miR-937 regulates the proliferation and apoptosis via targeting APAF1 in breast cancer.

Background: Previous research had shown that an imbalance in cell proliferation and apoptosis is a vital mechanism for tumorigenesis and cancer progression that may directly influence biological behaviors of cancer. microRNAs are associated with the occurrence and development of tumors. This study aimed to explore the influence of miR-937 on breast cancer regulation of APAF1 expression. Methods: Cancer Genome Altas microarray analysis (fold change > 2, p<0.05) was used to verify differentially expressed microRNAs and RT-qPCR was used to detect miR-937 mRNA level in breast cancer. Cell viability and proliferation were measured using CCK8 and colony formation assays, respectively, after the miR-937 mimics/inhibitors and their negative control were transfected into MCF7 cells. The variations in cell cycle and apoptosis were examined using flow cytometry. DAVID database was used to perform GO enrichment analysis. We use dual luciferase report system to detect the effect of miR-937 on the transcriptional activity of APAF1. APAF1 protein level was determined by Western blot assay. Results: miR-937 was up-regulated in breast cancer cell lines and high miR-937 expression is associated with a poorer survival rate in cancer patients. miR-937 overexpression promoted the viability, down-regulated the G1 phase ratios and increased the ability of colony formation in breast cancer cells. miR-937 inhibition inhibited the viability and the ability of colony formation, promoted the apoptosis and up-regulated the G1 phase ratios. Our results showed that miR-937 targeted bind to the APAF1-3'UTR. APAF1 overexpression inhibited the viability and the ability of colony formation, promoted the apoptosis and up-regulated the G1 phase ratios. After cells were co-transfection miR-937 mimics and APAF1, cell apoptosis level was increased. Conclusion: APAF1 up-regulation or APAF1 down-regulation in breast cancer may regulate cell proliferation and apoptosis.

FGFR4 provides the conduit to facilitate FGF19 signaling in breast cancer progression.

Although genetic amplification and overexpression of the fibroblast growth factor 19 (FGF19) gene are found in human breast cancer, mechanisms that contribute to such functional alterations remain elusive. We report here that high expression of FGF19 is associated with the aggressive malignant behavior and poor survival outcome of breast cancer patients. FGF19 is particularly highly expressed in luminal molecular subtype of breast tumors and its expression levels are positively associated with its secretion levels from breast cancer cells. Genetic knockout of FGF19 significantly induces repression of breast tumor progression and metastasis in either an orthotopic mouse model of breast cancer or an experimental metastasis model. The FGF19 specific receptor, FGFR4, can be activated and subsequently upregulate AKT signaling in breast cancer cell upon FGF19, which is critical for oncogenic role of FGF19. Inactivation of FGFR4 by its inhibitor BLU9931 significantly attenuates FGF19-induced tumor-promoting activity, suggesting interruption of FGFR4 function is sufficient to affect FGF19-driven breast cancer. Overall, these insights support the idea that targeting FGFR4 in breast cancer cells overexpressing FGF19 may represent an effective strategy to suppress cancer development, progression, and metastasis.

[A series of observation on the expression of TGF-beta1 in the lung of nitrofen-induced congenital diaphragmatic hernia rat model].

