Effect of repeated measles vaccine injection on interleukin-12,interleukin-13 levels in asthmatic children

AIM and METHODS:To study the immunological effect of measles vaccine therapy on asthmatic children, we examined the changes of interleukin-12 , interleukin-13 and total serum IgE levels in plasma and cultured peripheral blood mononuclear cells(PBMC) supernatant by means of ELISA in 13 mild-moderate asthmatic children treated with measles vaccine. Results were compared with 12 anti-symptomatic treatment mild-moderate asthmatic children and 17 normal children control group.RESULTS:After measles vaccine treatment, IL-13 and total serum IgE levels decreased remarkably, statistically lower than that of group receiving only anti-symptomatic treatment. There was no statistical difference in IL-12 level between the two group. Correlation analysis: 1)IL-12 level of plasma was negatively correlated to the level of serum total IgE, there was no correlation of supernatant IL-12 in PBMC to the total serum IgE; 2)IL-13 levels in plasma and PBMC were positively correlated to the level of total serum IgE; 3) IL-12 level was negatively correlated to IL-13.CONCLUSION:Measles vaccine could down-regulate IL-13 level,therefore decrease total IgE synthesis,but not affect IL-12 level in asthmatic children.     

IgE-FcεRI crosslink accelerates atherosclerosis development in allergic asthmatic mice

AIM:To investigate whether allergic asthma accelerates the development of atherosclerosis in mice related to Th2 cells and interleukin-4 (IL-4), and the roles of activation of macrophages by immunoglobulin E (IgE)-Fc ε receptor I (FcεRI) crosslink during the process. METHODS:Six-week-old ApoE^-/- mice were sensitized and challenged by ovalbumin to establish the allergic asthma model, and then assigned to 3 groups:control group, asthmatic placebo group and asthmatic IL-4 monoclonal antibody (mAb) intervention group (intervention for 8 weeks). The lesion area was measured by oil red O staining. The percentages of Th2 cells in the splenocytes of the mice were analyzed by flow cytometry. The mRNA expression of IL-4 and the macrophage-related inflammatory factors, monocyte chemotactic protein 1 (MCP-1), macrophage inflammatory protein-1α (MIP-1α) and IL-6, in the spleen was detected by real-time PCR. Local IgE and FcεRIα expression in the plaque was evaluated by immunofluorescence/immunohistochemical staining, and the circulating IL-4 and IgE were measured by ELISA. RESULTS:Accompanied by aggravated atherogenesis in asthmatic ApoE^-/- mice, the proportion of Th2 cells and IL-4 mRNA in the spleen, IgE and FcεRIα expression in the aortic root, and the mRNA expression of MCP-1, MIP-1α and IL-6 were markedly increased. After 8-week treatment with IL-4 mAb, the lesion area in the aortic root of asthmatic ApoE^-/- mice was markedly decreased, the elevated IgE and FcεRIα expression was significantly decreased, and the mRNA expression of macrophage-related inflammatory factors was also decreased. CONCLUSION:Allergic asthma accelerates the atherosclerosis in ApoE^-/- mice, which is associated with the increased Th2 cells and IL-4, and the activation of macrophages by IgE-FcεRI crosslink.     

Expression of PI3K in airway smooth muscle cell of asthmatic rats

AIM: To investigate the expression of PI3K in airway smooth muscle cell (ASMC) of asthmatic rats.METHODS: 16 Wistar rats were divided into two groups, asthma and normal control at random. After establishment of asthmatic model, flow cytometry, immunofluorescence and Western blotting were applied to detect the growth fraction of ASMC and the expression of PI3K in cultured ASMC from each rat.RESULTS: It was revealed from flow cytometry that the ratio of S + G2/M to total number of cells in asthma group ［ (27.90±3.44) % ］ was higher than that in normal control group ［ (13.00±1.56) %, P<0.05］. The expression of PI3K was observed in both asthma and normal control group. However, it was much higher in asthma group than that in normal control group. There was a positive correlation between the expression of PI3K and the growth fraction in ASMC. CONCLUSION: The increased expression of PI3K might play an important role in regulating the proliferation of ASMC in asthma.     

