Effect of Xiangsha Liu Junzitang Combined with Phlegm-removing and Detoxifying Traditional Chinese Medicine on Immune Escape in Lewis Lung Cancer Mice / 中国实验方剂学杂志

ObjectiveTo study the effect and mechanism of Xiangsha Liu Junzitang combined with phlegm-removing and detoxifying traditional Chinese medicine on immune escape in Lewis lung cancer mice. MethodA total of 60 specific-pathogen-free （SPF）-grade C57BL/6J male mice were injected subcutaneously with 0.2 mL of Lewis cell suspension （containing 2×106 cells·mL-1） in the right mid-axillary line. After 7 days， the mice that had been successfully modeled were randomly divided into six groups： the model group， the cisplatin group， the Xiangsha Liu Junzitang low-， medium-， and high-dose groups， and the combined group， with 10 mice in each group. The Xiangsha Liu Junzitang low-， medium- and high-dose groups were gavaged with 17.88， 35.75， 71.50 g·kg-1 Xiangsha Liu Junzitang solution once a day， respectively， and the dosage of cisplatin intraperitoneally injected into the mice was converted to 5 mg·kg-1 twice a week， and the tumour volumes of each group were measured every two days. The intervention lasted for 14 consecutive days. At the end of treatment， the tumour mass of mice in each group was weighed and the tumour inhibition rate was calculated. The morphological characteristics of tumours in each group were observed by hematoxylin-eosin （HE） staining. Fluorescent quantitative real-time polymerase chain reaction （Real-time PCR） assay was used to detect messenger ribonucleic acid （mRNA） contents of the natural killer group 2 member D （NKG2D） receptor， ribonucleic acid export-1 （RAE-1）， and γ interferon （IFN-γ） in the tumour tissues of each group. NKG2D， RAE-1， and IFN-γ mRNA in tumour tissues of each group. Immunohistochemistry （IHC） and Western blot were applied to detect the expressions of RAE-1， NKG2D， and IFN-γ in tumour tissues of each group， and Western blot was used to detect the expressions of interleukin-6 （IL-6）， Janus kinase 2 （JAK2）， p-JAK2， signal transducer and activator of transcription 3 （STAT3）， and p-STAT3 in tumour tissues of each group， as well as the protein levels of NKG2D， and RAE-1 in spleen tissues of each group. ResultCompared with that in the model group， the tumour mass decreased in all dose groups of Xiangsha Liu Junzitang， with no statistically significant difference. The tumour volume was reduced （P<0.05， P <0.01）. The pathological morphology was improved. The mRNA contents of NKG2D， RAE-1 and IFN-γ were increased in the medium-dose group （P<0.05， P<0.01）， and the protein expressions of NKG2D， RAE-1， and IFN-γ in tumour tissues were elevated （P<0.05， P<0.01）， and p-JAK2 and p-STAT3 protein expressions were decreased （P<0.05， P<0.01）. In spleen tissues， the protein expressions of NKG2D and RAE-1 in all dose groups of Xiangsha Liu Junzitang were significantly elevated （P<0.01）. Compared with those in the cisplatin group， NKG2D， RAE-1 and IFN-γ mRNA contents were elevated in the middle-dose group of Xiangsha Liu Junzitang， and the difference was not statistically significant. IHC showed that the protein expressions of NKG2D and IFN-γ in the combined group were significantly elevated （P<0.01）， and Western blot results showed that the protein expressions of RAE-1， NKG2D and IFN-γ were elevated （P<0.05， P<0.01）. p-JAK2 and p-STAT3 protein expressions were decreased in the combined group （P<0.05， P<0.01）. NKG2D and RAE-1 protein expressions were significantly increased in spleen tissues of the medium-dose groups and the combined group （P<0.01）. ConclusionXiangsha Liu Junzitang combined with phlegm-removing and detoxifying traditional Chinese medicine can inhibit the growth of tumours in Lewis lung cancer mice by up-regulating the expressions of RAE-1/NKG2D， promoting the activation of NK cells， and inhibiting immune escape， the mechanism of which may be related to down-regulation of the JAK2/STAT3 pathway.

Therapeutic effect of Xiakucao Xiaoliu mixture on Lewis lung cancer mice / 药学实践杂志

Objective To investigate the therapeutic effect of Xiakucao Xiaoliu mixture on Lewis lung cancer mice. Methods 30 mice with C57BL/6 mouse Lewis lung cancer xenograft model were randomly divided into three groups: model control group, Xiakucao Xiaoliu mixture group (M group), cisplatin group (DDP group). M group and DDP group were administered continuously for 14 days. Through the general observation of Lewis lung cancer mice, tumor size was determined, HE staining method was used to determine the histopathological changes of tumors, and the expression of CyclinD1 and P16 in tumor tissues was determined by immunohistochemistry. Results The tumor weight of the model control group was the heaviest, and the difference was statistically significant compared with other groups. (P<0.05). Survival state and quality of life of mice had been improved to some extent in M group. The results of tumor growth curve and HE staining in each group of mice showed that the growth of tumor cells had been inhibited and normal cells had been protected. The positive expression of CyclinD1 was significantly decreased in M group and DDP group (P<0.01), but the effect of M group on the improvement of P16 positive expression was not significant. Conclusion Xiakucao Xiaoliu mixture had a good effect on inhibiting lung tumor growth.

