrest on HL-60 cells with Mcl-1 gene knockdown by RNAi silencing. The expression of Mcl-1, PCNA and CDK1 in HL-60 cells treated with DADS was determined by Western blotting. The binding of Mcl-1 with PCNA and CDK1 was detected by coimmuno-precipitation. RESULTS: HL-60 cells were treated with DADS at concentration of 15, 30, 60, 120 or 240 μmol/L for 48 h. The inhibition rates of HL-60 cell proliferation were 31.15%, 55.88%, 66.14%, 75.29% and 80.35%, respectively, and gradually enhanced with the increase in the concentration of DADS (P<0.05). Flow cytometry analysis revealed that the proliferation of HL-60 cells was blocked by DADS in the G₂/M phase. Treatment with DADS for 24 h and 48 h at concentrations of 60 μmol/L and 120 μmol/L, the percentage of G₂/M phase cells increased as compared to the untreated cells (P<0.05). DADS induced arrest of HL-60 cells in G₂/M phase in a time- and dose- dependent manner (P<0.05). The results of Western blot analysis indicated that Mcl-1, PCNA and CDK1 in HL-60 cells were significantly reduced after treated with DADS (P<0.05). HL-60 cell cycle progression delayed by silencing Mcl-1 gene with siRNA technique, suggesting that silence of Mcl-1 gene led to G₂/M arrest. Compared to the cells treated with DADS only, the percentage of G₂/M cells raised in the cells with Mcl-1 gene silencing and treated with DADS (P<0.05), indicating that Mcl-1 gene silencing enhanced the effect of DADS-induced G₂/M arrest in HL-60 cells. The binding of Mcl-1 with PCNA and CDK1 was detected by coimmuno-precipitation and the formation of heterodimers was observed, which was decreased after treated with DADS for 4 h.CONCLUSION: DADS inhibits the proliferation of HL-60 cells and induces its G₂/M phase arrest. The decreased expression of PCNA is related to inhibiting the proliferation of leukemic cells. Knockdown of Mcl-1 gene enhances the effect of DADS-induced G₂/M arrest.     

Effect of norcantharidin on proliferation and invasion of human breast cancer cell line SKBR3 in vitro

AIM: To investigate the effect of norcantharidin（NCTD）on proliferation and invasion of human breast cancer cell line SKBR3 in vitro and its anticancer mechanisms.METHODS: MTT assay was used to determine SKBR3 cell proliferation. Light and FACScan were used to detect apoptosis and cell cycle. The invasiveness of SKBR3 was evaluated by the adhesion test，Matrigel experiment and the crossing-river test.RESULTS: NCTD had inhibitive effects on growth of SKBR3 cells in a dose and time-dependent manner, with the IC₅₀ value of 12.5 mg/L at 24 h．The cells treated with 10 mg/L NCTD for 24 h and 48 h showed typical apoptotic morphology and hypodiploid peak before G₁ phase. The cell cycle was arrested at G₂/M phase. The apoptosis percentage was up to 3.44% and 6.17%, and the G₂/M percentage was up to 35.82% and 38.70%. NCTD also could inhibit obviously the adhesion, movement and invasive capability simulating human basement membrane of SKBR3. Its effect was also in a dose-dependent manner. In the NCTD-treated group, crossing-river time was prolonged significantly and passing-membrane cells markedly decreased. CONCLUSION: NCTD in vitro inhibits not only the proliferation and growth of human breast cancer cells but also invasion and metastasis of the cells at relatively low concentration. NCTD shows prominent anti-tumor effects.     