Construction and expression analysis of idiotype TCR Vβ2 DNA vaccine in vitro

AIM:To develop an anti-lymphoblastic leukemia TCR idiotypic DNA vaccine, analyze its transfer activity into K562 cells and to detect its expression in vitro. METHODS:The TCR Vβ2 gene segment, which was identified from an idiotypic TCR Vβ2 clone-Molt4 cell line, was amplified using RT-PCR, and the PCR products were then cloned into pIRES vector. The recombinant plasmids were transferred into K562 cells. The condition of idiotypic protein expression was tested by indirect immunophenotyping fluorescein dyeing, SDS-PAGE and Western blotting. RESULTS:The recombinant DNA plasmid, pIRES-TCR Vβ2, was developed successfully. The expression of TCR Vβ2 was identified on the surface of K562 cells. A 15 kD protein, which bound to TCR Vβ2 antibody specifically, were identified from pIRES-TCR Vβ2 transfected K562 cells by Western blotting, indicating that TCR Vβ2 protein was expressed in vitro. CONCLUSION:The recombinant plasmid pIRES-TCR Vβ2 DNA vaccine was developed successfully, which was expressed TCR Vβ2 protein specifically in transfected K562 cells.     

Effect of estrogen receptor 2 on the cell proliferation in prostate cancer cell line PC-3M

AIM: To construct the recombination plasmid pcDNA3.1-hERβ with the human estrogen receptor 2 (ESR2) full length cDNA and transfect it into hormone-independent prostate cancer PC-3M cell line, and to study the effects of ESR2 on proliferation in transfected cells. METHODS: The complete cDNA of ESR2 was obtained from human ovary tissue by RT-PCR technique and cloned into eukaryotic expression vector pcDNA3.1 by using gene recombination technique to construct the pcDNA3.1-hERβ recombination plasmid. The plasmid was detected by endonuclease digestion and DNA sequencing and was transfected into PC-3M cells. MTT and FCAS assay were used to test the effects of ESR2 on the ability of proliferation in PC-3M cells. RT-PCR and Western blotting were used to detect the expressions of cyclinD1 and P21Cip1. RESULTS: The results of sequencing and endonuclease digestion demonstrated that the construction of pcDNA3.1-hERβ recombination plasmid was successful. The sequence analysis suggested that the ESR2 sequence detected by PCR was identical to that published in GenBank, and the product of endonuclease was as long as the complete human ESR2 gene. 48 h after transfected the pcDNA3.1-hERβ into PC-3M cells, the expression of ESR2 mRNA and protein levels increased significantly detected by RT-PCR and Western blotting. Compared to the cells transfected with vector as control, the PC-3M cells transfected with pcDNA3.1-hERβ showed that cell population decreased and proliferation activity degraded. FCAS showed that the cells in G0/G1 stage increased and in S stage or G2/M stage decreased. RT-PCR and Western blotting showed that the expression of cyclinD1 gene reduced and expression of P21Cip1 increased. CONCLUSION: The recombination of plasmid pcDNA3.1-hERβ is constructed and transfected into the PC-3M cells successfully. The activity of cell proliferation is inhibited after pcDNA3 transfection.1-hERβ. It is possible that ESR2 inhibits cell proliferation by the expression of proliferation related genes cyclinD1 and P21Cip1.     

