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Objective To investigate the effect and mechanism of miR-20a-5p on human nephroblastoma cell line WiT49 transplanted tumor in nude mice.Methods The gene expression chip was downloaded from GEO database,and the differential gene miR-20a-5p was obtained by GEO2R.The NF-κB gene was positively correlated with the expression of miR-20a-5p through cBioPortal database.The target gene of miR-20a-5p was predicted to be NFKBIB of the NF-κB transcription factor suppressor protein family by targetscan database,and was verified by dual luciferase assay.Nephroblastoma cell line WiT49 was cultured in vitro and transfected into WiT49 cells with lentiviral vectors constructed with miR-20a-5p mimics and its suppressor gene.Twelve nude mice were randomly divided into three groups:WiT49 model group,WIT49-miR-20a-5p overexpression group and WIT49-miR-20a-5p knockdown group.The tumor mass and volume of each group were detected by tumor formation experiment in nude mice.real time fluorescent quantitative polymerase chain reaction(qRT-PCR)was used to detect the expression of miR-20a-5p,NFKBIB and NF-κB in each group;CCK-8 cell proliferation assay was used to verify the proliferation of tumor cells in each group.Results miR-20a-5p is highly expressed in nephroblastoma and is positively correlated with the expression of NF-κB.miR-20a-5p and NFKBIB have mutual binding sites and binding effects.In the tumor formation experiment of nude mice,the tumor volume and mass of WIT49-miR-20a-5P overexpression group were significantly increased compared with WiT49 model group,and the difference was statistically significant(P<0.05).In the qRT-PCR test,the expressions of miR-20a-5p and NF-κB in the WIT49-miR-20a-5p overexpression group were higher than those in the WiT49 model group,and NFKBIB expression in the WIT49-miR-20a-5p overexpression group was lower than that in the WiT49 model group,with statistical significance(P<0.05).CCK-8 cell proliferation assay showed that the absorbance of WIT49-miR-20a-5p overexpression group at 24 and 48 hours was higher than that of WiT49 model group,and the absorbance of WIT49-miR-20a-5p knockdown group at 24,48 and 72 hours was lower than that of WiT49 model group,and the difference was statistically significant(P<0.05).Conclusion miR-20a-5p may promote the growth of human nephroblastoma cell WiT49 transplanted tumor in nude mice by regulating NFKBIB activation of NF-κB pathway.
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ObjectiveTo observe the primary tumor of tree shrews and to provide a basis for studying the pathogenesis and prevention of trichoepithelioma. MethodsA tumor was discovered in the chest and abdomen of a tree shrew during natural cultivation. The tree shrew was anesthetized, and the tumor was surgically removed. Hematoxylin and eosin (HE) staining and immunohistochemical staining were performed on the tumor tissue after paraffin section, and the tumor cells were isolated and cultured by passage. The isolated tumor cells were subcutaneously injected into healthy tree shrews and nude mice. The tumorigenesis of tumor cells in vivo was observed once a day, with nude mice continuously observed for 2 months and tree shrews observed for more than 6 months. ResultsHE staining showed that the basal cells in the dermis were arranged as a whole, like a string of petals, forming nests and stripe-like structures with clear boundaries. The observation results after magnification revealed that the tumor cells were arranged in a pallisade-like and basal pattern, with deep nuclear staining and minimal cytoplasmic. Immunohistochemical staining showed the high expression of CK protein and low proportion expression of ki-67 protein in tumor cells, as well as the high expression of vimentin and low expressions of Bcl2 and CD10 in tumor cell mesenchyme. The isolated tumor cells grew well in DMEM medium containing 10% fetal bovine serum and could be cultured by passage, but no tumor formation was observed in healthy tree shrews and nude mice inoculated with tumor cells. ConclusionCombined with the location of the tumor, overall morphology, HE staining, and immunohistochemical results, the thoracoabdominal mass of the tree shrew was diagnosed as a trichoepithelioma.