Effects of crocetin on the apoptosis and the changes of its related regulating proteins caspase-3 and Bcl-2 induced by H2O2 in myocardial cells

AIM: To observe the effect of crocetin on the apoptosis and the changes of its related regulating proteins caspase-3 and Bcl-2 expression induced by hydrogen peroxide (H₂O₂) in cultured cardiomyocytes. METHODS: Changes of cellular morphology were detected under microscope. Apoptosis rates of the cells were analyzed by PI staining with flow cytometry. Expressions of caspase-3 and Bcl-2 proteins in the cells were determined by immunofluorescence with flow cytometry. RESULTS: In the concentrations used, more severe morphological changes with higher apoptosis rate of the cultured myocardial cells were seen in each H₂O₂ group than that in control group. When treated with 1×10-4 mol·L^－1 H₂O₂, the caspase-3 was increased and Bcl-2 protein decreased remarkably in the cells. But each dosage of crocetin, especially the highest one (5×10^－5 mol·L^－1, P<005 compared with 5×10^－7 mol·L^－1 group), seemed efficient in maintaining the cell morphology, reducing the cell apoptosis rate and improving the changes in caspase-3 and Bcl-2 protein expression in the cells exposed to 1×10^－4 mol·L^－1 H₂O₂. CONCLUSION: Crocetin obviously inhibits the apoptosis induced by H₂O₂ in the cultured myocardial cells. The mechanisms may involve the balance of the functions of the apoptosis-related regulating proteins, caspase-3 and Bcl-2 protein.     

Influence of human mutant p27 gene on the biological behaviors of colorectal cancer cells

AIM: To observe the influence of human mutant p27 gene (p27mt) on the growth and so as to investigate the function and mechanism of p27mt in gene therapy for colorectal cancer.METHODS: Colorectal cancer cell line SW480 was infected with recombinant replication defective adenovirus Ad-p27mt,and expression of p27mt protein was detected by Western blotting.The inhibitory effect of p27mt on SW480 and cell cycle were determined by flow cytometry,and DNA fragment was analyzed to identify the occurrence of apoptosis.RESULTS: After transfected with Ad-p27mt,p27 protein was highly expressed in SW480 cells.77.96% colorectal cancer cells were blocked in phase G₀/G₁,while in Ad-LacZ group and blank control group,27.57% and 25.29% cells were blocked in the same phase,respectively.Growth curve showed Ad-p27mt had an obviously inhibitory effect on the growth of SW480 cells.DNA fragment assay demonstrated that p27mt was able to induce the apoptosis of colorectal cancer cells.CONCLUSION: p27mt has an obvious blocking effect on colorectal cancer cell cycle,and most cells are blocked in phase G₀/G₁.This blockage is related with the growth inhibition and apoptosis induced by p27mt.     

Effects of bilirubin on oxygen free radicals and caspase-3 in acute lung injury rats

AIM:To investigate the mechanisms by which bilirubin inhibits acute lung injury (ALI). METHODS:30 female Wistar rats were divided into normal group, ALI group and bilirubin treatment group. ALI was induced by intravenous injection of LPS. The contents of OH^-, H₂O₂ and O₂^· in the lung as well as the expression of caspase-3 in the lungs were investigated. RESULTS:(1) The contents of OH^-, H₂O₂ and O₂^· in the lung homogenate and the expression of caspase-3 in the lungs in ALI group increased compared with those in normal group (P<0.05). (2) The contents of OH^-, H₂O₂ and O₂^· in the lung homogenate and the expression of caspase-3 in the lungs in bilirubin treatment group increased compared with those in normal group, but decreased compared with those in ALI models (P<0.05). CONCLUSION:(1) Bilirubin was shown to be able to ameliorate apoptosis in ALI rats. (2) The increase in the contents of OH^-, H₂O₂, O₂^· in ALI group indicated the development of oxidative lung injury, which was ameliorated by bilirubin. (3) Expression of caspase-3 confirmed that the model made by LPS was associated with apoptosis, which was reduced by bilirubin.     

Effect of vitamin K3 on apoptosis induced by androgen-independent prostate cancer cell PC-3M