Study on Inhibitory Effect of Modified Maimendong Decoction Combined with Cisplatin on Lewis Lung Cancer Transplantation Model Mice and Its Mechanism / 中国药房

OBJECTIVE： To study the inhibitory effect of modified Maimendong decoction combined with cisplatin on Lewis lung cancer transplantation model mice， and to explore its potential mechanism. METHODS： Lewis lung cancer transplantation mice model was induced via subaxillary inoculation of Lewis lung cancer cells. 60 mice were randomly divided into model group （normal saline， once a day， i.g.）， cisplatin group （6 mg/kg， once a week， i.p.）， modified Maimendong decoction group （20 g/kg. once a day， i.g.） and combination group （cisplatin 6 mg/kg， once a week， i.p.+modified Maimengdong decoction， once a day， i.g.）， with 15 mice in each group. All mice were treated for consecutive 2 weeks. After medication， tumor weight and thymus index were detected； HE staining was used to observe the pathological change of tumor tissue. TUNEL was used to detect apoptotic index of tumor tissue. The protein expressions of Bcl-2 and Bax were detected by Western blot assay. RESULTS： Compared with model group， tumor weight and protein expression of Bcl-2 were decreased significantly in modified Maimengdong decoction， cisplatin group and combination group （P＜0.05）， and thymus index， tumor apoptotic index and the protein expression of Bax were increased significantly （P＜0.05）. Tumor weight and protein expression of Bcl-2 in combination group were significantly lower than modified Maimengdong decoction group and cisplatin group （P＜0.05）； thymus index， tumor apoptotic index and the protein expression of Bax were significantly higher than addition and subtraction of modified Maimengdong decoction group and cisplatin group （P＜0.05）. HE staining showed that the density of tumor cells was decreased to certain degree in modified Maimengdong decoction group， cisplatin group and combination group； the area of necrosis area in the combination group was significantly larger than in modified Maimendong decoction group and cisplatin group. CONCLUSIONS： Modified Maimendong decoction can inhibit the growth of Lewis lung cancer transplanted tumor in mice by down-regulating the protein expression of Bcl-2 and up-regulating the protein expression of Bax.

Inhibition of Total Flavonids from Cudrania tricuspidata Bun on Lewis Lung Cancer and the Effect of Its Component Compatibility on Autophagy of LLC Cells / 中国药学杂志

OBJECTIVE: To explorethe effect of total flavonoids from Cudrania tricuspidata Bun on Lewis lung cancer mice and its new component compatibility on the autophagy of LLC cells. METHODS: A mouse model of Lewis lung cancer xenografts was constructed. The body weight, tumor weight, tumor volume and organ index were measured before and after taking total flavonoids of Cudrania tricuspidata Bun. The HE staining of the xenograft pathological sections were also observed. The TNF-α, IL-2, IL-6 and IL-12 levels in the serum were calculated by ELISA. The content of the main active ingredient and its ratio in the flavonoid extract were measured by UPLC. Western blot was used to detect the effect of the new component compatibility on the expression of autophagy protein in LLC cells, and the ultra structural changes of LLC cells were observed by transmission electron microscopy. Flow cytometry was used to detect the effect of the new component compatibility on autophagy of LLC cells. RESULTS: The high-dose group of total flavonoids from Cudrania tricuspidata Bun can significantly inhibit the growth of tumor in mice and enhance the organ index of tumor-bearing mice, and the survival rate of mice could be improved in all groups of total flavonoids from Cudrania tricuspidata Bun. The serum levels of TNF-α, IL-2, IL-6 and IL-12 in the tumor-bearing mice of each group were higher than those in the model group, and there was significant difference in the high dose group of total flavonoids (P<0.01). The main active ingredients were taxifolin, kaempferol and naringenin by UPLC. The ratio of taxifolin to kaempferol was 60:1. Western blot assay showed that its new component compatibility significantly increased the expression of autophagic protein in LLC cells (P<0.01). The autophagosomes in the cytoplasm were observed by transmission electron microscopy. The results of flow experiments showed that the average fluorescence intensity of its new component compatibility was significantly higher than that of the control group. CONCLUSION: Total flavonoids of Cudrania tricuspidata Bun can effectively inhibit tumor growth and have less harm to immune organs. Its mechanism may be related to up-regulation of cytokines TNF-α, IL-2, IL-6 and IL-12 levels and improve immune function in the body. Moreover, its main active ingredient promotes the increase of autophagy protein expression in LLC cells and induces autophagy in LLC cells.