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C17 is an orally available anti-tumor compound inhibiting cancer stem cell (CSC). In this study, a stable, sensitive and simple liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was established and validated, and was further applied to a pharmacokinetic study in nude mice receiving C17 by gavage. Using propranolol as the internal standard, the plasma samples were pre-treated by precipitation with methanol and analyzed on an Intersil C8-3 column (100 mm × 2.1 mm, 3 μm), and gradient elution was performed with a mobile phase consisting of 0.1% formic acid aqueous and solution mixed up by 90% isopropanol and 10% acetonitrile. The analyte was detected by a triple quadrupole tandem mass spectrometer, and multiple reaction monitoring was employed to select C17 at m/z 439.3/247.1 and propranolol at m/z 260.2/116.2 in the positive ion mode. The calibration curves were linear (r > 0.995) over the range of 5-800 ng·mL-1. The intra- and inter-day precisions and accuracies were 7.42%-13.22% and -8.99%-8.81% respectively. The method was successfully applied to a PK study in nude mice administered with a single oral dose of 50 mg·kg-1 C17, and the PK data were analyzed with non-linear mixed effect model (NONMEM). Two separated absorption peaks were found in the PK curve of C17, and a two-compartment model with two sequential first-order absorption rate was utilized to describe the PK properties of C17, and the model could provide insights into the physiological process and exposure of C17 in nude mice. All animal experiments were in strict accordance with the regulations of the Biomedical Ethics Committee of Peking University.
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Objective:To explore the mechanism of fat formation induced by PLA-adipose derived stem cells (ADSC) and LAF-ADSC cells, and to provide a new direction and idea for cell-assisted granular fat transplantation.Methods:The liposuction operation of normal human body was performed in the plastic surgery operating room of Chengdu Badachu Medical Cosmetic Hospital. The liposuction obtained through the operation was placed still, and the upper granular adipose tissue was digested, isolated, cultured and identified by PLA-ADSC; the obtained lower liquid tissue was centrifuged, and the precipitate was digested, isolated, cultured and identified by LAF-ADSC; the differentiation characteristics and growth ability of PLA-ADSC and LAF-ADSC were compared. Then animal experiments were carried out: PLA-ADSC and LAF-ADSC were mixed with matrix glue Matrigel and transplanted into nude mice as experimental group 1 and experimental group 2. Matrix glue Matrigel was transplanted into nude mice as control group. The lipogenic ability of the three groups in nude mice was compared.Results:The cell experiment showed that the cells extracted from the static granular fat and the lower filtrate sediment could adhere to the wall and grew and subcultured smoothly. The cells from the above two different sources expressed CD44, CD73 and CD105, with positive rates of 99.5%, 99.99% and 99.7% respectively, while CD19, CD31 and CD45 were negative. The results of lipogenic induction and differentiation showed that there were lipid droplets in the cytoplasm, and the lipid droplets were orange red after cell oil red O staining. The results of animal experiment showed that three months after transplantation, the average graft volume of experimental group 1 was (0.070±0.009) cm 3, that of experimental group 2 was (0.067±0.007) cm 3, and that of the control group was (0.009±0.005) cm 3. The difference between the graft volume of experimental group 1 and experimental group 2 and the control group was statistically significant ( t=26.522 and t=37.183, all P<0.01), There was no significant difference in graft volume ( t=1.250, P>0.05). The wet weight of grafts in experimental group 1 was (0.200±0.021) g, that in experimental group 2 was (0.175±0.019) g, and that in the control group was (0.129±0.012) g, there was significant difference between experimental groups 1 and 2 and the control group ( t=11.601 and t=9.978, P<0.05), but there was no significant difference between experimental group 1 and experimental group 2 ( t=3.650, P>0.05). After oil red O staining, the grafts in experimental groups 1 and 2 were generally orange yellow, and the control group was scattered light yellow. The expression of CD31 in the experimental groups 1 and 2 was positive, and the expression of CD31 in control group was negative. Conclusions:Active ADSCs can be extracted from the granular fat layer and the lower filtrate of the static fat aspirate, and the ADSCs from both sources have good lipogenic ability in nude mice.