AIM: To study the effect of vitamin K₃ (VK₃) on the induction of apoptosis in androgen-independent prostate cancer cell PC-3M in vitro.METHODS: Cell viability was estimated by MTT assay. AO/EB staining was performed to detect apoptotic cells. Apoptosis and the changes of cell cycle were detected by flow cytometry. NAC was used to observe the effect of growth inhibition by VK₃. RT-PCR was used to confirm the changes in gene expression. Levels of intracellular peroxides were estimated by using an oxidation-sensitive fluorescent probe DCFH-DA. RESULTS: PC-3M cells growth was significantly inhibited by VK₃ (≥60 μmol/L, P<0.05). The inhibitory effect was time and dosage dependent. The result of AO/EB staining showed that apoptosis of PC-3M cells were induced by VK₃. A typical subdiploid peak before G₀/G₁ phase was observed after treated for 12 h with VK₃ (60 μmol/L) by flow cytometry. The effect of growth inhibition treated with VK₃ was antagonized by antioxygen NAC (5, 10, 20, 40, 80 μmol/L). An increase in the level of DCF fluorescence after PC-3M cells were treated for 1-2 h with VK₃ was observed. Antioxidase GSH-Px and CAT were run-down after treated with VK₃. CONCLUSION: The results indicate that apoptosis in PC-3M cells is induced through oxidative stress by VK₃.     

Protective effects of pioglitazone on endothelial function in rats with hypercholesterolemia

AIM: To investigate the effects of pioglitazone,a PPARγ agonist,on endothelial cell (EC) dysfunction in hypercholesterolemic rats.METHODS: 36 healthy male Wistar rats were assigned to one of the following groups randomly (six rats in each group): control,hypercholesterolemia (HC),and HC treated with pioglitazone 1.5 mg·kg^-1·d^-1,3 mg·kg^-1·d^-1,10 mg·kg^-1·d^-1 and 20 mg·kg^-1·d^-1 (HC＋PIO),respectively.EC function was determined by comparing vasorelaxation to ACh,an EC dependent vasodilator,and acidified NaNO₂,an EC-independent vasodilator.Maximal positive and negative values of the instantaneous first derivative of LVP (+dp/dt_max and dp/dt_max) were determined by MS2000 system.RESULTS: (1) Hypercholesterolemia caused a significant endothelial diastolic dysfunction (maximal relaxation to ACh: 50.51%±2.45% vs 99.78%±3.01% in control,P<0.01).(2) Treatment with pioglitazone relieved EC-dependent vasodilatation in a dose dependent manner,and 10 mg·kg^-1·d^-1 is the best dose.(3) Pioglitazone not only improved EC function,but also reduced cardiac functional injury induced by hypercholesterolemia.CONCLUSION: EC dysfunction induced by hypercholesterolemia can be directly extenuated by pioglitazone,which may effectively prevent from subsequent atherosclerosis and ischemic heart disease.     

Double TdR blocking induces synchronization of cell cycle in human gastric cancer SGC-7901 cells

AIM: To investigate the effects of double thymidine deoxyribonucleoside (TdR) blocking on the cell cycle of human gastric cancer SGC-7901 cells.METHODS: SGC-7901 cells in the logarithm period were selected in the study and treated with TdR at concentration of 2 mmol/L, 4 mmol/L or 8 mmol/L for 15 h as the first blocking. After incubation in TdR-free medium for 10 h, the cells were treated with TdR at same concentrations again for another 15 h as the second blocking. The blocked cells were released by washing in fresh medium twice and incubation in TdR-free medium containing 10% fetal bovine serum for 12 h. The cells were collected and the cell cycle was detected by flow cytometry.RESULTS: By double TdR (2 mmol/L, 4 mmol/L and 8 mmol/L) blocking to synchronize the cell cycle, the cells in G₀/G₁ phase accounted for 77.3%, 77.5% and 77.0%, respectively. After further incubation for 12 h in TdR-free medium, the proportion of the cells in each phase of the cell cycles returned to normal range. CONCLUSION: The method of double TdR blocking is an ideal access in short term to acquire a large number of cells in G₀/G₁ phase.     