Preparation of docetaxel-loaded nanomicelles and their anti-Lewis lung cancer effect in vitro / 中国中药杂志

Docetaxel-loaded nanomicelles were prepared in this study to improve the solubility and tumor targeting effect of docetaxel(DTX),and further evaluate their anticancer effects in vitro. PBAE-DTX nanomicelles were prepared by film-hydration method with amphiphilic block copolymer polyethyleneglycol methoxy-polylactide(PELA) and pH sensitive triblock copolymer polyethyleneglycol methoxy-polylactide-poly-β-aminoester(PBAE) were used respectively to prepare PELA-DTX nanomicelles and PBAE-DTX nanomicelles. The nanomicelles were characterized by physicochemical properties and the activity of mice Lewis lung cancer cells was studied. The results of particle size measurement showed that the blank micelles and drug-loaded micelles had similar particle sizes, ranging from 10 to 100 nm. The particle size of PBAE micelles was changed under weak acidic conditions, with good pH response. The encapsulation efficiency of the above two types of DTX-loaded nanomicelles determined by HPLC was(93.8±1.70)% and(87.2±4.10)%, and the drug loading amount was(5.3±0.10)% and(4.9±0.05)%,respectively. Furthermore,the DTX micelles also showed significant inhibitory effects on Lewis lung cancer cells by MTT assay, and pH-sensitive PBAE-DTX showed better cytotoxicity. The results of flow cytometry indicated that,the apoptosis rate of lung cancer Lewis cells was(20.72±1.47)%,(29.71±2.38)%,and(40.91±1.90)%(P<0.05) at 48 h after treatment in DTX,PELA-DTX,and PBAE-DTX groups. The results showed that different docetaxel preparations could promote the apoptosis of Lewis cells, and PBAE-DTX had stronger apoptotic-promoting effect. The pH-sensitive DTX-loaded micelles are promising candidates in developing stimuli triggered drug delivery systems in acidic tumor micro-environments with improved inhibitory effects of tumor growth on Lewis lung cancer.

Effect of Maimendong Tang and Qianjin Weijingtang on Expression Profiles of lncRNA and mRNA in Mouse Orthotopic Lewis Lung Carcinoma / 中国实验方剂学杂志

Objective: To explore the anti-lung cancer mechanism of Maimendong Tang and Qianjin Weijingtang (Jin Fang) by detecting the expression profiles of long noncoding RNA (lncRNA) and mRNA in mice tumor tissues of orthotopic Lewis lung cancer model. Method: C57BL/6 mice were randomly divided into normal group, model group and Jin Fang group (20 g·kg-1·d-1). After successful establishment of Lewis lung cancer model in situ in mice, Jin Fang was given orally the next day after treatment. Using gene chip technology, differential lncRNA and mRNA closely related with Jin Fang' s anti-lung cancer effect were detected, and cluster analysis was performed. The key lncRNA and mRNA were screened out by t-test and fold change of differential expression. Bioinformatic methods were used to predict target genes regulated by differential lncRNA, and functional and pathway analysis was performed. The histopathological technique was used to detect the differences in the tumor tissue of each group under light microscope. Result: lncRNA and mRNA chip hybridization results showed that Jin Fang regulated differential expressions of 887 lncRNA, in which 442 were up-regulated and 445 were down-regulated (PPConclusion: Jin Fang may exert its anti-lung cancer effect by regulating the expressions of multiple lncRNAs and mRNAs, and down-regulating related signaling pathways.

Evaluation of cytotoxicity of stearoyl gemcitabine-loaded PLGA nanoparticles to Lewis lung cancer cells / 中国免疫学杂志

Objective:To investigate the anti-proliferation effect of 4-(N)-stearoyl gemcitabine-loaded poly(lactic-co-glycolic) acid nanoparticles(GemC18-PLGA-NPs) on Lewis lung cancer cells(LLC) in vitro.Methods: Lewis cells were incubated with GemC18-PLGA-NPs,free GemC18,gemcitabine HCl(GemHCl) or GemC18-free blank nanoparticles(PLGA-NPs) respectively and cell viability was determined using an MTT assay after 24,48 or 72 h of incubation.The apoptosis rate after 48 and 72 h of incubation were measured by flow cytometry.Results:GemC18-PLGA-NPs,GemC18,and GemHCl all significantly inhibited the growth of LLC cells, and the survival rate of GemHCl group was lowest,GemC18-PLGA-NPs group had the highest survival rate.The cell survival rate of GemC18-PLGA-NPs after 72 h was significantly higher than that of GemHCl (P<0.05) at the concentration of 1 μmol/L,indicating that it had a significant drug release effect.PLGA-NPs group produced trifle inhibition on the Lewis cells without correlation to time or concentration.Conclusion:GemC18-PLGA-NPs have significant anti-proliferation effect on mouse Lewis lung cancer cells in vitro.