OBJECTIVE: To access the expression of transforming growth factor beta1 (TGF-beta1) in the lung of Nitrofen-induced congenital diaphragmatic hernia rat model. METHODS: Twelve timed-pregnant Sprague-Dawley rats were randomly divided into two groups, namely control group and CDH group on day 9.5 of gestation. Each rat in the CDH group was given 125 mg of Nitrofen (dissolved in seed fat) by gavage. Each rat in the control group was given the same dose of single oil. On day 16 of gestation, the two groups mentioned above were divided into three subgroups, and fetuses were delivered by cesarean section respectively on day 16, 18 and 21 of gestation. After the fetuses were checked for diaphragmatic hernia, lung tissue weight (LW) and body weight (BW) of each fetus on gestational day 21 were recorded. Lung histologic evaluations were made with microscope and TGF-beta1 immunohistochemistry staining were performed with image analyzing. RESULTS: At day 16 of gestation, no visible diaphragm closure was observed in all fetuses. Diaphragmatic hernia was observed in 32 of the 44 rat fetuses of the CDH groups on day 18 and day 21 of gestation (72.7%). Lw/Bw of the 21d subgroups of CDH group were lower than those of corresponding control group (P < 0.01). Observed under the microscope, the lungs of fetuses in CDH groups showed marked hypoplasia. The expression of TGF-beta1 was detected in cytoplasmic, without definite expression in nuclear. It was significantly stronger that the expression of TGF-beta1 was in the lungs of the CDH group than that of the control group (P < 0.01). CONCLUSIONS: Nitrofen interferes with lung development in early stage of the fetal before the diaphragm developed. TGF-beta1 would be one of the important factors which lead to pulmonary hypoplasia.

Effect of prenatal tetrandrine administration on transforming growth factor-beta1 level in the lung of nitrofen-induced congenital diaphragmatic hernia rat model.

PURPOSE: Tetrandrine (Tet) is a bisbenzylisoquinoline alkaloid isolated from the root of Stephania tetrandra, which has been used in traditional Chinese medicine to treat patients with silicosis, asthma, and pulmonary hypertension, and others and can be used as a pulmonary therapeutic agent. We hypothesized that it can also improve the lung growth in congenital diaphragmatic hernia (CDH) for its multiple biological effects. There are increasing evidences that suggest transforming growth factor beta1(TGF-beta1) plays a crucial role in fetal lung growth and morphogenesis. The aim of this study was to evaluate the effect of prenatal administration of Tet and to investigate its possible mechanism on the expression of TGF-beta1 in the lung of nitrofen-induced CDH rat model. METHODS: A CDH model was induced in pregnant Sprague-Dawley rats by administration of nitrofen on day 9.5 of gestation (Ed9.5 term, day 22). Tetrandrine (30 mg/kg) was given through gavage (once a day, for 3 days) on Ed11.5. Accordingly, there were 3 groups as follows: control (n = 9), CDH (n = 9), and CDH + Tet (n = 9). All the fetuses were delivered by cesarean delivery on Ed16.5, 18.5, and 21.5, respectively, to check if diaphragmatic hernia existed on each fetus, then the lung tissue weight (LW) and body weight (BW) of each fetus were recorded. Histologic evaluations and TGF-beta1 immunohistochemistry staining in the lung sample were performed for image analysis. RESULTS: Diaphragmatic hernia was observed in 95 of the 112 rat fetuses in CDH and CDH + Tet groups on Ed18.5 and Ed21.5 (84.8%), the incidence between the 2 groups had no statistical significance (P = .642). Lung weight/body weight in the CDH group and the CDH + Tet group were lower than that in the control group (P < .01), and LW/BW in the CDH group was lower than that in the CDH + Tet group (P < .05). Observed under the light microscope and electron microscope, marked hypoplasia of the lungs in fetuses among the CDH groups was observed, in contrast to improvement of the lungs in CDH + Tet fetuses. Statistical differences in morphological parameters (percentage of alveoli area, counting bronchus) were found even on Ed16.5 when diaphragm had not closed (P < .01). The number of type II pneumocytes and lamellar bodies in each group had no significant difference (P > .05). The immunoreactivity of TGF-beta1 in CDH group and CDH + Tet group were markedly stronger than that in the control group (P < .01). In addition, TGF-beta1 expression in the CDH group was stronger than that in the CDH + Tet group (P < .01). CONCLUSION: Nitrofen can interfere with lung development early in the fetal rat development before and separate from diaphragm development, and increased expression of TGF-beta1 in the lung of CDH rat model may suppress lung growth and development. Prenatal treatment with Tet can improve the growth of the lung of the nitrofen-induced CDH fetuses and its mechanism seems to be involved in downregulating the expression of TGF-beta1. It is a likely new approach to treat CDH and its coexistent lung hypoplasia by maternal Tet administration.