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【Objective】 To investigate the effect of polymerized human cord hemoglobin (PolyCHb) on the chemosensitivity of human breast cancer MCF-7 cell subcutaneous xenografts in nude mice and its mechanism. 【Methods】 The MCF-7 cells in exponential growth phase were collected and made into suspension cells at a density of 5×107 cells/mL.Subsequently, the cells were inoculated subcutaneously in the right limb of 18 BALB/c-nu nude mice with 0.2 mL cells per mouse to establish subcutaneous xenograft.When the tumor volume reached about 100 mm3, they were randomly divided into chemotherapy group: doxorubicin 5 mg·kg-1, once/week; chemotherapy + PolyCHb group: in addition to doxorubicin (chemotherapy group), PolyCHb 600 mg·kg-1, 3 times/week; the control group: normal saline 90 mg·kg-1, once/week; all were injected through tail vein continuously for 4 weeks.From the day of injection (d 0), the tumor volume of each group of nude mice was measured every 3 days, and the tumor growth curves were drawn accordingly.After 38 days, the tumor growth observation was completed.The tumor was removed and weighed to calculate the tumor inhibition rate.HE staining, immunohistochemistry and TUNEL method were used to observe the pathological changes of tumor tissue, detect the expression of HIF-1α, and detect tumor cell apoptosis respectively.The content of reactive oxygen species (ROS) of each group was determined by fluorescence staining. 【Results】 The tumor volume (mm3) of chemotherapy + PolyCHb group, chemotherapy group and the control group at day 38 were 196.35±103.45 vs 316.29±62.88 vs 519.42±177.33 (P<0.05), and the tumor inhibition rate (%) of chemotherapy + PolyCHb treatment group and chemotherapy group was 62.20 vs 39.11, respectively.HE staining and TUNEL detection showed that cell necrosis and apoptosis in the growth area of tumor tissue increased in chemotherapy + PolyCHb group.Immunohistochemistry and fluorescence staining showed that HIF-1α expression in chemotherapy + PolyCHb group decreased and reactive oxygen species (ROS) content increased. 【Conclusion】 PolyCHb increases the chemosensitivity of subcutaneous xenograft in nude mice with breast cancer, and its mechanism may be related to the increase of ROS in tumor tissue and the promotion of tumor cell apoptosis.
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Recent studies have shown that miR-338-3p plays an important role in the proliferation and invasion of lung cancer, but whether miR-338-3p regulates lung cancer proliferation and invasion through targeting ring finger protein 121 (RNF121) is still unclear. In order to explore its mechanism, the normal lung cell line MRC-5 and the non-small cell lung cancer line A549 were cultured in vitro. Using qRTPCR and Western blotting detection, we found that the expression of miR-338-3p in A549 lung cancer cells was lower than that in MRC-5 cells, while RNF121 expression increased (P0. 05). In summary, miR-338-3p can target the expression of RNF121 to inhibit the proliferation and invasion of A549 cells and inhibit the growth of subcutaneous transplanted tumors in nude mice. RNF121 is expected to become a new target for the treatment of non-small cell lung cancer.
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Aim To explore the effects of EGCG on xenografts of ovarian cancer in nude mice and its possible mechanism. Methods Nude mice xenografts of ovarian cancer SK0V3 cells were established and divided into five groups after tumor formation, in which three groups were given EGCG (10, 30, 50 mg • k g-
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OBJECTIVE@#To establish an animal model with malignant tumor in the skull base-infratemporal region, and to explore the role of iodine staining technique in identifying tumor tissues with Micro-CT data.@*METHODS@#Sedation anesthesia was carried out on 12 BABL/c nude mice using inhaled isoflurane, and then WSU-HN6 cells that cultured and immortalized from human tongue squamous cell carcinoma were injected into the right infratemporal fossa via the submandibular area. The procedure was carried out under ultrasonographic guidance. The nude mice were sacrificed after 3 weeks observation. The head specimens were fixed and scanned by Micro-CT, and repeated scans were performed after staining with 3.75% compound iodine solution. Following decalcification in 20% EDTA for 2-4 weeks, the head specimens were embedded and sectioned. Hematoxylin and eosin staining and Pan-Keratin immunohistochemical staining were carried out. Bright-field microscopy and stereomicroscopy were used to visualize. The Micro-CT data were analyzed using iPlan software (Brainlab).@*RESULTS@#Non-traumatic ultrasonography was used to guide HN-6 cells injection and confirm skull-base tumor formation in all the animals. Ultrasonographic guidance reduced the risk of cervical vessel injury when transferring tumor cells into the skull base space. An obvious asymmetrical appearance was detected via ultrasonography 3 weeks after tumor cell injection. The Micro-CT analysis showed that the bone was obviously damaged on the right side of the skull base, but the soft tissue image was unrecognizable. After four days staining with compound iodine solution, the morphology of the tumor and surrounding soft tissue could be clearly identified. Hematoxylin and eosin staining showed the tumor formation of the right infratemporal fossa region accompanied by bone destruction. Human keratin immunohistochemical staining showed that the tumor tissue originated from human squamous cell carcinoma, and the polynuclear osteoclasts could be seen at the margin of the skull base bone resorption.@*CONCLUSION@#The animal model with malignant tumor in the skull base-infratemporal region could be successfully established via submandibular injection under ultrasound-guidance. Bone changes of the skull were easily observed on Micro-CT, but the tumor counter was not able to be distinguished from surrounding soft tissue. The 3.75% compound iodine staining of the head specimen could help discern the tumor and surrounding soft tissue in more details.
