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Objective To investigate the effect of SABP, a water-soluble component of Salvia miltiorrhiza, on the growth of orthotopic transplantation of H22 liver cancer and the immune microenvironment of liver cancer. Methods We established a mouse model of orthotopic transplantation of H22 cell liver cancer in BALB/c mice. ELISA was used to detect the expression of PD-L1, TGF-β, IL-1β, IL-10, IL-4, IFN-γ, IL-18, IL-7, IL-2, CCL-2 and CCL-21 in the liver. We counted the organ indexes of liver, spleen and kidney. Results SABP inhibited the growth of orthotopic transplantation tumors of H22 cell liver cancer, and increased the expression levels of PD-L1, TGF-β, IL-1β and IL-10 in the microenvironment of liver cancer, as well as the liver, spleen and kidney coefficients. Conclusion SABP could inhibit the growth of orthotopic transplantation tumors of H22 cell liver cancer and promote the expression of PD-L1, TGF-β, IL-1β and IL-10 in the microenvironment of liver cancer.
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Objective To investigate the effect of medical sodium hyaluronate gel (HA) on the growth and metastasis of abdominal and pelvic tumor cells in vitro and in nude mice. Methods Three tumor cells, Hela, CT26 and HCT116, were used to investigate the effects of different HA concentrations on the growth and migration of tumor cells in vitro by MTT assay and Transwell assay. An orthotopic transplantation model of colonic tumor in nude mice was established to investigate the effect on the proliferation of cell HCT116 by comparing the tumor volume and tumor mass 4 weeks after inoculation. The effects on the metastasis of cell CT26 were investigated by comparing the tumor metastasis rate and the number of metastatic lesions of lung and liver in nude mice among the different experimental groups 3 weeks after inoculation. Results HA did not promote the growth and metastasis of Hela, CT26 and HCT116 cells in vitro at different concentrations. Actually, HA exhibited a certain inhibitory activity at the concentration of 5 mg/ml. In the orthotopic transplantation model of colonic tumor-HCT116, HA did not promote the growth of cell HCT116. In the orthotopic transplantation model of colonic tumor-CT26, HA inhibited CT26 tumor metastasis. Conclusion Under the experimental conditions, HA did not promote the growth, migration or metastasis of abdominal and pelvic related tumor cells including Hela, CT26 and HCT116 in vitro and in vivo.
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Objective: To investigate the effect of inhibiting CIP2A expression on the biological characteristics of HepG2 cells. Methods: The recombinant adeno-associated viral vector rAAV2-EGFP-CIP2A shRNA was successfully transfected into HepG2 cell lines by using previous experiments. Cell proliferation was detected by MTT assay, cell apoptosis was measured by flow cytometry, and cell invasion ability was detected by Transwell assay. A model of subcutaneous implant tumor in nude mice was established to observe the growth of xenograft tumor in nude mice. The volume and quality of tumor growth were detected in nude mice after 30 days, and the expression of CIP2A protein in the transplanted tumor tissues was detected by immunohistochemistry. Results: The results of MTT and flow cytometry showed that the proliferation and apoptosis of HepG2 cells were inhibited after downregulation of CIP2A expression. The Transwell invasion experiment indicated a decrease in cell invasion ability. The result of subcutaneously growing tumor in the nude mice found in the interference experiment group of nude mice, the subcutaneous tumor growth rate, the final volume and weight were much smaller than those in blank control group and no-load virus group, CIP2A protein expression in transplanted tumor tissues was significantly lower. Conclusion: CIP2A promotes the proliferation, invasion of HepG2 cells and malignant growth in vivo.
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In this paper,the effects of active fractions of Ferula ferulaeoides on the growth and apoptosis of human gastric cancer cell MGC-803 transplantation tumor were systematically studied. The subcutaneous ectopic transplantation tumor model was established in human gastric cancer MGC-803 nude mice by cell suspension implantation method. The anti-tumor rate and organ index were used to evaluate the anti-tumor effect of the active fractions of F. ferulaeoides on the tumor-bearing nude mice. HE staining,TUNEL staining,RT-PCR,Western-blot and ELISA were used for pathological examination,apoptosis observation,and detection of apoptosis-related genes,proteins and cytokines expression. The results showed that as compared with the model group,the low,medium and high doses of the active fraction of F. ferulaeoides had inhibitory effects on xenografts in nude mice,respectively,in a dose-dependent manner; the apoptotic ratio was increased with the increase of drug concentration. As compared with the model group,F. ferulaeoides could down-regulate the expression of survivin mRNA in nude mice,and the protein expression levels of Bax,Bcl-2,caspase-3 and caspase-9 in tumor tissues of nude mice could be increased to different degrees in F. ferulaeoides groups. The contents of IL-10 and TGF-β1 in plasma of nude mice were decreased in high dose group of F. ferulaeoides active fractions. The results indicated that F. ferulaeoides can significantly inhibit the growth of human gastric cancer MGC-803 subcutaneously transplanted tumor,and its mechanism may be related with down-regulating the expression of survivin mRNA,and up-regulating the expression of apoptosis-related proteins Bax,caspase-3 and caspase-9.