Apoptosis induction in gastric carcinoma cells by celecoxib combined with adriamycin

AIM:To study the apoptosis induction of cyclooxygenase-2 (COX-2) inhibitor，celecoxib and adriamycin (ADM) on tumor apoptosis of gastric carcinoma MGC-803 cells, and to explore their possible molecular mechanism(s) and interactions.METHODS:The number of MGC-803 cells was observed by MTT assay. Tumor apoptosis was studied by fluorescence microscopy, flow cytometry (FCM), and DNA ladder. RESULTS:MGC-803 cell number was significantly decreased with increasing dose of ADM. Cells were accumulated in G₀/G₁ phase and the number of cells in S phase was decreased. ADM (5 mg/L) combined with celecoxib (25 μmol/L) markably inhibited the growth of MGC-803 cells. Significant morphological changes of typical apoptosis were observed after treatment with combined use of celecoxib and ADM. Compared with ADM or celecoxib alone, ADM plus celecoxib obviously enhanced the DNA ladder fragment revealed by agarose gel electrophoresis of DNA. After exposure to combined celecoxib and ADM treatment for 48 h, MGC-803 cells were accumulated in G₀/G₁ phase. There was a decrease in the number of cells in S phase as compared to celecoxib or ADM alone. CONCLUSION:Celecoxib and ADM appear to have synergistic effects for the apoptosis induction. This may be an important prospect for applying COX-2 inhibitors to assist chemical therapy of ADM in clinical use.     

Effects of different doses of L-dopa on rotational behavior and amounts of cells expressing D2 receptors in hemiparkinsonian rats

AIM：To observe the effects of different doses of L-dopa on the rotational behavior and amounts of cells expressing D₂ receptors in striatum in hemiparkinsonian rats.METHODS：A hemiparkinsonian model was established in rats by pretreatment with 6-hydroxydopamine.The D₂ receptor expression were detected by immunohistochemical staining.The numbers of rotations induced by apomorphine was counted within 30 min before and after L-dopa (10 mg·kg^-1·d^-1,50 mg·kg^-1·d-^-1 or 100 mg·kg^-1·d^-1,ip) was introduced to Parkinson’s disease (PD) model rats for 15 days.RESULTS：In successful PD model rats,the increased percentage of D₂ receptor in lesioned side compared with intact side was associated linearly with the numbers of rotations within 30 min (r=0.927,P<0.01).After high dose of L-dopa intervention to PD model,the numbers of rotations decreased significantly (P<0.05),the amounts of cells expressing D₂ receptor at the lesioned side striatum decreased significantly (P<0.01).CONCLUSION：After high dose of L-dopa intervention,rotation behavior of PD rats improves,and D₂ receptor is down-regulated significantly.     

Effect of Yiqi Huoxue Jiedu Fang on the growth,metastasis and angiogenesis of Lewis lung carcinomas

AIM: To study the effect of Yiqi Huoxue Jiedu Fang (YHJ), composed of ginsenoside, penex notogingseng and berberin, on tumor growth and metastasis and to explore its mechanism.METHODS: Murine Lewis lung carcinoma transplant model was established and mice were treated with YHJ by intraperitoneal injection. After 10 days, the inhibitory rate of tumor, pathology of tumor and PCNA of tumor cells were detected. After 20 days, numbers of metastatic foci on lung surface and microvessel density (MVD) were determined. Expression of VEGF in tumor and serum were also analyzed by immunohistochemical test and ELISA, respectively. RESULTS: YHJ reduced the weight of tumor and the amount of metastatic foci. The inhibitory rates of tumor at high and low dose of YHJ (24 mg·kg_^-1·d_^-1, 12 mg·kg_^-1·d_^-1) were 48.29% and 37.26%, and the number of metastatic foci was 1.67 and 3.50, while control was 6.44. Furthermore, PCNA of tumor cells, MVD of tumor and VEGF expression in serum and tumor were decreased in YHJ treatment goup as compared with control. CONCLUSION: YHJ remarkably inhibits Lewis lung carcinoma growth and metastasis in mice. Its mechanism may be related to inhibition of angiogenesis.     

Astragali radix extract ameliorates renal resistance to atrial natriuretic peptide in rats with experimental nephrotic syndrome