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Animais , Camundongos , Carcinoma de Células Escamosas/diagnóstico por imagem , Fossa Infratemporal , Iodo , Camundongos Nus , Base do Crânio , Coloração e Rotulagem , Neoplasias da Língua , Microtomografia por Raio-XRESUMO
【Objective】 To investigate the effect of Polymerized human cord hemoglobin (PolyCHb) on the sensitivity of hepatocellular carcinoma grafts to lumvalatinib in nude mice. 【Methods】 Hep3B hepatoma cells were subcutaneously transplanted in 18 nude mice to establish tumor graft model. Mice were randomly divided into 3 groups: control group (the saline 90 mg·kg-1·d-1), monotherapy group (Lenvatinib10 mg·kg-1·d-1), and sensitized group (Lenvatinib mg·kg-1·d-1, polyCHB 600 mg/kg twice a week) for 28 days. The tumor volume was measured regularly and the growth curve was drawn. On day 29, the nude mice were sacrificed, the tumor was stripped and weighed, and the pathomorphological differences of each group were evaluated by HE section staining. The expression levels of hypoxia-inducing factor (HIF-1α), CD34, VEGF, CD44, MMP-9, and Glut-1 in tumor tissues of each group were determined by immunohistochemistry. The content of reactive oxygen species (ROS) in tumor tissues of each group was determined by dihydroethyl ingot method. 【Results】 The tumor growth rate and tumor volume in the sensitized group decreased significantly compared with the control group and the solo drug group. On day 29, the tumor volumes of the control group, the monotherapy group and the sensitization group were (2 076.46±350.25)mm3, (1 035.96±84.16)mm3 and (892.66±104.46)mm3, respectively. Tumor weight was (1.61±0.52)g, (0.45±0.10)g, and (0.34±0.13)g, respectively. Immunohistochemical score of HIF-1α was 75±23 vs 45±18 vs 18±11, VEGF was 52±8 vs 67±16 vs 35±4, CD34 was 40±7 vs 50±13 vs 28±7, CD44 was 37±15 vs 30±7 vs 15±3, Glut-1 was 74±41 vs 51±30 vs 14±18, MMP-9 was 51±7 vs 62±20 vs 33±3, respectively(P<0.05). The malignant degree of the sensitized group was decreased by HE section staining, which was significantly lower than that of the solo drug group and the control. The ROS content in the sensitized group was higher than that in the solo drug group and the control. 【Conclusion】 PolyCHb can reduce the expression of HIF-1α and its downstream pathway related molecules by increasing oxygenation of hepatocellular carcinoma tissues in nude mice, delay tumor growth and reduce tumor volume in a certain period, thus increase the therapeutic effect of lenvalatinib on hepatocellular carcinoma grafts in tumor bearing nude mice models.