Estabilshment and stemness identification of an asymmetric dividing cell line derived from mouse Lewis lung cancer cells / 基础医学与临床

Objective To establish an asymmetric dividing cell line(LLC-ASD cells) derived from mouse Lewis lung carcinoma cancer cells(LLC-parental cells),and to investigate its stemness features in order to lay a founda-tion for depth studying the function of asymmetric dividing in the cancer biology. Methods In order to obtain asymmetrically dividing LLC cells (LLC-ASD cells) derived from LLC-Parental cells,8 times of consecutive cul-ture,enrichment and collection of floating spheriod forming cells followed by 5 times of consecutive single cell clo-ning were conducted. Immunofluorescence assay was used to visualize and quantify the rate of asymmetric division in LLC-ASD cells labeled by BrdU. For comparing the stem characteristics of LLC-Parental and LLC-ASD, RT-qPCR,clonogenic assay in 6-well plate,single cell spheroid formation assay with agar in 6-well plate and 96-well-plate single cell cloning assay were conducted. In vivo,LLC-parental cells and LLC-ASD cells were subcutaneously transplanted in nude mice to determine the effect of the difference in stem cell like properties on tumorigeneicy. The same lung transplantation into tumor experiment in mice were used to compare the differences in cancer biology. Results Asymmetric dividing cells were found in LLC-ASD cell culture through the BrdU immunofluorescence assay and the rate of asymmetric division in the anaphase cells was as high as 50%。According to the clonogenic assay in 6-well plate,single cell and spheroid formation assay with agar in 6-well plate and 96-well-plate single cell cloning assay in LLC-ASD cells,the results showed that they were more prominent than those in the LLC-Parental cells(P<0.05). In vivo,the tumor metastasis potentials of LLC-ASD was enhanced than that of LLC-parental when transplanted to the C57 mice. Further,the tumorigenic potentials of LLC-ASD cells was also increased.Conclusions The asymmetric dividing cell line derived from mouse Lewis lung carcinoma cancer cell line (LLC-ASD cells) is established which exhibits stemness properties. The establishment and characterization of this model will facilitate the research on the function of asymmetric cell dividing in cancer biology.

Effects of total flavonoids from Cycas Revolute on expression of VEGF,bFGF,HIF-1α and NF-κB in model mice of Lewis lung cancer / 中国免疫学杂志

Objective:To investigate the effects of total flavonoids from Cycas Revolute on expression of vascular endothelial growth factor (VEGF),basic fibroblast growth factor (bFGF),Hypoxia inducible factor-1α (HIF-1α) and nuclear factor-κB(NF-κB) in model mice of Lewis lung cancer.Methods: The expressions of VEGF,bFGF,HIF-1α and NF-κB in tumor tissues were detected by immunohistochemistry and Western blot.The expression of VEGF,VEGF and NF-κB in tumor tissues were detected by fluorescent quantitative PCR.BFGF,HIF-1α and NF-κB mRNA were detected by immunohistochemistry.Results: The results of immunohis-tochemistry,Western blot and Real-time PCR showed that the results were basically the same,compared with model group,the expression of VEGF,bFGF,HIF-1α,NF-κB mRNA and the expression of VEGF,bFGF,HIF-1α and NF-κB were decreased,the difference was highly significant (P<0.05).Conclusion: The mechanism of total flavonoids from Cycas Revolute in the treatment of lung cancer may be through inhibition of the expression of VEGF,bFGF,HIF-1α,NF-κB in invasion and metastasis,and further inhibit the expression of VEGF,bFGF,HIF-1α and NF-κB in invasion and metastasis-related proteins,thus play a role of anti-lung cancer invasion and metastasis.

Effects of Shanxian Granule on antagonistic immunity of Lewis lung cancer mice and level of IFN-γ,TNF-β,IL-10 in peripheral blood / 中国免疫学杂志

Objective:To investigate the effects of Shanxian granule on proliferation of Lewis lung cancer cells and anti-tumor immunity and immune microenvironment of Lewis lung cancer-bearing mice in order to explore the molecular mechanism of anti-tumor of Shanxian Granule and improve the anti-tumor immunity of the body, and provide further theoretical basis for its clinical application. Methods:Lewis lung cancer cells was transplanted to axillary skin to establish mouse tumor model. The mice divided into blank group,model group,chemotherapy group and Shanxian granule group. The tumor tissue of Lewis lung cancer tumor bearing mice was weighed and the tumor inhibition rate was calculated. Immunohistochemical method was used to detect the expression of CD and CD8 in spleen tissue. The effect of lymphocytes on the proliferation of Lewis lung cancer cells was detected by CCK-8 method. The level of IFN-γ,TNF-βand IL-10 in peripheral blood were detected by ELISA. Results:①The tumor inhibition rate of Lewis lung cancer was 45. 99% in Shanxian Granule group,which was significantly higher than that of chemotherapy group (P<0. 05).②The lymphocytes of mouse can inhibit the proliferation of Lewis lung cancer cells and have a positive correlation with lymphocyte concentration and duration of action. Moreover,CD4+ T cells,CD4+/CD8+ratio and lymphocyte inhibition rate of Lewis lung cancer cells in model group and chem-otherapy group were significantly lower than those in blank group (P<0. 05). Shanxian granule group was significantly higher than the model group and chemotherapy group ( P<0. 05 ) . However, there was no significant difference between Shanxian granule group and blank group(P>0. 05).③The levels of IFN-γand TNF-βin peripheral blood of model group and chemotherapy group were significantly lower than those in blank group,while IL-10 was significantly higher than that in blank group (P<0. 05). The levels of IFN-γand TNF-βin peripheral blood of mice in Shanxian granule group were significantly higher than those in model group and chemotherapy group, while IL-10 was significantly lower than that in model group and chemotherapy group (P<0. 05). There was no significant difference in IFN-γ,TNF-β and IL-10 in peripheral blood of mice between Shanxian granule group and blank group. Conclusion:Shanxian granule can significantly inhibit the growth of tumor tissue of Lewis lung cancer tumor bearing mice,increase the spleen index of mice,enhance the activity of T lymphocytes,upregulate IFN-γ and TNF-β in peripheral blood and decrease IL-I. These suggested that the anti-tumor effect of Shanxian granule may be achieved by regulating the content of CD4+ T lymphocyte,the ration of CD4+/CD8+ and Th1/Th2 ratio,in order to restore the immune steady function of tumor patients,improve the immune system and enhance the immune surveillance function.