AIM: To study the effects of astragali radix extract (ARE) on renal resistance to atrial natriuretic peptide (ANP) in rats with experimental nephrotic syndrome. METHODS: Male Sprague-Dawley rats were randomly divided into normal control, adriamycin nephropathy (ADR), ADR treated with ARE (2.5 g· kg^-1· d^-1) and ADR treated with benazepril (10 mg· kg^-1· d^-1). After 6 weeks, rats received intravenous infusion of 2% body weight isotonic saline. Urinary cGMP excretion (U_cGMPV), plasma ANP level, renal PDE5 activity and protein expression were also detected. RESULTS: ARE increased U_NaV while ACEI was not natriuretic. Nephrotic rats had a blunted natriuretic response and reduced rate of U_cGMPV after volume expansion despite higher plasma ANP concentration. ARE increased U_cGMPV and restored partly natriuretic response to volume expansion. The activity and protein abundance of renal PDE5 were high in nephrotic rats. ARE significantly reduced the PDE5 activity and protein expression. CONCLUSION: ARE may ameliorate the renal resistance to ANP in rats with adriamycin nephropathy by inhibiting the PDE5.     

Effect of salvianolic acid B on expression of TGF-β1 receptors and Smad2 in activated mesangial cells

AIM: To study the mechanisms of salvianolic acid B (Sal B)antagonizing mesangial cell activation and kidney fibrosis through investigating the effect of Sal B on expression of transforming growth factor-β₁ (TGF-β₁) receptors and Smad2 in TGF-β₁-stimulated renal mesangial cell activation. METHODS: Mesangial cells was isolated and purified from rat kidney. TGF-β₁ was used to establish rat primary mesangial cell activation model and Smad2,Smad7 protein expression was detected. Sal B (10^-6 mol/L and 10^-5 mol/L) was employed to treat the cells; α-smooth muscle actin(α-SMA) expression was analyzed by immunofluorescence staining and Western blotting. Mesangial cells were treated with Sal B alone or additional with TGF-β₁,and TGF-β₁ receptor Ⅰ (TβRⅠ),TGF-β₁ receptorⅡ (TβRⅡ),Smad2 phosphorylation and Smad2 protein expression was determined by Western blotting. RESULTS: Cell ular model was established by incubating with 5 μg/L TGF-β₁ for 24 h,and in early stage Smad2 was significantly phosphorylated. Sal B (10^-6 mol/L and 10^-5 mol/L) could inhibit α-SMA expression,which was the biomarker of activated mesangial cells. In addition,in Sal B group,the protein expression of TβRⅠand TβRⅡ was significantly down-regulated while Smad2 phosphorylation in mesangial cells was inhibited. CONCLUSION: Sal B inhibits the TGF-β₁-Smad pathway,the protein expression of TβRⅠ,TβRⅡ and Smad2 phosphorylation in mesangial cells,which is probably one of the mechanisms of Sal B alleviating kidney fibrosis.     

Lethal effect of VEGFR2shRNA on HL60 cells in vitro using lentiviral vector

AIM: To look for harmfulless anti-leukemia drug with selective high performance, lethal effect of small hairpin RNA (shRNA) on VEGFR₂ gene expression of tumor cell line HL60 in vitro.METHODS: The most effective VEGFR₂ siRNA was designed and screened. The shRNA oligo was designed and pU6/VEGFR₂ entry clone was constructed. HL60 was transfected transiently and vascular endothelial growth factor receptor 2(VEGFR₂) expression was tested with MTT assay, RT-PCR and Western blotting. The expression clone was constructed and cotransfected with ViraPowerTM Packaging Mix into 293FTTM cells to produce Lentiviral vectors harboring Lenti6/shVEGFR₂. The virion supernatant was added into HL60 cells and VEGFR₂ gene inhibitory effect was determined. RESULTS: The inhibitory rates of VEGFR₂ siRNA c were high. VEGFR₂ expression in HL60 was inhibited by using pU6/VEGFR₂ entry clone constructed with shRNA and pENTRTM/U6. For HL60 cells, the inhibitory rate was 84.9%. The expression of VEGFR₂ mRNA and protein decreased significantly. 48 hours after transfection of pU6/shVEGFR₂ entry clone and transduction of Lenti6/shVEGFR₂ expression clone, the cell inhibitory rates were similar. Cell growth inhibitory rate of entry clone descended rapidly after this time point, the expression clone changed slowly, reaching the peak at 96 hours, dropped slightly, having no significance deviation. CONCLUSION: in vitro, VEGFR₂ shRNA using lentiviral vector blocks VEGF/VEGFR₂ self-secretion in HL60 cells, which inhibits leukemia development.     