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@#[Abstract] Objective: To explore the application value of human IL-15 transgenic NCG mice (NCG-hIL-15 mice) in preclinical evaluation of chimeric antigen receptor modified NK (CAR-NK) cell therapy for tumor treatment. Methods: qPCR and WB were performed to detect the expression of human IL-15 in the bone marrow and main organs (spleen, liver, lung, kidney and pancreas) of transgenic mice. After being transfused with human PBMC-derived NK (PB-NK) cells, the NCG-hIL-15 mice and control NCG mice were continuously monitored for the in vivo amplification of NK cells and the changes in body weight and survival time. Flow cytometry was used to detect the differential expressions of activated receptors and inhibitory receptors in amplified NK cells. WB was used to detect the expressions of perforin and granzyme-B. NCG-hIL-15 mice or NCG mice bearing MIAPaca-2 cell transplanted tumor were treated with anti-MUC1-CAR-NK cell reinfusion; then, the CAR-NK cell survival in different groups of mice was detected by Flow cytometry, and the survival time of tumor bearing mice was recorded and tumor growth was detected by in vivo imaging. Results: The results indicated that PB-NK cells could proliferate stably within 10 weeks in NCG-hIL-15 mice without obvious graft versus host diseases (GVHD) during the observation period. The in vivo-expanded human NK cells maintained the original expression patterns of various surface molecules, including KARs and KIRs. Compared with the NK cells in NCG mice, the NK cells in NCG-hIL-15 mice contained significantly higher amounts of granzyme-B and perforin (all P<0.05). CAR-NK cells showed significantly increased survival rate and stronger tumor-inhibitory effect in NCG-hIL-15 mice as compared with those in control NCG mice, resulting in significantly prolonged survival in NCG-hIL-15 mice (all P<0.01). Conclusion: NCG-hIL-15 mouse model has potential application value in preclinical trial and biological evaluation of NK cell-based immunotherapy.
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Sempervirine, a yohimbane-type alkaloid isolated from Gelsemium elegans, was found to significantly inhibit the cellular proliferation of U251 cells in vitro and in vivo in a dose-dependent manner. U251 cells were treated with 0-16 μmol·L-1 of sempervirine for 24, 48 or 72 h. An MTT assay and clone formation assay were used to investigate cell survival and clone formation. Hoechst staining and Annexin V-FITC/PI staining were used to measure cell apoptosis. The expression of PI3K, AKT, p-AKT, Bax, Bcl-2, caspase-3 and cleaved caspase-3 was determined by Western blot analysis. The antitumor effect of sempervirine in vivo was investigated by inoculating nude mice with U251 cells. All animal experiments were in strict accordance with the regulations of the Biomedical Ethics Committee of Fujian Medical University (Fujian, China). The results show that sempervirine significantly inhibits the proliferation and induces the apoptosis of U251 cells, promotes cleavage of caspase-3, down-regulates the protein expression of PI3K and Bcl-2/Bax, and inhibits phosphorylation of AKT in vitro. Intraperitoneal injection of 4 or 8 mg·kg-1·day-1 of sempervirine inhibits U251 cells tumor growth in the xenograft nude mice, and tumor weight decreased by 44.76% and 61.26%, respectively. Our study shows that sempervirine significantly inhibits the proliferation of U251 cells in vitro and in vivo, laying a foundation for further research and development of its anti-glioma effect.
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RU486 (mifepristone), a glucocorticoid and progesterone receptor antagonist, has been reported to exert antiproliferative effects on tumor cells. Experiments were performed to analyze the effects of RU486 on the proliferation of the human neuroblastoma, both in vitro and in vivo, using the human neuroblastoma SK-N-SH cell line. The exposure in vitro of SK-N-SH cells to RU486 revealed a dose-dependent inhibition of 3H-thymidine incorporation due to a rapid but persistent inhibition of MAPKinase activity and ERK phosphorylation. A significant decrease of SK-N-SH cell number was evident after 3, 6, and 9 days of treatment (up to 40% inhibition), without evident cell death. The inhibitory effect exerted by RU486 was not reversed by the treatment of the cells with dexamethasone or progesterone. Moreover, RU486 induced a shift in SK-N-SH cell phenotypes, with an almost complete disappearance of the neuronal-like and a prevalence of the epithelial-like cell subtypes. Finally, the treatment with RU486 of nude mice carrying a SK-N-SH cell xenograft induced a strong inhibition (up to 80%) of tumor growth. These results indicated a clear effect of RU486 on the growth of SK-N-SH neuroblastoma cells that does not seem to be mediated through the classical steroid receptors. RU486 acted mainly on the more aggressive component of the SK-N-SH cell line and its effect in vivo was achieved at a concentration already used to inhibit oocyte implantation.