Effects of Shanxian Granule on antagonistic immunity of Lewis lung cancer mice and level of IFN-γ,TNF-β,IL-10 in peripheral blood / 中国免疫学杂志

Objective:To investigate the effects of Shanxian granule on proliferation of Lewis lung cancer cells and anti-tumor immunity and immune microenvironment of Lewis lung cancer-bearing mice in order to explore the molecular mechanism of anti-tumor of Shanxian Granule and improve the anti-tumor immunity of the body, and provide further theoretical basis for its clinical application. Methods:Lewis lung cancer cells was transplanted to axillary skin to establish mouse tumor model. The mice divided into blank group,model group,chemotherapy group and Shanxian granule group. The tumor tissue of Lewis lung cancer tumor bearing mice was weighed and the tumor inhibition rate was calculated. Immunohistochemical method was used to detect the expression of CD and CD8 in spleen tissue. The effect of lymphocytes on the proliferation of Lewis lung cancer cells was detected by CCK-8 method. The level of IFN-γ,TNF-βand IL-10 in peripheral blood were detected by ELISA. Results:①The tumor inhibition rate of Lewis lung cancer was 45. 99% in Shanxian Granule group,which was significantly higher than that of chemotherapy group (P<0. 05).②The lymphocytes of mouse can inhibit the proliferation of Lewis lung cancer cells and have a positive correlation with lymphocyte concentration and duration of action. Moreover,CD4+ T cells,CD4+/CD8+ratio and lymphocyte inhibition rate of Lewis lung cancer cells in model group and chem-otherapy group were significantly lower than those in blank group (P<0. 05). Shanxian granule group was significantly higher than the model group and chemotherapy group ( P<0. 05 ) . However, there was no significant difference between Shanxian granule group and blank group(P>0. 05).③The levels of IFN-γand TNF-βin peripheral blood of model group and chemotherapy group were significantly lower than those in blank group,while IL-10 was significantly higher than that in blank group (P<0. 05). The levels of IFN-γand TNF-βin peripheral blood of mice in Shanxian granule group were significantly higher than those in model group and chemotherapy group, while IL-10 was significantly lower than that in model group and chemotherapy group (P<0. 05). There was no significant difference in IFN-γ,TNF-β and IL-10 in peripheral blood of mice between Shanxian granule group and blank group. Conclusion:Shanxian granule can significantly inhibit the growth of tumor tissue of Lewis lung cancer tumor bearing mice,increase the spleen index of mice,enhance the activity of T lymphocytes,upregulate IFN-γ and TNF-β in peripheral blood and decrease IL-I. These suggested that the anti-tumor effect of Shanxian granule may be achieved by regulating the content of CD4+ T lymphocyte,the ration of CD4+/CD8+ and Th1/Th2 ratio,in order to restore the immune steady function of tumor patients,improve the immune system and enhance the immune surveillance function.

Effects of NF-κB/p65 siRNA on Apoptosis and Bax Expression in Lung Tumour Cell Xenografts of Nude Mouse / 中国医科大学学报

Objective To investigate the effects of nuclear factor-kappaB(NF-κB)/p65 signaling transduction on the apoptosis and expressions of apoptosis-related genes Bax in nude mouse lung tumour cell xenografts. Methods The nude mice Lewis lung carcinoma cell xenograft model was established,and then the mice were intraperitoneally injected with NF-κB/p65 small interfering RNA(siRNA). The apoptosis of xenografted tumor cells in nude mice was detected by TUNEL assay. Expressions of Bax mRNA and protein were detected by RT-PCR and Western blotting. Results The result of TUNEL assay demonstrated that p65 siRNA evidently evoked cell apoptosis. Compared to the PBS treatment group or the normal control mice,both mRNA and protein expression levels of Bax in tumor xenografts were significantly up-regulated. There were significant differences among three groups(P<0.05). Conclusion NF-κB/p65 subunit may play an essential role in cell apoptosis of Lewis lung tumor.