Construction of fusion expression vector AR-GFP and its expression in Hek293 cells

AIM: To construct a hAR and GFP fusion gene vector and to observe the AR-GFP gene expression in Hek293 cells. METHODS: A recombined vector pcDNA3.1/myc-HisA-AR-GFP (pH-AG) was constructed by gene engineering technique. The recombined vector was transfected into Hek293 cells using calcium phosphate. RESULTS: AR-GFP fusion protein was successfully expressed in Hek293 cells without biologic activity, which was confirmed by fluorescence microscopy and Western blotting. CONCLUSION: The Hek293 cells transfected by AR-GFP fusion gene can express its protein successfully. However, it is not a cellular model for ARI screening.      

Up-regulation of Dicer1 gene expression by prostate-specific transcriptional factor NKX3.1

AIM: To investigate the effect of NKX3.1 on the Dicer1 gene expression.METHODS: The NKX3.1 eukaryotic expression plasmid was transfected into PC3 cells. The stable clones were isolated using cloning cylinders and grew continuously under G418 selection. The gene expression profile in PC3 (+) cells induced by NKX3.1 was analyzed by cDNA microarray. The effect of NKX3.1 on the Dicer1 expression was further investigated by RT-PCR and Western blotting in PC3 and PC3 (+) cells according to the results of gene chip. To determine if the increase in Dicer1 promotes the mature of microRNA, the pMIR-report luciferase expression plasmid of miRNA let-7a1 target sequence (pMIR-report-let7a1T) was constructed and transfected into PC3 and PC3 (+) cells. The effect of the miRNA let-7a-1 on its target sequence was determined by luciferase reporter assay.RESULTS: The result of gene chip showed that the expression level of Dicer1 gene was higher in PC3 (+) cells than that in PC3 cells. The results of RT-PCR and Western blotting indicated that the expression of Dicer1 gene was much higher in PC3 (+) cells than that in PC3 cells. The relative luciferase activity was much lower in PC3 (+) cells than that in PC3 cells when the cells were transfected with the pMIR-report-let7a1T vector.CONCLUSION: Up-regulation of Dicer1 expression induced by NKX3.1 promotes the mature and functions of microRNAs in prostate cancer PC3 cells.     

Effects of dexamethasone on intracellular expression of TH1/TH2 cytokines in experimental asthma mice

AIM: To study the effects of dexamethasone (DXM) on intracellular expression of T_H1/T_H2 cytokines and the mechanism of that during the development of asthma. METHODS: Eighteen BALB/c mice were divided into 3 groups at random: control group, asthma group, and DXM treated group, with 6 mice in each. The expressions of T-box expressed in T cells (T-bet) and GATA-binding protein-3 (GATA-3) in lung tissue were detected by Western blotting. The expressions of intracellular cytokines interleukin-4 and interferon-γ in CD4⁺ T cell were measured by flowcytometry.RESULTS: The results of flow cytometry indicated that the ratio of intracellular cytokines IL-4/IFN-γ in CD4⁺ T cells in asthma group was much higher than that in control group (P<0.01), the ratio of intracellular IL-4/IFN-γ in T cells in DXM group was lower than that in asthma group significantly (P<0.01). The expression of T-bet in lung tissue in asthma group was lower than that in control group significantly (P<0.01), while GATA-3 was higher than that in control group significantly (P<0.01). The expressions of T-bet and GATA-3 in DXM group were much lower than those in asthma group (P<0.01), but the decreased degree of GATA-3 was more than that of T-bet. CONCLUSION: With pathological process of asthma, to reverse the ratio of IL-4/IFN-γ in CD4⁺ T cell by regulating T-bet and GATA-3 expression can improve the inflammatory reaction and may be one of the mechanisms of DXM in treating asthma.     