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Humanos , Animais , Coelhos , Neuroblastoma/tratamento farmacológico , Progesterona , Mifepristona/farmacologia , Glucocorticoides , Camundongos NusRESUMO
This study was designed to investigate the effect of dihydromyricetin (DHM) on inducing apoptosis of ovarian cancer cells A2780 through endoplasmic reticulum stress (ERS) pathway and the mechanisms involved in vitro and in vivo. A2780 cells were treated with different concentrations of DHM, and the protein expression levels of glucose-regulated protein 78 (GRP78) which is related to ERS increased, apoptotic proteins C/EBP-homologous protein (CHOP), and cysteinyl aspartate specific proteinase-12 (caspase-12) elevated. After pretreatment with ERS inhibitor, 4-phenyl butyric acid (4-PBA), following the intervention with DHM, the A2780 cell viability decreased and apoptotic rate increased. All animal welfare and experimental procedures were approved by the Animal Ethics Committee of Chongqing Medical University. Intraperitoneal injection of DHM suspension into nude mice with ovarian cancer could significantly inhibit the growth of transplanted tumor in vivo, increase the protein expression levels of GRP78, CHOP, and caspase-3. Moreover, swollen and broken endoplasmic reticulum could be observed in tumor tissues, suggesting that DHM intervention induces apoptosis mediated by ERS. The results indicated that DHM could induce apoptosis of ovarian cancer cells and inhibit the growth of transplanted tumors in nude mice, which might be related to the activation of ERS pathway.
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Objective: To explore the effects of up- and down-regulation of circadian clock gene Bmal1 on the growth and radiation sensitivity of nasopharyngeal carcinoma after CNE1 xenograft in nude mice. Methods: We produced four groups of cells using lentiviral transfection: cells overexpressing CNE1 (CNE1OE), negative control in which there was no overexpression of CNE1 (OENC), cells with short hairpin RNAi (CNE1sh3), and RNAi negative cells (CNE1shNC). We investigated the expression of Bmal1 protein in the aforementioned groups with Western blot. After subcutaneously injecting the four groups of cells in nude mice, the size of the xenograft was measured. Subsequently, the xenografts were irradiated with 15 Gy at 6 MeV, and variation in the xenograft volume was recorded. mRNA and protein expression levels of Bmal1, p53, and p21 in the xenograft were measured with RT-PCR and Western blot, respectively. Results: The CNE1OE group highly expressed Bmal1 protein whereas the CNE1sh3 group was silenced by RNAi as shown with the Western blot, indicating successful transfection. The xenograft in the nude mice developed well. The CNE1OE xenograft volume was lower than that of the CNE1OENC xenograft, whereas the CNE1sh3 xenograft was larger than the CNE1shNC xenograft (P<0.05). CNE1OE, CNE1OENC, and CNE1shNC xenograft volumes shrank after being irradiated (t=4.32, 5.38, 5.16, respectively; P<0.05) and the effect was the highest in the CNE1OE group. However, there was almost no variation in xenograft volume in the CNE1sh3 group. The relative amounts of mRNA and protein of Bmal1, P53, and P21 were higher in the CNE1OE group than in the CNE1OENC group,while they were lower in the CNE1sh3 group compared to the CNE1shNC group (P<0.05). Conclusions: Overexpression of Bmal1 inhibited the growth of the CNE1 xenograft in nude mice and enhanced its radiation sensitivity whereas silencing the Bmal1 gene by RNAi promoted the growth of the xenograft and led to radiation resistance. We believe that Bmal1 overexpression leads to P53 and P21 overexpression, thereby inhibiting the growth of the xenograft.
