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1.
Chinese Pharmacological Bulletin ; (12): 1012-1019, 2018.
Article in Chinese | WPRIM | ID: wpr-705168

ABSTRACT

Aim To investigate the anti-proliferation effect of honokial on human colon cancer cells HCT116 and the possible mechanism. Methods Crystal stai-ning and flow cytometry assay were introduced to test the proliferation inhibitory and cell cycle arrested effect of honokial on HCT116 cells. Western blot was used to analyse the expression of PCNA and induction effect of apoptosis. Western blot and PCR assay were conducted to assess the change of BMP7. Via exogenous adenovi-ruses of BMP7 and its antibody combined with honoki-al,crystal violet staining and Western blot were used to analyse the effect on HCT116 cells. Results Honoki-al inhibited the growth of HCT116 in a time-and con- centration-dependent way,induced apoptosis and arres-ted at G2 phase; Western blot assay showed honokial up-regulated the level of PCNA and BMP7,and down-regulated the expression of Bcl-2; Western blot result also showed that exogenous adenoviruses of BMP7 en-hanced the effect of honokial on proliferation inhibition and apoptosis, while BMP7 antibody reversed such effects of honokial. Conclusion Honokial can inhibit the proliferation and induce apoptosis on HCT116 cells,which may be mediated by the up-regulation of BMP7.

2.
Acta Pharmaceutica Sinica ; (12): 44-49, 2014.
Article in Chinese | WPRIM | ID: wpr-297973

ABSTRACT

This study is to investigate the anti-tumor activities of a novel cyclophosphamide derivate 4, 6-diphenyl cyclophosphamide (9b) in vivo and in vitro, and its possible mechanism of action. The inhibitory effects of 9b on human hepatoma cell line HepG2, human breast carcinoma cell line MCF-7 and human myeloid leukemia cell line K562 were measured by MTT assay in vitro. Cell cycle distribution and apoptotic rate were evaluated by flow cytometry. To evaluate the anti-tumor effect of 9b in vivo, mouse model bearing inoculated H22 tumor was established. The results indicated that 9b could inhibit the proliferation of HepG2, MCF-7 and K562 cells in a dose and time dependent manner. The ICo50 values of 9b were 32.34 micromol.L-1 to HepG2 cells, 87.07 micromol.L-1 to MCF-7 cells and 149.10 micromol.L-1 to K562 cells after incubation for 48 h. The results of flow cytometry indicated that after being treated for 48 h with different concentrations of 9b, the ratios of HepG2, MCF-7 cells at the Go/G1 phase and K562 cells at the G0/Gl phase and G2/M phase increased significantly compared with control group, and the apoptotic rate increased with the increase of the concentration of 9b. 9b could significantly reduce tumor weight of H22 solid tumor mouse model in vivo. To summarize, 9b showed significantly anti-tumor activity in vivo and in vitro, of which the mechanism might be associated with the change of cell cycle distribution and induction of tumor cell apoptosis.


Subject(s)
Animals , Antineoplastic Agents, Alkylating , Chemistry , Pharmacology , Apoptosis , Cell Cycle , Cell Line, Tumor , Cell Proliferation , Cyclophosphamide , Chemistry , Pharmacology , Dose-Response Relationship, Drug , Female , Humans , Inhibitory Concentration 50 , Liver Neoplasms, Experimental , Pathology , Male , Mice , Molecular Structure , Random Allocation , Tumor Burden
3.
Article in Chinese | WPRIM | ID: wpr-642291