Protective Effects of Danhong Injection on Myocardial Injury Induced by Doxorubicin in Lewis Lung Canc-er Mice / 中国药师

Objective:To study the protective effects of Danhong injection ( DH) on myocardial damage induced by doxorubicin ( DOX) in Lewis tumor bearing mice. Methods:The model of Lewis lung cancer in mice was established by underarm injecting tumor cells, and then randomly divided into four groups:the model control group, DOX group, DH group and DH+DOX group. After the experiment, myocardial and tumor tissue were separated from Lewis tumor bearing mice, and the excised tumors were weighted. The activities of lactate dehydrogenase ( LDH) , creatine kinase ( CK) , manganese superoxide dismutase ( SOD) , catalase ( CAT) and glu-tathione peroxidase ( GPx) , and the content of malondialdehyde ( MDA) were determined by a colorimetric method. Flow cytometry was used to determine the levels of apoptosis, reactive oxygen species (ROS) and mitochondrial membrane potential (△Ψm). Re-sults:Compared with that in the model control group, a significant decrease of tumor weight was shown in both DOX group and DH+DOX group (P<0. 01). DH had no significant influence on the anticancer function of DOX. The activity of LDH and CK, and the ap-optosis in myocardium cells significantly increased (P<0. 01). Compared with DOX group, the activities of LDH and CK, and the ap-optosis significantly decreased in DH+DOX group (P<0. 01). The activities of △Ψm, SOD, CAT and GPx significantly increased (P<0.05orP<0.01). ThecontentofMDAandROSgenerationbothdecreased(P<0.01).Conclusion:DHhasnosignificantin-fluence on the antitumor effect of DOX. The combination of DH and DOX shows cadioprotective effect on the myocardial damage through improving mitochondrial antioxidant defense capacity, ameliorating oxidative stress and maintaining △Ψm homeostasis.

Effect of Xiaoyan Decoction on bone marrow suppression of mice with Lewis lung cancer / 中草药

Objective: To study the effects and mechanism of Xiaoyan Decoction on bone marrow suppression in mice from aspects of peripheral blood cell ratio, immune function, and cytokines. Methods: A mouse model of bone marrow suppression was established. Fifty mice were randomly divided into five groups (n = 10): control group, model group, Xiaoyan Decoction group, rhG-CSF group, and Xiaoyan Decoction combined with rhG-CS group. Lewis-tumor mice in Xiaoyan Decoction group were ig given Xiaoyan Decoction 18.2 g/(kg·d) twice daily, for 7 d. Lewis-tumor mice in rhG-CS group were treated by sc injection of rhG-CSF. The routine blood test, immunological functions of peripheral blood and cytokines (IL-3, IL-6, EPO, and GM-CSF) in serum were detected. Results: The bone marrow suppression model was successfully established. Compared with control group, the levels of WBC, RBC, Hb, and PLT in peripheral blood of model group were decreased, especially in WBC (P 0.05). The concentration of IL-3 and IL-6 was dramatically reduced in model group compared with control group (P < 0.05). And it showed an obviously increasing trend in IL-3, IL-6, EPO, and GM-CSF levels in Xiaoyan Decoction group and combined group, as well as an increase in IL-3, IL-6 levels in rhG-CS group (P < 0.05). Conclusion: It shows that Xiaoyan Decoction combined with rhG-CS can improve the level of WBC and immunological functions of peripheral blood in bone marrow suppression model mice. And the effect of combination of both drugs is more significant. This might be related to the effect of promoting the production of serum hematopoietic regulator such as IL-3, IL-6, EPO, and GM-CSF, and relieving chemotherapy-induced bone marrow suppression.

Antitumor effect of malaria genetically attenuated sporozoites infection in a murine lewis lung cancer model / 重庆医学

Objective To learn whether plasmodium genetic attenuated sporozoites (GAS) can induce immunity against lung cancer ,in order to provide new ideas for the study of lung cancer vaccine .Methods Ther study was divided into two groups respec‐tively ,experimental group received intravenous injection of genetically attenuated sporozoites to immunize C57BL/6J mice and con‐trol group injection of phosphate buffer solution (PBS);after 14 days ,we subcutaneously inoculated lewis lung cancer (LLC) cells , calipers was used to measure tumor size .Immunohistochemical staining was detected tumor proliferation ,apoptosis ,and angiogene‐sis .Results There was statistically significant in tumor size .Immunohistochemical staining revealed that attenuated sporozoites in‐fection inhibited LLC eslls proliferation ,angiogenesis ,apoptosis .Conclusion The malaria attenuated sporozoites may provide a no‐vel strategy or therapeutic vaccine vector for anti‐lung cancer immune‐based therapy .