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@#[Abstract] Objective: To construct and verify the anti-tumor activity of chimeric antigen receptor (CAR) modified NK-92 cells (CAR-NK-92 cells) targeting prostate stem cell antigen (PSCA) in cervical cancer. Methods: Lentiviral vector expressing CAR targeting PSCA was constructed, and PSCA CAR-NK-92 cells were obtained by lentivirus transfection. The expression of PSCA in human cervical cancer cells was determined by Flow cytometry and Western blotting. The killing effect of PSCA CAR-NK-92 cells against cervical cancer cells was verified by co-incubation of effector and target cells in vitro, and the tumor inhibitory ability of PSCA CAR-NK-92 cells was verified with the nude mice xenograft model in vivo. Results: PSCA CAR-NK-92 cells were successfully constructed. PSCA was highly expressed in human cervical cancer Hela and MS751 cells (all P<0.01). In vitro co-incubation results showed that PSCA CAR-NK-92 cells could lyse PSCA+ cervical cancer transplanted tumor in a dose-dependent manner. In vivo anti-tumor data showed that PSCA CAR-NK-92 cells significantly inhibited the growth of cervical cancer cells compared with NK-92 cells transfected with vehicle vectors (P<0.01). In addition, PSCA CAR-NK-92 cells could effectively infiltrate tumor tissues and promote the secretion of anti-tumor cytokines TNF-α and IFN-γ (all P<0.01). Conclusion: The CAR-NK-92 targeting PSCA shows good anti-tumor effect on PSCA+ tumor cells both in vitro and in vivo, and has potential to be a therapeutic strategy for cervical cancer.
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Objective:To study the antitumor effect of Xihuangwan on A549 lung cancer nude mice in inflammatory microenvironment, and explore the effect of Xihuangwan on the expressions of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) inflammatory bodies and their products in serum and tumor tissue of A549 lung cancer nude mice. Method:The lung cancer A549 cell model was established in nude mice with lung cancer, and the lung cancer A549 cell model was established in inflammatory microenvironment by adding lipopolysaccharide (LPS) + adenosine triphosphate (ATP) to the culture medium. After modeling, the rats were randomly divided into blank group (equal volume of normal saline), positive drug control group (MCC950 solution, 0.79 g·kg-1), and low, medium, high-dose Xihuangwan groups (0.39, 0.78, 1.95 g·kg-1). The rats were administered orally once a day for 21 days, and then sacrificed. The tumor tissues were stripped to measure the tumor body. The expressions of NLRP3, malondialdehyde(MDA), interleukin (IL)-1β, IL-18 and NLRP3 were detected by Western blot, and the levels of IL-1β, IL-18 and MDA were detected by enzyme linked immunosorbent assay(ELISA). Result:Compared with the blank group, the tumor inhibition rates in the positive drug control group and the low, medium and high dose Xihuangwan group were 39.21%, 31.72%, 42.24% and 55.68%. ELISA showed that the high-dose Xihuangwan group could significantly reduce the expressions of MDA, IL-1β and IL-18 in the serum of nude mice (P<0.05). Western blot showed that the high-dose Xihuangwan group could effectively reduce the protein expressions of MDA, IL-1β, IL-18 and NLRP3 in tumor of nude mice (P<0.05), the results of immunohistochemistry showed that the expression rate of NLRP3 in the tumor tissues of nude mice was reduced in the positive drug group and each dose of Xihuangwan group (P<0.05). Conclusion:Xihuangwan can inhibit the growth of tumor tissue of A549 cells bearing lung cancer in nude mice. The mechanism may be that it can inhibit the growth of tumor cells by inhibiting the expression of NLRP3 inflammatory bodies, IL-1β, IL-18, MDA tables, and then inhibiting the inflammatory microenvironment of tumor cells.