Effect of angelica on myocardial fibrosis post myocardial infarction in rats

AIM: To investigate 1) the role of transforming growth factor-β₁ (TGF-β₁) and macrophage infiltration during the development of myocardial fibrosis (MF) in rats after myocardial infarction (MI);and 2) mechanisms of MF post-MI and the inhibitory effect of angelica.METHODS: Sprague-Dawley (SD) rats were subjected to MI by ligating the left anterior descending coronary artery.The animals were randomly divided into three groups: sham, MI and MI+angelica.After 24 hours of ligation, rats received angelica (20 mL·kg^-1·d^-1, ip) or saline.Left ventricular hemodynamics were measured and rats were killed at week 1, week 2 and week 4, respectively.Collagen content, macrophage infiltration and TGF-β₁ expression were examined in the non-infarcted area.RESULTS: ① In MI group, the numbers of macrophage and TGF-β₁ expression were significantly upregulated compared to sham at week 1 post-MI and remained elevated at week 4 (P<0.01).Angelica significantly decreased macrophage infiltration and TGF-β₁ expression (P<0.01 vs MI).② Collagen content was increased significantly in MI group compared to sham at week 2 and week 4 (P<0.01), and decreased in MI+angelica group (P<0.05 vs MI).③ Cardiac function was markedly decreased post-MI in MI group (P<0.01), and improved at week 4 in MI+angelica group (P<0.05).CONCLUSION: In MF post-MI, angelica may have an antifibrotic effect by decreasing macrophage infiltration and TGF-β₁ expression, by which reactive myocardial fibrosis is reduced, and cardiac function is improved.     

Effect of G-CSF on dendritic cell subsets in donors mobilized peripheral blood cells

AIM: To explore the effect of granulocyte colony stimulating factor (G-CSF) on donor’s dendritic cell （DC）subsets in peripheral blood cells (PBC). METHODS: The subsets of dendritic cells in PBC were analyzed by CD34^-Lin^- HLA-DR⁺ cells and the levels of IL-12p40, IL-10, IFN-γ, IL-4 in the serum were tested by ELISA before (25 cases) or after G-CSF mobiling. The relationship between the ratio of DC₁/DC₂ (CD11c⁺CD123^-/CD11c^-CD123⁺) and CD34⁺/MNC was explored. RESULTS: CD34⁺/MNC in PBC harvests was above 0.4% in 23 cases, and the ratio of DC₁/DC₂ was lower, the HLA-DR expression of DC₂ was enhanced after G-CSF mobiling than before, but the levels of IL-12p40, IL-10, IFN-γ, IL-4 in donors serum and CD83 expression of DC₂ were not changed by G-CSF. CONCLUSION: DC₂ increased accompanied by the increase in CD34⁺ cells in the PBC harvests. Although the expression of HLA-DR in DC₂ was up-regulated by G-CSF, these DC₂ did not regulate Th2 cells to excrete inhibitor cell factors and kept on the precursor characters.     

DIM enhances effect of cis-diamminedichloroplatinum on growth inhibition and apoptosis in human prostate cancer cell PC-3

AIM: To study the effects of the combination of cis-diamminedichloroplatinum(DDP) and 3, 3-diindolylmethane (DIM) on the growth and apoptosis of human prostate cancer cell PC-3. METHODS: MTT method was applied to detect the cell growth inhibitory rate. The cell apoptosis was measured by the flow cytometry and acridine orange staining method. The expression of the anti-oncogene p21 was detected by RT-PCR technique. RESULTS: The combination of 60 μmol·L^-1 DIM and 0.4 mg·L^-1 DDP effectively inhibited the growth and induced apoptosis in PC-3 cells. This result was the same as the effect of using 4 mg·L^-1 DDP only. The cell growth inhibitory and apoptosis rates for the combination of DIM and DDP were much higher than those for the individual effect. Both the combination and the single effect of these two medicines (i.e., DIM and DDP) all strengthen p21 mRNA expression significantly, and the effect of combination was more significant. CONCLUSION: DIM significantly enhances the effects of DDP on the growth inhibition and apoptosis induction in PC-3 cells.