Role of GATA-3 in the pathogenesis of airway inflammation in a rat asthma model

AIM: To investigate the role of GATA-3 in the pathogenesis of airway inflammation in a Wistar rat asthma model. METHODS: The Wistar rat asthma model was made with conventional method and animals were divided into five groups (10 rats in each group): asthma group (A group), dexamethasone group (D group), antisense oligonucleotide group (AS group), nonsense oligonucleotide group (NS group) and normal control group (N group). Antisense, nonsense oligonucleotide were administered intranasally, and the dexamethasone was injected intraperitoneally. The airway inflammation was observed with HE staining method. The GATA-3 positive cells were stained immunohistochemically. The GATA-3 mRNA expression in pulmonary tissue was investigated with RT-PCR. The GATA-3 protein in pulmonary tissue was detected by Western blotting. RESULTS: In contrast to N group, the expression of GATA-3 mRNA, protein and the amount of inflammatory cells in pulmonary tissue in group A were increased significantly (P<0.01) and were decreased evidently in group AS and D (P<0.01). The expression of GATA-3 mRNA, protein and the amount of inflammatory cells in NS group were obviously increased compared with those in gropu AS and D (P<0.01). The expression of GATA-3 was related to the amount of eosinophils (r=0.995). CONCLUSION: GATA-3 antisense oligonucleotide blocks the expression of GATA-3 gene and the infiltration of eosinophils. GATA-3 plays an important role in the effector phase of allergic airway inflammation in a Wistar rat asthma model.     

Metabolic mechanism of retinoic acid in glioma cells and its impact on cell proliferation

AIM: To investigate the mechanism for regulating the synthesis and metabolism of retinoic acid in glioma cell line SWO-Z2 and its effect on cell proliferation. METHODS: The siRNA targeting to human KLF9 mRNA was transfected into SWO-Z2 cells. The silencing efficiency was detected by real-time PCR and Western blotting. After silencing of KLF9 , the protein level of ALDH1A1 was detected by Western blotting. CCK-8 colorimetric assay was used to screen the optimal concentration of retinoic acid, and the strongest inhibitory effect of retinoic acid from 3 types of chemicals,13- cis -retinoic acid (13- cis RA),9- cis -retinoic acid (9- cis RA)and all- trans retinoic acid (ATRA), on SWO-Z2 cell growth was selected. Western blotting was also applied to explore the expression levels of cyclin D1, Bcl-2, cleaved PARP and GFAP in SWO-Z2 cells with the treatment of ATRA for 72 h. Simultaneously, the mRNA levels of retinoic acid receptors (RARs) in SWO-Z2 cells were determined by real-time PCR. RESULTS: siRNA-KLF9 knocked-down the expression of KLF9 and down-regulated the expression of aldehyde dehydrogenase 1 family member A1 (ALDH1A1) at mRNA and protein levels (P<0.05). Among the 3 retinoic acid drugs, ATRA was the most effective in inhibiting the proliferation of SWO-Z2 cells. After treated with ATRA on SWO-Z2 cells for 72 h, the expression of cleaved PARP was increased, Bcl-2 and cyclin D1 were decreased, and GFAP didn't change. The mRNA level of RARs in SWO-Z2 cells was very low. CONCLUSION: KLF9 positively regulates the expression of ALDH1A1 gene to increase the synthesis of retinoic acid. ATRA inhibits proliferation but does not induce differentiation of SWO-Z2 cells, which might result from lack of retinoic acid receptors in human glioma cells.     

Effects of adiponectin cDNA transfection on glycogen synthesis and glucose oxidation in myotubes