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Objective:To observe the effect of Zengmian Yiliu (ZMYL) formula and its effective components (PAWU) on the growth inhibition of ovarian cancer stem cell transplanted tumor in nude mice and the Notch signal receptor (Notch) / Notch signal ligand 1 (Jagged1) signal pathway in tumor tissue. Method:Ovarian cancer stem cells were cultured in serum-free suspension to establish the transplanted tumor model of ovarian cancer stem cells in nude mice, and then divided into model group, ZMYL group (36 g·kg-1), PAWU group (5.8 g·kg-1), cisplatin (DDP) group (2.5 g·kg-1), and PAWU (5.8 g·kg-1) + DDP group (2.5 mg·kg-1).After successful modeling, the drugs were given by gavage for 21 days.To observe the effect of Zengmian Yiliu decoction and its effective components on tumor weight in nude mice, the morphological changes of tumor cells were observed under light microscope, immunohistochemistry and real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)were used to detect the expressions of Notch 1, Jagged1, Hairy and enhancer of split 1 (Hes1) protein and mRNA in tumor tissues. Result:The tumor inhibition rates of ZMYL, PAWU, DDP and combination groups were 35.91%, 32.94%, 57.65% and 69.05%, respectively.Compared with the model group, the tumor weight of ZMYL group, PAWU group, DDP group and combination groups decreased significantly (P<0.05).Compared with PAWU group, the tumor weight of combination groups decreased significantly (P<0.05).Immunohistochemistry showed that compared with the model group, the positive expressions of Notch1, Jagged1 in ZMYL group, PAWU group, DDP group and combination groups were down-regulated (P<0.05),and the positive expressions of Hes1 in ZMYL group, DDP group and combination groups were down-regulated (P<0.05).Compared with combination groups, the positive expressions of Notch1, Jagged1 and Hes1 in ZMYL group, PAWU group, DDP group were up-regulated (P<0.05). Real time PCR showed that compared with the model group, the expressions of Notch1 mRNA in ZMYL group, PAWU group, DDP group and combination groups decreased significantly (P<0.05).Compared with model group, the expressions of Jagged1 and Hes1 mRNA in ZMYL group, PAWU group and combination groups decreased significantly (P<0.05).Compared with DDP group, the expressions of Notch1, Jagged1 and Hes1 mRNA in combination groups decreased significantly (P<0.05). Conclusion:The growth of ovarian cancer stem cells transplanted in nude mice can be inhibited by Zengmian Yiliu formula and its effective components.The effective components have a significant synergistic effect in the combination with cisplatin.Its mechanism is correlated to the inhibition of Notch/Jagged1 signaling pathway activation.
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Objective Establishment of orthotopic transplantation tumor model of human choriocarcinoma in nude mice. Methods Human choriocarcinoma cell line JAR was cultured and the single cell suspension was subcutaneously injected into five 8-week-old BALB / c nude mice to establish subcutaneous xenograft model. After subcutaneous tumor was formed in nude mice, the tumor tissue was taken under aseptic conditions and cut into 1 mm
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Objective To investigate the effect of interleukin( IL)-6 secreted by bone marrow stromal cells HS-5 on activity and apoptosis of human acute myeloid leukemia( AML) cells HL-60 and its possible mechanism. Methods HL-60 cells and HS-5 cells were cultured in vitro, and the co-culture system was established. Scanning electron microscope, ELISA, cell counting kit-8( CCK-8) , AnnexinV-FITC/PI, double-staining flow cytometry, Real-time PCR and Western blotting techniques were used to detect the changes of viability and apoptosis of HL-60 cells, respectively. HL-60 cells from different groups were inoculated into BALB/c nude mice to observe and record tumorigenesis. Results IL-6 secreted by bone marrow stromal cells HS-5 could enhance the viability and inhibit the apoptosis of HL-60 cells, and down-regulate the expression of pro-apoptotic gene Bax and and up-regulate the expression of anti-apoptotic gene Bcl-2 in HL-60 cells. HL-60 cells in co-culture group had the strongest tumorigenicity in BALB/c nude mice, while HL-60 cells alone had the weakest tumorigenicity. Conclusion Part of the mechanism by which bone marrow stromal cells HS-5 promote the proliferation of HL-60 cells and inhibit their apoptosis may be through the secretion of IL-6.
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Aim To investigate the effect of deguelin on the proliferation of non-small cell lung cancer cell line A549 and nude mice. Methods CCK-8 assay was used to detect the inhibition of deguelin on proliferation of SH-SY5Y cells; Hoechst stains and Annex-inV-FITC/PI double stained method were employed to observe the apoptotic morphology and apoptotic rate; flow cytometry was applied to determine the effect of deguelin on cell cycle of A549; tumor xenograft experiment and HE staining were conducted to investigate the effect of deguelin on growth of transplanted tumor in nude mice. Results Deguelin inhibited cell proliferation of A549 dose- A nd time-dependently; Hoechst stains and AnnexinV-FITC/PI double stained further confirmed that deguelin could induce the apoptosis of A549 cells, while deguelin blocked A549 cell cycle in G2/M phase in concentration-dependent manner. The nude mice xenograft model and HE staining experiments showed that deguelin significantly inhibited the growth of xenografts in A549 nude npce (P < 0. 01). Conclusions Deguelin has a significant inhibitory effect on non-small cell lung cancer cell line A549, pointing to a basis for the study of the antitumor activity of deguelin.