Subject(s)
Animals , Humans , Mice , Apoptosis , Caspase 3 , Metabolism , Cell Line, Tumor , Cytokines , Metabolism , Ferula , Chemistry , Mice, Inbred BALB C , Mice, Nude , Neoplasm Transplantation , Plant Extracts , Pharmacology , Proto-Oncogene Proteins c-bcl-2 , Metabolism , Stomach Neoplasms , Drug Therapy , bcl-2-Associated X Protein , MetabolismABSTRACT
Objective: To investigate the effect of Buzhong Yiqi Tang on cisplatin resistance tumor cells A549/DDP in nude mice with lung adenocarcinoma, and determine the optimal concentration. Method: The 36 BALB/C nude mice were randomly divided into no-load group, model group, cisplatin group, low-dose Buzhong Yiqi Tang group (1.46 g · kg-1), medium-dose Buzhong Yiqi Tang group (2.92 g · kg-1) and high-dose Buzhong Yiqi Tang group (5.84 g · kg-1). The no-load group was inoculated with A549/DDP cells transfected by uninfected lentiviral transforming growth factor-β1(TGF-β1), and the other groups were inoculated with TGF-β1 over-expression A549/DDP cells. After the intervention, the nude mice were put to death. The tumors of mice were weighed, and the inhibition rate was calculated, liver, spleen and lung metastases of tumors were counted under microscope, and pathological observation was made for lung histomorphological changes. Immunohistochemistry, Western blot and Real-time polymerase chain reaction(Real-time PCR) were used to detect the expressions of lung resistance-related protein (LRP), multidrug resistance-related protein (MRP), P-glycoprotein (P-gp) in transplantation tumor. Result: Compared with the model group, the weight of tumors and the number of pulmonary metastatic nodules decreased in the Buzhong Yiqi Tang intervention group. With the increase of the dosage of Buzhong Yiqi Tang, the weight of tumors gradually decreased, while the inhibition rate increased. The effect of the high-dose group was the most significant (PPPConclusion: Buzhong Yiqi Tang can inhibit the growth and metastasis of transplanted tumors, alleviate lung injury and improve the drug resistance of A549/DDP to cisplatin, with a certain dose-effect relationship. High-dose concentration (5.84 g ·kg-1) was the best concentration in nude mice.
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Objective To investigate the inhibitory effects of high mobility group chromosomal protein N2 (HMGN2)on growth of human bladder cancer T24 cells and ectopic tumor growth of nude mice. Methods MTT and flow cytometry assay were conducted to detect cell growth of bladder epithelial cells(T24)cells in vitro. The transplantation tumor models in nude mice were constructed by injecting T24 cells in vivo. The para-tumorswere injected with PBS,HMGN2 protein and cisdichlorodiamineplatinum(DDP),respectively. Tumor volume and weight were calculated. The expression of cell proliferation-related proteins was detected by Western blot assay. Results MTT assay proved that HMGN2 could significantly inhibit the growth of T24 cells. Flow cytometry assay verified that HMGN2 could block T24 cells in S stage of the cell cycle. The average tumor volume and weight in the HMGN2 group and DDP positive control group were smaller than those in the PBS group(P<0.05,respectively), with the tumor inhibitory rate of 25% and 23%,respectively. The results of Westernblot showed that HMGN2 could decrease Bcl-2 expression and increase Bax expression in tumor. Conclusion HMGN2 has a significant antitumor effect on T24 cells and bladder carcinoma in nude mice,which may be associated with the induction of the apoptosis of carcinoma cells and the regulation of the cell cycle.
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@#The study aims to establish a human gastric cancer orthotopic transplantation model in nude mice and to use 7T MRI for detection. After poorly differentiated human gastric adenocarcinoma SGC-7901 cells were injected subcutaneosly into the right flanks of nude mice, the model of in nude mice was established with orthotopic transplanted cancer of gastric tumor by the Compont® gel pasted method. 7T MRI scan was conducted on the mice after operating model about 20 days later. Histopathological examinations were carried out on the stomach. Two of three mice on which 7T MRI scan were performed showed visible suspected stomach tumor and their presence was verified again by histopathological examinations; tumor formation rate in the nude mice gastric orthotopic transplantation model was 66. 7%. This study suggested that 7T MRI could be used in the live detection of in situ tumor and that MRI could be used for pre-clinical gastric cancer drug development and clinical gastric carcinoma diagnosis.