AIM: To study the effects of adiponectin on glycogen synthesis and glucose oxidation in C2C12 myotubes. METHODS: Plasmid pcDNA3.0-mad with cDNA of mouse adiponectin, and vector pcDNA3.0 were transfected into C2C12 cells by lipofectAMINE 2000 reagent, respectively. Stably transfected cells were screened by 500 mg/L G418 for 3 weeks. Adiponectin protein expression was determined by Western blotting analysis and immuno-histochemistry. Glucose oxidation and glycogen synthesis detections were divided into control, vector and pcDNA3.0-mad (mad) group. Each group was further divided into 4 subgroups with 0, 0.5 nmol/L, 5 nmol/L or 100 nmol/L insulin (n=6), respectively. Detection of glucose oxidation and glycogen synthesis was carried out with ［14C］-labeled glucose by counting radioactivity of ［14CO2］ or ［14C］ labeled glycogen with scintillation, respectively. RESULTS: Adiponectin protein expression was only detected in the mad group by either Western blotting analysis or immunostaining. The rate of glucose oxidation increased more with the elevation of insulin concentration in the mad group than that in other 2 groups: the regression coefficient of control, vector and mad group was 23.34， 23.23 versus 26.06, respectively. No significant difference in either basic or insulin-stimulated glucose oxidation and glycogen synthesis between mad group and the other two groups was observed (P>0.05). CONCLUSION: Transfection with adiponectin cDNA has no significant effect on the glucose oxidation and glycogen synthesis in C2C12 myotubes.     

Inhibitory effects of FRNK on collagen synthesis in hepatic stellate cells

AIM:To investigate the effect of focal adhesion kinase-related non-kinase (FRNK) on collagen synthesis in hepatic stellate cells (HSCs).METHODS:HSCs were transfected with FRNK by cationic liposome method. The expressions of FRNK at the protein level in HSCs were detected by Western blotting analysis. HSCs collagen synthesis capability was examined by［3H］-Pro incorporation assay.RESULTS:The exposure of HSCs to FRNK led to the up-regulated expression of FRNK protein, and it was at 48 h after transfection that the FRNK protein content was the highest. Moreover, after FRNK was transfected successfully in HSCs, the total collagen synthesis and type I collagen synthesis were inhibited.CONCLUSION:After FRNK is transfected successfully in HSCs using Lipofectamine, the synthesis of total and type I collagen in HSCs is inhibited.     

Effect of mfn2 on mitochondrial function in steatosis hepatocytes

AIM: To investigate the effect of mfn2 on mitochondrial function in steatosis hepatocytes. METHODS: Plasmid pEGFP-mfn2 was transfected into hepatocyte strain L02 by Lipofectamine 2000 in vitro, then the steatosis model of hepatocytes was establish by oleic acid induction. RT-PCR was used to evaluate mRNA expression and Western blotting was use to detect the protein expression. ATP level was determined by firefly luciferase bioluminescent. ROS production was measured by fluorescence probe DCFH-DA. Chondrosome transmembrane potential of L02 was observed by labeling of JC-1 and FCM. RESULTS: The stable expression of ectogenesis mitofusin2 in L02 cells was confirmed by RT-PCR and Western blotting. In the model of oleic acids induced lipid formation, Mfn2 obviously inhibited the descent of chondrosome transmembrane potential and ATP level, and increased ROS production in L02 cells. CONCLUSION: Up-regulated expression of mfn2 attenuates mitochondria dysfunction caused by oleic acids induced lipid formation.     

Role of translationally controlled tumor protein in PRL-3-promoted metastasis of colon cancer cells

AIM: To study the role of translationally controlled tumor protein (TCTP) in the proliferation, migration and invasion of phosphatase of regenerating liver-3(PRL-3)-promoted colon cancer cells.METHODS: The vectors pAcGFP-C3 and pAcGFP-C3-PRL-3 were constructed and transfected into the colon cancer cell line LoVo.LoVo-PRL-3 cells stably expressing PRL-3 and LoVo-control cells were established. The expression levels of PRL-3 and TCTP in both cells were detected by Western blotting and real-time PCR. The specific siRNA sequence for TCTP mRNA and control-siRNA were synthesized and transfected into the LoVo-PRL-3 cells. TCTP expression at mRNA and protein levels in LoVo-PRL-3 was detected by Western blotting and real-time PCR 24 h, 48 h and 72 h after transfection. The proliferation, migration and invasion abilities of LoVo-control cells, LoVo-PRL-3 cells, TCTP-siRNA and control-siRNA cells were detected by CCK-8 assay and the method of Transwell cell culture chambers.RESULTS: The expression of TCTP at mRNA and protein levels in LoVo cells was significantly increased after PRL-3 transfection (P<0.05). TCTP mRNA was significantly inhibited 24 h, 48 h and 72 h after transfection of TCTP-siRNA (P<0.01). TCTP protein was also significantly inhibited 48 h and 72 h after transfection (P<0.01). Compared with LoVo-control cells, the proliferation, migration and invasion abilities of LoVo-PRL-3 cells were significantly enhanced (P<0.05). However, lowering the up-regulated expression of TCTP in LoVo-PRL-3 cells inhibited the proliferation, migration and invasion abilities (P<0.05). CONCLUSION: PRL-3 promotes proliferation, migration and invasion of colon cancer cells by up-regulating the TCTP expression. siRNA targeting TCTP may be an effective method for prevention and treatment of colon cancer cell metastasis.     