ABSTRACT

Objective To observe the metabolism and distribution of arsenic in liver and brain of offspring rata by exposure to arsenic of pregnant rats or lactation dams and weaned pups,and explore if arsenic could penetrate the placental barrier,lactation barrier and blood brain barrier. Methods The Wistar female rots were randomly divided into four groups according to body weights,12 in each group,and were fed with drinking water that contained arsenic(NaAsO_2) 0,10,50,100 mg/L beginning from the gestafional day 6 until pups 42 days old. Pups were separately sacrificed on postnatal day(PND) 0,15,28,42. Arsenic in liver and brain of offspring rots and in breast milk was examined by atomic absorption speetrophotometer with an arsenic speeiation pretreatment system. Results Concentration of iAs,MMA,DMA of brain in 50,100 mg/L groups were higher than that of 0 mg/L group[0,0,0,(7.3±6.6),0,(44.2±27.4)ng/g]on PND 0,42[iAs: (120.0±46.0),(195.5±125.3),(216.5±278.4),(176.6±151.8) ng/g; M MA: (47.2±18.1),(199.6±389.1),(47.4±55.2),(82.7±79.2) ng/g; DMA: (984.3±377.4),(2222.1±1433.2),(998.1±368.3),(1781.3±715.7)ng/g,all P < 0.05]. Concentration of DMA of brain in 50,100 mg/L groups were higher than that of 0 mg/L group[(13.9±18.1),(50.6±98.3)ng/g]on PND 15,28 [(270.3±73.1),(323.9±72.7),(758.7±245.9),(1020.6±383.6) ng/g,all P < 0.05]. Concentration of iAs,DMA of liver in 10,50,100 mg/L groups were higher than that of 0 mg/L group [(1.4±3.5),(49.7± 47.1),0,(100.4±30.2)ng/g]on PND 28,42 [iAs: (37.5±28.1),(268.8±246.4),(307.2±339.9),(15.4±9.4),(479.1±161.1),(408.4±51.9)ng/g;DMA: (594.5±148.8),(3181.9±519.0),(4834.2±2568.4),(1061.8± 85.2),(3697.1±553.7),(4120.0±732.8) ng/g,all P < 0.05]. Concentration of DMA of liver in 10,50,100 mg/L groups were higher than that of 0 mg/L group[(13.2±20.5)ng/g]on PND 15[(182.0±60,2),(637.6±90.0),(1458.7±196.3)ng/g,all P < 0.05]. Concentration of arsenicals of liver and brain showed a dose-dependent increase. The concentrations of DMA of breast milk in 50,100 mg/L groups were also higher than that of 0 mg/L group[(9.8±13.4),0 ng/g]on PND 0,15 [(182.3±85.9),(372.2±203.9),(124.2±33.1),(244.4±196.5)ng/g,all P < 0.05]. In the analysis of the change of arsenic on different postnatal day,we found the concentration of iAs,MMA,DMA,TMA in liver and brain of pups all decreased on postnatal day 15,and was lower than that on PND 0,28 and 42. Conclusions The distribution of arsenic and methyl-metabolism in liver and brain of pups is related with arsenic exposure dose. Arsenic can penetrate the placenta and blood brain barrier easily and lactation can hinder arsenic intake in some extent.

4.
Article in Chinese | WPRIM | ID: wpr-297072

ABSTRACT

<p><b>OBJECTIVE</b>To construct eukaryotic expression vector of the truncated septin2 and investigate the influence on the cultured mouse epidermal cell and fibroblast in vitro exerted by the transgenic expression product.</p><p><b>METHODS</b>The short splicing fragment was obtained by amplifying the reverse transcription product of the fetal mouse skin mRNA with PCR. Then its recombinant expression vector pcDNA3.1 (-)/septin2s was constructed and used to transfect the mouse epidermal cell and fibroblast cultured in vitro. The expression of the foreign gene was detected with RT-PCR and the changes of cell proliferation were observed and analysed.</p><p><b>RESULTS</b>RT-PCR results indicated that pcDNA3.1/septin2 was expressed in the cultured mouse epidermal cells and fibroblasts in vitro. We found that the epidermal cells accelerated their reproduction, but the fibroblasts had no obvious changes.</p><p><b>CONCLUSION</b>We successfully constructed eukaryotic expressive vectors of pcDNA3.1/ septin2s and transfected it into mouse epidermal cells and fibroblasts in vitro. The results settle a basis for showing effect of septin2s on fetal mouse skin.</p>


Subject(s)
Animals , Cell Line , Cytoskeletal Proteins , Genetics , DNA, Recombinant , Epithelial Cells , Cell Biology , Fibroblasts , Cell Biology , GTP-Binding Proteins , Genetics , Genetic Vectors , Mice , Mice, Inbred BALB C , Septins , Skin , Cell Biology , Transfection
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