Antitumor effect of recombinant human endostatin combined with cisplatin on rats with transplanted Lewis lung cancer

Objective: To observe the antitumor effect and mechanism of recombinant human endostatin (Endostar) injection in tumor combined with intraperitoneal injection of cisplatin on subcutaneous transplanted Lewis lung cancer in rats. Methods: A total of 30 C57 rats were selected, and the monoplast suspension of Lewis lung cancer was injected into the left axilla to prepare the subcutaneous transplanted tumor models in the axilla of right upper limb. The models were randomly divided into Groups A, B, and C. Medication was conducted when the tumor grew to 400 mm3. Group A was the control group without any interventional treatment. Group B was injected with Endostar 5 mg kg-1 d-1 for 10 d. Group C was given the injection of Endostar 5 mg kg-1 d-1 combined with intraperitoneal injection of cisplatin 5 mg kg-1 d-1 for 10 d. All the rats in three groups were executed the day after the 10 d medication and the tumor was taken off for measurement of volume and mass changes and calculation of antitumor rate, after which the vascular endothelial growth factor (VEGF) concentration in rats' plasma was determined by ELISA. The tumor tissues were cut for the preparation of conventional biopsies. After hematoxylin-eosin staining, the pathologic histology was examined to observe the structures of tumor tissues, VEGF score and microvessel density (MVD) in each group. Results: The volume and mass of tumor in Groups B and C were significantly lower than Group A (P < 0.05) while the tumor volume and mass in Group C were significantly lower than Group B (P < 0.05). The antitumor rate in Group C was significantly higher than Group B (P < 0.05), but the tumor VEGF score, MVD and plasma VEGF level in Group C were significantly lower than Groups A and B (P < 0.05). In Group B, the tumor VEGF score, MVD and plasma VEGF level were significantly lower than Group A (P < 0.05). The microscopic image of Group C showed that its number of active tumor cells and the blood capillary around tumor was significantly smaller than that of Groups A and B, and meanwhile atrophy and liquefactive necrosis were seen in local tumor. Conclusions: Endostar injection combined with intraperitoneal injection of cisplatin is effective in reducing tumor VEGF score and MVD of transplanted tumor tissues in rats with Lewis lung cancer to obstruct the nutrient supply of tumor cells and kill tumor cells, so that the inhibition of tumor cell proliferation and metastasis can be achieved with a remarkable effect.

Effect of NF- κ B inhibitor PDTC on VEGF and endostatin expression of mice with Lewis lung cancer

Objective: To investigate the effects of NF- κ B inhibitor pyrrolidine dithiocarbamate hydrochloride (PDTC) on vascular endothelial growth factor (VEGF) and endostatin expression in mice with Lewis lung cance; and its mechanism. Methods: Mice survival rate and anti-tumor effects were observed in different concentrations of NF- κ B inhibitor PDTC after the Lewis lung cancer mice model was established. VEGF and endostatin expressions were detected by immunohistochemical assay. Results: Lewis lung cancer was be inhibited by 0.5 mg/kg, 1.5 mg/kg and 3.0 mg/kg of NF- κ B inhibitor PDTC (. P<0.05). Microvessel density (MVD) in 0.5 mg/kg, 1.5 mg/kg and 3.0 mg/kg NF- κ B inhibitor PDTC groups were significantly lower than the control group (. P<0.05). Immunohistochemical assay results showed that VEGF and endostatin expressions in the 0.5 mg/kg, 1.5 mg/kg and 3.0 mg/kg NF-. κ B inhibitor PDTC groups were significantly lower than the control group (. P<0.05). Western blot results also showed that NF- κ B inhibitor PDTC could inhibit VEGF and endostatin expressions in tumor tissues. Conclusions: NF- κ B inhibitor PDTC can inhibit tumor formation and reduce tumor angiogenesis in mice with Lewis lung cancer; and its mechanism maybe associated to VEGF and endostatin down-regulation.

Antitumor effect of recombinant human endostatin combined with cisplatin on rats with transplanted Lewis lung cancer

OBJECTIVE@#To observe the antitumor effect and mechanism of recombinant human endostatin (Endostar) injection in tumor combined with intraperitoneal injection of cisplatin on subcutaneous transplanted Lewis lung cancer in rats.@*METHODS@#A total of 30 C57 rats were selected, and the monoplast suspension of Lewis lung cancer was injected into the left axilla to prepare the subcutaneous transplanted tumor models in the axilla of right upper limb. The models were randomly divided into Groups A, B, and C. Medication was conducted when the tumor grew to 400 mm(3). Group A was the control group without any interventional treatment. Group B was injected with Endostar 5 mg kg(-1) d(-1) for 10 d. Group C was given the injection of Endostar 5 mg kg(-1) d(-1) combined with intraperitoneal injection of cisplatin 5 mg kg(-1) d(-1) for 10 d. All the rats in three groups were executed the day after the 10 d medication and the tumor was taken off for measurement of volume and mass changes and calculation of antitumor rate, after which the vascular endothelial growth factor (VEGF) concentration in rats' plasma was determined by ELISA. The tumor tissues were cut for the preparation of conventional biopsies. After hematoxylin-eosin staining, the pathologic histology was examined to observe the structures of tumor tissues, VEGF score and microvessel density (MVD) in each group.@*RESULTS@#The volume and mass of tumor in Groups B and C were significantly lower than Group A (P < 0.05) while the tumor volume and mass in Group C were significantly lower than Group B (P < 0.05). The antitumor rate in Group C was significantly higher than Group B (P < 0.05), but the tumor VEGF score, MVD and plasma VEGF level in Group C were significantly lower than Groups A and B (P < 0.05). In Group B, the tumor VEGF score, MVD and plasma VEGF level were significantly lower than Group A (P < 0.05). The microscopic image of Group C showed that its number of active tumor cells and the blood capillary around tumor was significantly smaller than that of Groups A and B, and meanwhile atrophy and liquefactive necrosis were seen in local tumor.@*CONCLUSIONS@#Endostar injection combined with intraperitoneal injection of cisplatin is effective in reducing tumor VEGF score and MVD of transplanted tumor tissues in rats with Lewis lung cancer to obstruct the nutrient supply of tumor cells and kill tumor cells, so that the inhibition of tumor cell proliferation and metastasis can be achieved with a remarkable effect.