p38MAPK expression and apoptosis in the rat unilateral ureteral obstruction model

AIM: To investigate the expression of p38 mitogen-activated protein kinase (p38MAPK) in the kidney after unilateral ureteral obstruction (UUO) in rats and the functional role of it on apoptosis and fibrosis.METHODS: Eighteen Wistar rats underwent UUO were killed at 3, 7, 14 days. Additional 7 rats were sham operated. Histological changes were observed by HE and Masson staining. Immunohistochemistry study was performed on renal tissue for proliferating cell nuclear antigen (PCNA). Apoptotic cells were determined by terminal deoxynucleotidyl transferase- mediated dUTP nick end labeling (TUNEL) and the electrophoresis analysis of genomic DNA. Western blotting of cysteinyl aspartate specific proteinase-3 (caspase-3), p38MAPK and p-p38MAPK were measured.RESULTS: UUO induced a significant increase in renal tubular and interstitial cell apoptosis, immunohistochemistry of PCNA and Western blotting of caspase-3, p-p38MAPK as well as severe morphology changes. However, there was no significant difference between UUO and the control in Western blotting of p38MAPK.CONCLUSION: An in vivo model of renal fibrosis after UUO demonstrates that activated or phosphorylated p38MAPK plays a role in apoptosis of renal tubulointerstitial cells.     

Effects of Foxp3 transfection on splenocyte functions in asthma mice

AIM: To study the expression and the effects of Foxp3 on the immunologic functions by transfecting the Foxp3 eukaryotic expression plasmid into the splenocytes of the asthma mice. METHODS: The mice were sensitized and challenged by ovalbumin to make asthma model. The splenocytes were harvested and cultured. The Foxp3 expression vector pcDNA3.1(-)-Foxp3 was transfected into the splenocytes with electroporation. The splenocytes transfected with empty vector and control splenocytes (non-transfected) were also set up. The expression of Foxp3 at mRNA and protein levels was detected by RT-PCR and Western blotting, respectively. The proportion of CD4⁺CD25⁺ Treg cells/CD4⁺ cells was measured by flow cytometry. Proliferation of the splenocytes was analyzed with MTT assay. ELISA was used to determine the levels of interleukin 4 (IL-4) and interferon γ (IFN-γ) in the supernatant of the splenocytes. RESULTS: The expression of Foxp3 at mRNA and protein levels in transfection group was significantly higher than that in empty vector group and control group. The proportion of CD4⁺CD25⁺Treg cells/CD4⁺ cells in transfection group was higher than that in empty vector group and control group. The proliferation of transfected cells was markedly inhibited compared with empty vector group and control group. The levels of IL-4 and IFN-γ were significantly lower in transfection group than those in empty vector group and control group. CONCLUSION: The transfected Foxp3 gene overexpresses in the splenocytes of asthma mice. Foxp3 increases the number of CD4⁺CD25⁺ T cells and inhibits the proliferation and production of Th1/Th2 cytokines in splenocytes.     

