[Effect of Echinococcus granulosus hydatid cyst fluid protein on allergic rhinitis induced by ovalbumin in mice].

OBJECTIVE: To investigate the protective effect of Echinococcus granulosus hydatid cyst fluid protein (HCFP) on ovalbumin (OVA)-induced allergic rhinitis (AR) in mice. METHODS: Twenty-four BALB/c mice at ages of 8 to 10 weeks, each weighing approximately 20 g, were randomly divided into four groups, including groups A (blank control group), B (blank intervention group), C (AR model group) and D (AR+HCFP intervention group), with 6 mice in each group. On days 0, 2, 4, 6, 8, 10 and 12, mice in groups A, B, C and D were injected with 200 µL sterile phosphate buffered saline (PBS), 200 µL sterile PBS containing 20 µg HCFP, 200 µL sterile PBS containing 50 µg OVA and 5 mg Al(OH)3 gel, and 200 µL sterile PBS containing 50 µg OVA, 5 mg Al(OH)3 gel and 20 µg HCFP, respectively. On days 14 to 20, mice in groups A, B, C and D were administered with 40 µL sterile PBS, 40 µL sterile PBS containing 20 µg HCFP, 40 µL sterile PBS containing 2 mg OVA and 40 µL sterile PBS containing 2 mg OVA and 20 µL HCFP by nasal drop, respectively. Mouse behavioral changes were observed and behavioral scores were estimated. The serum levels of interferon-γ (IFN-γ), interleukin-4 (IL-4), IL-5, IL-10, transforming growth factor-ß (TGF-ß) and OVA-specific IgE antibody (OVA-sIgE) were measured using enzyme-linked immunosorbent assay (ELISA), and the pathological changes of mouse nasal mucosa were observed by hematoxylin and eosin (HE) staining. RESULTS: The mean behavioral score was significantly greater in Group C (6.83 ± 0.50) than in groups A (1.17 ± 0.52) and B (1.33 ± 0.52) (P < 0.05), while a lower mean behavioral score was estimated in Group D (3.50 ± 0.50) than in Group C (P < 0.05). There were significant differences among the groups in terms of serum IFN-γ (F = 4.08, P < 0.05), IL-4 (F = 275.90, P < 0.05), IL-5 (F = 96.82, P < 0.05), IL-10 (F = 77.67, P < 0.05), TGF-ß (F = 9.98, P < 0.05) and OVA-sIgE levels (F = 44.69, P < 0.05). The serum IFN-γ level was significantly lower in Group C than in groups A, B and C (P < 0.05), and the serum levels of IL-4, IL-5 and OVA-sIgE were significantly higher in Group C than in groups A, B and C (P < 0.05), while the serum IL-10 and TGF-ß levels were significantly greater in Group D than in Group C (P < 0.05). Microscopy showed apparent loss of nasal mucosa cilia, increased number and enlargement of goblet cells, interstitial edema and submucous vascular dilation in Group C, while the pathological changes of nasal mucosa were alleviated in Group D relative to Group C. CONCLUSIONS: E. granulosus HCFP has a protective activity against OVA-induced allergic rhinitis in mice.

Cisplatin suppresses tumor proliferation by inhibiting autophagy in ovarian cancer via long non-coding RNA RP11-135L22.1.

OBJECTIVE: To observe the effect of cisplatin-induced autophagy in human ovarian cancer cell lines and explore the correlation between RP11-135L22.1 with cisplatin-induced autophagy. MATERIALS AND METHODS: Genome-wide expression profile and chemotherapy sensitivity data of ovarian cancer were downloaded from TCGA database. It was found that the expression level of lncRNA RP11-135L22.1 differed between chemotherapy-sensitive group and insensitive group. Besides, RP11-135L22.1 expression levels were detected in 64 ovarian cancer tissues and 30 normal tissues by qRT-PCR. Relationship between RP11-135L22.1 expression levels in 64 ovarian cancer tissues and their clinicopathological characteristics were analyzed by x2-test. Cell viability was detected by CCK8 assay. Cell apoptosis and cell cycle were accessed by flow cytometry. HO8910 cells were selected for transfection of pcDNA-RP11-135L22.1, and qRT-PCR was used to evaluate RP11-135L22.1 expression in cisplatin-treated HO8910 cells. Western blot was performed to analyze the expression changes of autophagy-related proteins. RESULTS: Genome-wide expression profile of chemotherapy-sensitive and -insensitive patients with ovarian cancer from TCGA database was analyzed by edger package. It was found that RP11-135L22.1 level in chemotherapy-sensitive group was significantly lower than that of insensitive group. QRT-PCR results confirmed that RP11-135L22.1 was lowly expressed in ovarian cancer. The overall survival of patients was positively correlated with the expression of RP11-135L22.1. Furthermore, RP11-135L22.1 was associated with FIGO stage and tumor size. Flow cytometry showed that cisplatin could induce apoptosis and arrest cell cycle in ovarian cancer cells lines. CCK8 assay showed that cisplatin decreased viability of ovarian cancer cells. For in vitro study, HO8910 cells were cultured with medium containing different concentrations of cisplatin or treated with cisplatin for different times. The results revealed that RP11-135L22.1 expression was negatively correlated with the treating time and dose of cisplatin. Western blot showed that cisplatin induced autophagy in ovarian cancer cells in a time- and dose-dependent manner. Cisplatin combined with RP11-135L22.1 can reduce autophagy, increase the apoptosis and inhibit its activity of ovarian cancer cells to a certain extent. CONCLUSIONS: Cisplatin can induce autophagy in HO8910 ovarian cancer cells. After overexpression of RP11-135L22.1, it inhibited cisplatin-induced autophagy, thus enhancing the effect of cisplatin on ovarian cancer cells.