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OBJECTIVE:To study the inhibitory effects of sinomenine(SIN)-loaded polylactic-co-glycolic acid-D-α-tocopherol polyethylene glycol 1000 succinate(PLGA-TPGS)nanoparticles(SPTN)on the proliferation of HCa-F cells in lymph tubes and ec-topic transplantation tumors in mice. METHODS:HCa-F cell suspension were incubated with normal saline,5-fluorouracil(FS), sinomenine solutions (SS),sinomenine PLGA nanoparticles (SPN) and SPTN,with concentration of 80 μg/ml. The cells were marked with CFSE,and then were injected with suspension 50 μl via one footpad of mice. Inhibitory effect of above suspensions on the proliferation of HCa-F in lymph tubes of mice was observed by fluorescence inverted microscope at 3,6,9,12,24 h(n=15). Mice were divided into normal control group,blank PLGA-TPGS nanoparticles (EPTN) group,normal saline group,SPTN group,SPN group,SS group and FS group with 10 mice in each group. The latter 5 groups were injected with relevant medicine 15 mg/kg,once a day,via tail vein for consecutive 10 days after the model of HCa-F cells-bearing ectopic transplantation tumor mice was established. The serum content of ALT,AST,γ-glutamyltransferase(γ-GT),ALB and T-BIL were determined;solid tu-mors were taken,measured and weighed,and the inhibitory rate of tumor was also calculated. RESULTS:The inhibitory effect of above solution on the proliferation of HCa-F cells in descending order was as follows:SPTN>SPN>FS>SS>normal saline. Com-pared with normal saline group,the serum levels of ALT,AST,γ-GT and TBIL of SPTN group,SPN group,SS group and FS group decreased,while ALB level increased(P<0.05);the amount of tumor volume increase and tumor weight in SPTN group, SPN group and FS group decreased significantly (P<0.05). The inhibitory rate of tumor in 3 groups were 49.62%,40.53% and 33.90%. CONCLUSIONS:SPTN can inhibit the proliferation of HCa-F cells in lymph tubes of mice,and can improve HCa-F cells-bearing ectopic transplantation tumor in mice. It is better than SPN and FS.
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Objective To observe the influencing of Yiqi Huayu Jiedu Prescription on the growth of HepG2 nude mice transplantation tumor and the expression of related factors of vascular mimicry. Methods Models of transplanted tumors, which were made by HepG2 cells in nude mice, were randomly divided into 7 groups, Yiqi Huayu Jiedu Prescription group, Astragali Radix group, Curcumae Rhizoma group, Paridis Rhizoma group, Gecko group, cis-platinum group, and model group. Except for the model group, the rest groups were given relevant medicine for intervention. 21 days later. HIF-1α, MMP-2, MMP-9, and E-cad were detected by immunohistochemistry, and Twist1 and Bcl-2 were detected by fluorescence quantitative PCR. Results Compared with the model group, tumor volume in the rest groups decreased (P<0.05), and the effect in the Yiqi Huayu Jiedu Prescription group was more obvious than the Astragali Radix group, Paridis Rhizoma group and Gecko group (P<0.05);The expression of vasculogenic mimicry structure was rare in each group, and the model group and cis-platinum group were the most obvious;Except for the Astragali Radix group, the expressions of HIF-1α, MMP-2, and MMP-9 showed statistical significance compared with model group (P<0.05);The expression of E-cad in the Yiqi Huayu Jiedu Prescription group and Astragali Radix group showed statistical significance (P<0.05);The expression of Bcl-2 in the Yiqi Huayu Jiedu prescription group, Paridis Rhizoma group, and Gecko group decreased significantly compared with the model group (P<0.05);The expression of Bcl-2 in the Yiqi Huayu Jiedu prescription group was much better than the other groups (P<0.05);The expression of Twist1 showed statistical significance in the Yiqi Huayu Jiedu Prescription group, Curcumae Rhizoma group, Paridis Rhizoma group, and cis-platinum group (P<0.05). Conclusion Yiqi Huayu Jiedu Prescription can reduce expression of HIF-1α, Twist1, Bcl-2, MMP-2, and MMP-9, and increase expression of E-cad, thereby inhibiting the formation of vascular mimicry.