13-MTD induces apoptosis of MCF-7 breast cancer cells by activating MAPK pathway and inhibiting Akt signaling

［ABSTRACT］AIM: To study the effect of 13-methyltetradecanoic acid (13-MTD), a saturated branched-chain fatty acid, on apoptotic induction in breast carcinoma cell line MCF-7 and its underlying mechanisms. METHODS: Breast carcinoma cell line MCF-7 and normal breast epithelial cells MaEC were treated with solvent or 13-MTD at concentration of 140 mg/L. Apoptosis was analyzed by flow cytometry. Phosphorylation of JNK, p38, FADD and Akt after treated with 13-MTD were detected by Western blotting. RESULTS: 13-MTD effectively induced apoptosis of breast carcinoma cell line MCF-7 and no influence to normal breast epithelial cells MaEC, which were confirmed by flow cytometry analysis, was observed. The results of Western blotting showed that obvious increase in p38 and JNK phosphorylation. No significant difference of FADD phosphorylation was observed. However, evidently decrease in Akt phosphorylation was found after treated with 13-MTD. CONCLUSION: 13-MTD was a new safe, effective chemotherapeutic drug. Its underlying mechanisms are through activating MAPK pathway and inhibiting Akt pathway to induce the cancer cells apoptosis.     

Effect of rhEGF activating extracellular signal-regulated kinase on the expression of Bcl-2 and p53 protein in human amniotic cells

AIM: To explore the effect of recombinant human epidermal growth factor (rhEGF) on the ERK1/2 pathway and its safety in use when the proliferation of cultured FL cells (human amnion epithelial cells) were promoted by rhEGF.METHODS: FL cells were stimulated by rhEGF at various concentrations.The proliferation rate of FL cells was evaluated by MTT assay.Western blotting was used to detect phosphorylation of ERK1/2 (p-ERK1/2) and the change of Bcl-2 and P53 protein.RESULTS: The proliferation rate FL cells reached the maximum when the concentration of rhEGF was 10 μg/L (42.4％，P<0.01).Western blotting showed that the activity of p-ERK1/2 increased significantly when rhEGF ranged in 10 μg/L to 60 μg/L.Level of Bcl-2 was unchanged in each groups.Level of P53 decreased significantly when the concentration of rhEGF was 60 μg/L.CONCLUSION: It is safe when rhEGF presents in optimization concentration of 10 μg/L.The capability of apoptosis may be depressed when rhEGF is in larger dose.     

Establishment of human pancreatic tumor xenograft mouse model for evaluating tumor-homing and gene-silencing effects of siRNA-loading nanoparticles

AIM：To establish an effective and rapid method to develop transplanted subcutaneous pancreatic carcinoma by inducing PANC-1 cells into nude mice, and then use this mouse model to evaluate the tumor-homing and gene-silencing effects of siRNA-loading nanoparticles in vivo. METHODS：Different numbers of PANC-1 cells in 100 μL or 300 μL PBS were inoculated subcutaneously into the right flank of BALB/c (nu/nu) mice. When the tumor volume reached 100 mm3, siRNACY5.5 nanoparticles were injected through the mouse tail vein to perform in vivo imaging assay. Besides, the mice were randomly divided into 3 treatment groups treated with PBS, scrambled control RNA nanoparticles and siKras nanoparticles, respectively. The protein expression of Kras was detected by Western blotting and immunohistochemical staining. RESULTS：After inoculated with 1×107 PANC-1 cells in 300 μL PBS, all mice developed tumors within 2 weeks. The in vivo results showed that siRNA-loading nanoparticles accumulated in the tumor tissues and exerted gene silencing effect. CONCLUSION:In the present study, an effective and rapid method was established for PANC-1 cells to induce transplanted subcutaneous pancreatic carcinoma in nude mice within 2 weeks, which is suitable for in vivo imaging and treatment evaluations as a reproducible and reliable way for the further experiments.