Effect of NF- κ B inhibitor PDTC on VEGF and endostatin expression of mice with Lewis lung cancer

OBJECTIVE@#To investigate the effects of NF- κ B inhibitor pyrrolidine dithiocarbamate hydrochloride (PDTC) on vascular endothelial growth factor (VEGF) and endostatin expression in mice with Lewis lung cance; and its mechanism.@*METHODS@#Mice survival rate and anti-tumor effects were observed in different concentrations of NF- κ B inhibitor PDTC after the Lewis lung cancer mice model was established. VEGF and endostatin expressions were detected by immunohistochemical assay.@*RESULTS@#Lewis lung cancer was be inhibited by 0.5 mg/kg, 1.5 mg/kg and 3.0 mg/kg of NF- κ B inhibitor PDTC (P<0.05). Microvessel density (MVD) in 0.5 mg/kg, 1.5 mg/kg and 3.0 mg/kg NF- κ B inhibitor PDTC groups were significantly lower than the control group (P<0.05). Immunohistochemical assay results showed that VEGF and endostatin expressions in the 0.5 mg/kg, 1.5 mg/kg and 3.0 mg/kg NF-κ B inhibitor PDTC groups were significantly lower than the control group (P<0.05). Western blot results also showed that NF- κ B inhibitor PDTC could inhibit VEGF and endostatin expressions in tumor tissues.@*CONCLUSIONS@#NF- κ B inhibitor PDTC can inhibit tumor formation and reduce tumor angiogenesis in mice with Lewis lung cancer; and its mechanism maybe associated to VEGF and endostatin down-regulation.

Antitumor effect of recombinant human endostatin combined with cisplatin on rats with transplanted Lewis lung cancer

Objective:To observe the antitumor effect and mechanism of recombinant human endostatin (Endostar) injection in tumor combined with intraperitoneal injection of cisplatin on subcutaneous transplanted Lewis lung cancer in rats.Methods:A total of 30 C57 rats were selected, and the monoplast suspension of Lewis lung cancer was injected into the left axilla to prepare the subcutaneous transplanted tumor models in the axilla of right upper limb. The models were randomly divided into Groups A, B, and C. Medication was conducted when the tumor grew to 400 mm3. Group A was the control group without any interventional treatment. Group B was injected with Endostar 5 mg.kg-1.d for 10 d. Group C was given the injection of Endostar 5 mg.kg-1.d combined with intraperitoneal injection of cisplatin 5 mg.kg-1.d for 10 d. All the rats in three groups were executed the day after the 10-d medication and the tumor was taken off for measurement of volume and mass changes and calculation of antitumor rate, after which the vascular endothelial growth factor (VEGF) concentration in rats’plasma was determined by ELISA. The tumor tissues were cut for the preparation of conventional biopsies. After hematoxylin-eosin staining, the pathologic histology was examined to observe the structures of tumor tissues, VEGF score and microvessel density (MVD) in each group. Results:The volume and mass of tumor in Groups B and C were significantly lower than Group A (P< 0.05) while the tumor volume and mass in Group C were significantly lower than Group B (P < 0.05). The antitumor rate in Group C was significantly higher than Group B (P < 0.05), but the tumor VEGF score, MVD and plasma VEGF level in Group C were significantly lower than Groups A and B (P < 0.05). In Group B, the tumor VEGF score, MVD and plasma VEGF level were significantly lower than Group A (P < 0.05). The microscopic image of Group C showed that its number of active tumor cells and the blood capillary around tumor was significantly smaller than that of Groups A and B, and meanwhile atrophy and liquefactive necrosis were seen in local tumor.Conclusions:Endostar injection combined with intraperitoneal injection of cisplatin is effective in reducing tumor VEGF score and MVD of transplanted tumor tissues in rats with Lewis lung cancer to obstruct the nutrient supply of tumor cells and kill tumor cells, so that the inhibition of tumor cell proliferation and metastasis can be achieved with a remarkable effect.