[Effect of tetrandrine on lung development: experiment of nitrofen-induced congenital diaphragmatic hernia fetal rat model].

OBJECTIVE: To assess the efficacy of prenatal administration of tetrandrine (TET) on pulmonary hypoplasia in the nitrofen-induced congenital diaphragmatic hernia (CDH) fetal rat model. METHODS: Six timed-pregnant female Sprague-Dawley rats were randomly divided into 3 equal groups: CDH group (receiving gavage of nitrofen 125 mg dissolved in seed fat on day 9.5), and TET group (receiving gavage of nitrofen 125 mg on day 9.5 and then gavage of TET 30 mg/kg on days 11.5 - 14.5), and control group (given the same dose of peanut oil on day 9.5 and the same dose of normal saline on days 11.5 - 14.5). The fetuses were delivered by cesarean section on day 21 to undergo light microscopy and electron microscopy. The numbers of type II pneumocytes were recorded and compared. RESULTS: CDH were detected in 32 of the 41 fetuses from the CDH and TET groups with a teratogenic rate of 78%, however, without a significant difference between the CDH and TET groups (P = 0.645). Microscopy showed significant lung hypoplasia in both histologic structure and cellular structure in the CDH group; however the lung development of the TET group was improved in comparison to the CDH group. There was no significant difference in numbers of type II pneumocytes among the 3 groups (P = 0.779). CONCLUSION: Prenatal administration of TET can improve the lung development of CDH rats in both histological structure and cellular structure. This may provide a new idea for the clinical treatment of CDH.

Effect of antenatal tetrandrine administration on endothelin-1 and epidermal growth factor levels in the lungs of rats with experimental diaphragmatic hernia.

PURPOSE: The aim of this study was to evaluate the effect of the traditional Chinese medicine tetrandrine (Tet) and to determine its possible mechanism on expression of endothelin-1 (ET-1) and epidermal growth factor (EGF) in the lung of a rat model of nitrofen-induced congenital diaphragmatic hernia (CDH). METHODS: A single oral dose (115 mg/kg) of nitrofen on day 9.5 of pregnancy was maternally administered to induce CDH. Pregnant rats were divided into 4 groups on day 18.5: control (n = 5), CDH (n = 5), CDH+dexamethasone (Dex) (n = 5), and CDH+Tet (n = 5). All fetuses were delivered by cesarean delivery on day 21.5. Accordingly, there were 4 groups of fetuses: control (n = 38), CDH (n = 25), CDH+Dex (n = 21), and CDH+Tet (n = 22). Lung tissue weight (LW) and body weight (BW) of each fetus were recorded, lung histologic evaluations and ET-1 and EGF immunohistochemistry staining were performed, and image analysis was performed after lung processing. RESULTS: Five female rats in the control group produced 38 fetuses without CDH. CDH was observed in 68 of the 128 rat fetuses (53.1%) among the other 3 groups. The LW/BW ratio of the CDH group was significantly lower than those of the Dex and EGF groups (P < .05). The lungs of fetuses with CDH showed marked abnormal structure such as pulmonary hypoplasia and vascular remodeling, in contrast to improved pulmonary structure in lungs of fetuses in the CDH+Dex and CDH+Tet groups. Statistical differences in morphologic parameters (radial alveolar counts, percentage of alveoli, percentage of medial wall thickness, and vascular volume) were found (P < .05). The immunoreactivity of EGF and ET-1 in the CDH group was markedly stronger than that in the control, CDH+Dex, and CDH+Tet groups (P < .01). In addition, EGF and ET-1 expression in the CDH+Dex and CDH+Tet groups was stronger than that in the control group (P < .05). There was no difference in lung EGF and ET-1 immunoreactivity between CDH+Dex and CDH+Tet groups (P > .05). CONCLUSION: Antenatal treatment with Tet may improve lung growth and vascular remodeling, and its mechanism seems to be involved in decreasing EGF and ET-1 expression. Tet administered maternally may be a hopeful new therapeutic option in the treatment of CDH and may be effective in helping to avoid the side effects of Dex.