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Objective To investigate the characters and targeted ability of FITC-CSNRDARRC molecular probe in labeling orthotopic transplantation tumor of bladder cancer in vivo.Methods From July 2013 to June 2014,the stability and characters of FITC-CSNRDARRC molecular probe were detected by spectrophotometer and molecular imaging and the optimum concentration and imaging time window were determined.30 BALB-C nude mice were randomly divided into experimental group (n =20) and control group(n =10).In control group,5 of them (group A) were ligated bilateral ureter,others(group B) were not.We established orthotropic transplanted bladder tumor (BIU-87) model by operation.And 0.2 ml probes (220 μmol/L) was then injected intravenously in all mice after 2 weeks.We obtained images and analyzed average gray value of the heart,lung,liver,spleen,bilateral kidney and orthotropic transplantation bladder tumor by using optical probe molecule fluorescence imaging system after 30 min,1 h,2 h,4 h and 12 h,respectively.Results After injected the FITC-CSNRDARRC molecular probes intravenously at 220 μmol/L,the fluorescence signal of tumor tissue strengthened gradually.The optimal imaging time window was 4 hours after injection.The illumination and temperature had little effect on the fluorescence signal.With the time passing after injection,the intensity of florescence signal progressively increasing,which reached the peak at4 h.The average gray value of tumor tissue at 1 h,2 h,3 h,4 h,5 h,6 h,8 h,12 h were 74.22,76.2,80.11,89.38,83.29,85.1,81.22,83.01,respectively.The fluorescence signal of normal tissue weakened gradually with the passage of time.Only liver and gall bladder could notice the fluorescence signal 4 hours after injection in group A.However,the relatively strong fluorescence signal could be found in liver and gall bladder in group B.Conclusions The characters of fluorescence probe are affected by its concentration.Its optimal concentration of labeling tumor is 220 μmol/L.The optical imaging time window was about 4 h after intravenous injection.The FITC-CSNRDARRC molecular probe can specifically bound to orthotopic transplanted tumor of bladder cancer in vivo.
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Objective To investigate the mechanism of Kanglaite combined with 5-Fu in treating pancreatic carcinoma through studying the changes of Bcl-2,Bax and vascular endothelial growth factor(VEGF) in pancreatic carcinoma cell and the relationship between apoptosis and these changes.Methods The model of athymic mouse subcutaneous transplantation tumor of human pancreatic carcinoma PC-3 cell was established;the expression of Bcl-2,Bax and VEGF were detected in malignant cells using immunohistochemistry;the apoptosis proportion was detected by adopting flow cytometry;the tumor size was detected.Results Kanglaite combined with 5-Fu could obviously depress the expression of Bcl-2 in protein level in malignant cells;apoptosis proportion had no effect on Bax expression;Kanglaite could obviously depress the expression of VEGF in malignant cells.The apoptosis proportion was higher and protein expression of Bcl-2 was obviously lower in group of Kanglaite combined with 5-Fu than that in Kanglaite group or 5-Fu group.What was more,the protein expression of VEGF and the tumor volum was lowest in group of Kanglaite combined with 5-Fu.Conclusion The effect of Kanglaite united with 5-Fu on treatment of pancreatic cancer is better than that of Kanglaite or 5-Fu alone.
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OBJECTIVE:To study the inhibitory action of Sarcandra glabra injection (ZJF) on tumor growth of SGC-7901 transplantation tumor in nude mice and its induction on apoptosis. METHODS:SGC-7901 transplantation tumor nude mice model was established then the model mice were assigned to blank,positive control,ZJF high-dose and low-dose (3,1.5 g?kg-1) groups. After treatment with ZJF for 14 days,the indexes of thymus and spleen,numbers of peripheral red blood cells,white cells and platelet were measured; the tumor-inhibition rate was computed; the ultrastructure of tumor tissues was observed,and the apoptosis of implanted tumor cells was observed. RESULTS:As compared with blank group,no significant change was noted in the indexes of thymus and spleen and peripheral hemogram. The differences between high-dose ZJF group and low-dose ZJF group were greatly significant in both the inhibition rate of SGC-7901 transplantation tumor (42.8% vs. 39.9%) and the apoptotic rate of tumor tissues [(9.3?0.15)% vs.(8.18?0.08)%].One or more apoptotic bodies formed after medication. CONCLUSION:ZJF can promote the presence of apoptotic body and induce apoptosis thus inhibiting the growth of SGC-7901 transplantation tumor in nude mice; however,it had little impact on the thymus and spleen and peripheral hemogram.
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Objective: To observe the morphological characteristics of lymphatics and to investigate the intratumoral lymphangiogenesis in transplantation tumor model of human breast infiltrating duct carcinoma in nude mice.Methods: Small slices of human breast carcinoma tissue were transplanted into 5 nude mice to establish implanted tumor model.HE staining was used to observe the characteristics of the transplantation tumors.Vascular endothelial growth factor-3(VEGFR-3) label staining was performed to observe lymphatics,and ultrastructure of lymphatics was observed with transmission electron microscope(TEM).Results: Tumors were viable in a11 the mice,and grew up to about 0.6~0.8 centimeters in diameter after 52 days when specimen was collected.Under light microscope,lymphatics were found in all the tumors,and were located seemingly much more in periphery than in the center of the tumors.Under TEM some characteristics of neogentic lymphatics could be seen,such as thick wall,narrow lumen,changes of organell,etc.Conclusion: Neogentic lymphatics can be seen in transplantation tumor model of human breast infiltrating duct carcinoma in nude mice.