MicroRNA-574 enhances doxorubicin resistance through down-regulating SMAD4 in breast cancer cells.

OBJECTIVE: Drug resistance has become an important factor that threatens the survival and prognosis of patients with breast cancer, especially in patients with advanced breast cancer. Several microRNAs have been proved to participate in the resistant process; however, the role of miR-574 in doxorubicin (Dox) resistant breast cancer is still unclear. PATIENTS AND METHODS: Quantitative Real-time poly chain reaction (qRT-PCR) was employed to detect the expression level of miR-574 in breast cancer Dox-resistant MCF-7/Adr cell line and parental MCF-7 cell line. Using miR-574 mimics and inhibitors, miR-574 level was up- or down- regulated. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was handled to detect the IC50, and flow cytometric analysis was employed to measure the apoptosis and cell circle. Dual-luciferase and Western-blot experiments were applied to verify the direct target gene of miR-574. RESULTS: miR-574 expression level was significantly higher in MCF-7/Adr cells compared to normal MCF-7 cells. Up-regulation of miR-574 level in MCF-7 cells promoted the cell growth and G0/G1-to-S phase transition but inhibited cell apoptosis. However, knockdown of miR-574 in MCF-7/Adr cells decreased the IC50 and cell growth. Using luciferase assay, SMAD4 was confirmed to be a potential target of miR-574, and the expression of SMAD4 protein was regulated by miR-574. In blood samples of patients, the miR-574 level before chemotherapy was higher than that after chemotherapy. CONCLUSIONS: We revealed miR-574 could promote doxorubicin resistance of breast cancer MCF-7 cells via down-regulating SMAD4, thus providing a novel target for advancing breast cancer chemotherapy.

Ibrutinib presents antitumor activity in skin cancer and induces autophagy.

OBJECTIVE: Skin cancer is one of the most common malignancies in dermatology. Patient compliance and prognosis of skin cancer are poor. Ibrutinib, a Bruton's Tyrosine Kinase (BTK) inhibitor, is a new anticancer drug used to treat many cancers. Therefore, we aimed to explore the role of ibrutinib in the treatment of skin cancer. MATERIALS AND METHODS: Cell Counting Kit-8 (CCK8) and plate cloning assay were used to detect cell proliferation. Apoptosis was determined by flow cytometry. Western blotting analysis was used to analyze the expression of key proteins that regulated autophagy. Proliferation and apoptosis of skin cancer cells and induction of autophagy induced by ibrutinib were evaluated. RESULTS: CCK8 plate cloning assays showed that ibrutinib can gradually inhibit the skin cancer cell proliferation as the treatment time and dose increased. Results of flow cytometry showed that apoptosis in skin cancer cells were induced after ibrutinib treatment. Western blot showed that autophagy in skin cancer cells was found induced by ibrutinib and also related to the time and concentration of ibrutinib treatment. Combination treatment of ibrutinib and 3MA for skin cancer cells can significantly increase apoptosis. CONCLUSIONS: Ibrutinib has anti-tumor activity in skin cancer and can induce autophagy. Binding to autophagy inhibitors can promote ibrutinib's anti-skin cancer activity. Our experimental results provided new ideas for developing skin cancer drugs.

Circulating chemerin levels are increased in first-degree relatives of type 2 diabetic patients.

BACKGROUND: Chemerin is an important risk factor of insulin resistance and metabolic syndrome. The aim of this study was to explore the potential role of chemerin in the early stage of diabetes development. METHODS: 63 control subjects without any family history of diabetes and with normal glucose tolerance (NGT) and 74 healthy, first-degree relatives (FDRs) of type 2 diabetic patients were recruited in the study. All subjects underwent a 75 g oral glucose tolerance (OGTT) test after having fasted overnight. Plasma glucose, insulin, total cholesterol, HDL cholesterol, triglycerides, chemerin, and adiponectin were measured. RESULTS: FDR subjects had higher BMI, WHR, waist, fasting plasma glucose, fasting insulin, TG, UA, HOMA-IR, LDL-C, and lower HDL- C levels than control subjects (p < 0.05). The FDRs group had significantly lower adiponectin levels while chemerin was higher. Plasma chemerin levels were independently correlated with HOMA-IR, FINS, TG, FPG, and adiponectin level. Multiple stepwise regression analysis showed that HOMA-IR and TG were independent risk factors that influenced circulating chemerin levels. CONCLUSIONS: These findings showed a significant increase of chemerin levels in FDR subjects which suggested that chemerin may be involved in the development and progression of insulin resistance.