A novel in vivo model of human hemangioma: xenograft of human hemangioma tissue on nude mice.

BACKGROUND: Experimental models of human infantile hemangiomas are needed, although none of the current ones is ideal in representing the natural development of hemangioma. In this article, the authors present a nude mice model of human hemangioma with serial morphologic findings on grafts. METHODS: A specimen of rapidly growing hemangioma tissue was obtained surgically from a 2-month-old boy and was cut into 96 pieces, which were implanted subcutaneously into nude mice, six pieces each. During the 6-month experimental period, gross view, hematoxylin and eosin staining, double immunofluorescent labeling with mouse antihuman CD31 and fluorescein isothiocyanate-labeled rat antimouse CD34, and transmission electron microscopic observation were performed. RESULTS: Eighty-one of the 96 grafts (84.4 percent) survived and grew during the second month to 3 to 4 months after transplantation, followed by gradual involution. Histologic examination showed cellular edema, degeneration, and necrosis in the early stage. From approximately 30 days, cellular density increased and karyokinesis was identified at 45 days. At 2 months, the grafts were composed mainly of hemangioma tissue and signs of involution appeared. Thereafter, signs of involution turned significant and the grafts were replaced with fibrofatty tissue. It was confirmed through double immunofluorescent labeling that the chief cellular components were endothelial cells of human origin. Findings on ultrastructural investigation were concordant with those on light microscopic observation. CONCLUSIONS: With human hemangioma tissue implanted into nude mice, an animal model was established successfully. The development of grafts after ischemia resembled the natural course of human infantile hemangioma.

[Effect of tetrandrine on endothelin expression in the lungs and clinical significance thereof: experiment with rat models with nitrofen-induced congenital diaphragmatic hernia].

OBJECTIVE: To investigate the effects of tetrandrine on endothelin expression in the lungs and its clinical significance. METHODS: 25 pregnant female SD rats were randomly divided into 5 equal groups: Groups A to D were fed with nitrofen to cause congenital diaphragmatic hernia (CDH) in the fetuses. Group A was injected with normal saline, Group B with dexamethasone (DXM), Group C with tetrandrine, and Group D with DXM + tetrandrine. Group E was control Group. On day 21.5 of pregnancy the fetuses were delivered by cesarean section and killed. Microscopy was used to observe the CDH formation, and the relative wall thickness (RWA) and relative wall area (RWA) of pulmonary arterioles. The lung/body weight ratio, and relative integrated optical density (IOD) of pulmonary arteriole and bronchiole were observed. The expression of endothelin in the lung tissues was detected by immunohistochemistry. RESULTS: 9 rats of Group A-D produced 57 fetuses with CDH with a CDH arte of 45.2%. The lung/body weight ratios, and RWA values of Group A-D were all significantly lower than that of Group E (all P < 0.05). The RWT of pulmonary arteriole was significantly lower in Groups B and C compared with Group E (both P < 0.05). The RWT and RWA of Group A were significantly lower than those of Group B-D (all P < 0.05). The values of relative IOD of pulmonary tissues and of pulmonary arteriole of Group A-D were all significantly lower than those of Group E (all P < 0.05). A positive correlation existed between the relative IOD of endothelin in pulmonary arteriole and in bronchiole (P < 0.01), and among the RWT and RWA of pulmonary arteriole, and relative IOD (all P < 0.01). CONCLUSION: Tetrandrine improves the pulmonary hypoplasia and degrades the pulmonary hypertension.