Linc-ROR promotes invasion and metastasis of gastric cancer by activating epithelial-mesenchymal transition

Introduction: Previous studies have shown that the long intergenic non-protein coding RNA regulator of reprogramming (linc-ROR) is abnormally expressed in a variety of malignancies and plays an important role in tumor progression. However, little is known about the role of linc-ROR in gastric cancer. In this study, the relationship between the expression of linc-ROR and clinicopathological factors in gastric cancer and its potential mechanism were explored. Materials and Methods: The cells were classified into two groups: ROR small interfering RNA(si-ROR) and the Negative control siRNA (si-NC).Linc-ROR was knockdown in si-ROR group by small interfering RNA (siRNA). Detect the expression of linc-ROR in gastric cancer tissues and normal tissues and its relationship with clinicopathologic characteristics by RT-PCR. the invasion ability was studied by wound healing assay and transwell assay. The expression levels of EMT-related molecules was detected by RT-PCR and Western blotting. Result: Showed that the expression of lincROR in gastric cancer tissues was significantly higher than that in the adjacent normal tissues. The lincROR expression level was significantly related to the tumor grade, lymph node metastasis, and TNM stage in cancer tissues. The lincROR knockdown in gastric cancer cell lines significantly inhibited cell invasion and metastasis. It affected its malignant biological behavior by activating the epithelial-mesenchymal transition through increasing expression of vimentin as well as decreasing E-cadherin levels in gastric cancer cells. The lincROR silencing significantly decreased the expression of ?-catenin and c-myc. Conclusion: Linc-ROR can regulate cell invasion and metastasis by activating the epithelial-mesenchymal transition process partially through Wnt/?-catenin signal pathway in the gastric cancer cells. Link-ROR may be an important molecule for the metastasis of gastric cancer.

Up-regulated miR-452-5p suppressed the expression of RORα and promoted the proliferation and migration of HCC cells / 中国药科大学学报

@#To study the prognosis-related regulation mechanism of miR-452-5p and its influence on the proliferation and migration of hepatocellular carcinoma (HCC) cells, liver hepatocellular carcinoma (LIHC) dataset in The Cancer Genome Atlas (TCGA) was used to validate the differential expression of miR-452-5p and perform the Kaplan-Meier analysis of overall survival (OS).Target genes of miR-452-5p from TargetscanHuman and miRDB databases were predictived; and differentially expressed genes(DEGs) and weighted gene co-expression network analysis (WCGNA) were completed with GSE14520.Lipofectmine-2000 was used to transfect miR-452-5p mimics, mimics negative control, miR-452-5p inhibitor and inhibitor negative control into Huh7 cells,respectively.The mRNA and protein expression level of RORα in 4 groups were determined by RT-qPCR and Western blot.CCK-8 assay and Transwell assay were conducted to testify the capabilities of proliferation and migration.The regulation between miR-452-5p and RORα was confirmed by the dual luciferase reporter assay.After analysis in the TCGA-LIHC dataset, miR-452-5p had higher expression in HCC tissue than that in normal tissue, which was also associated with a shorter OS.RORα and LAMC1 were discriminated by intersecting of DEGs, WGCNA module genes, and predictive target genes.Survival analysis exhibited that dysregulation of RORα was significantly related to the OS.Overexpression of miR-452-5p in HCC cells suppressed the expression of RORα both in mRNA and protein, and also enhanced the viability and migration of HCC cells.The results of the dual luciferase reporter assay showed that miR-452-5p targeted 3′UTR of RORα.Up-regulated miR-452-5p inhibited the expression of RORα, facilitated the proliferation and migration of HCC cells, and promoted the progression and poor prognosis of HCC.

Effect of Warming and Heat-clearing Method on Expressions of FoxP3 and ROR-γt in Colon Tissue of Rats with Intestinal Endotoxemia Caused by Acute-on-chronic Liver Failure / 中国实验方剂学杂志

Objective::To study the effect of warming and heat-clearing method (Wenyang Jiedu Huayu decoction) on the expressions of Forkhead box P3 (FoxP3), Retinoic acid-related orphan receptor gamma t (ROR-γt) in colon tissue of mice with acute-on-chronic liver failure (ACLF), in order to explore the possible regulatory mechanism on intestinal endotoxemia (IETM) in liver failure mice. Method::The 130 SD rats were randomly divided into normal group (10 rats) and model group (120 rats). The ACLF mice model was established through the subcutaneous injection with bovine serum albumin and the intraperitoneal injection with D-galactosamine(D-Gal) and lipopolysaccharide (LPS). The model mice were randomly divided into model group, heat-clearing group (Yinchenhao decoction, 6.68 g·kg-1), warming group (Yinchen Zhufu decoction, 7.09 g·kg-1) and warming and heat-clearing group (Wenyang Jiedu Huayu decoction, 19.53 g·kg-1). The normal group and the model group were given distilled water by gastric lavage, while the other groups were given equal volume of corresponding Chinese herbal medicines for a week. The value of each index at 1, 12 and 24 h was measured. The ratio of Treg/Th17 cell in peripheral blood were detected and calculated by flow cytometry. Real-time fluorescence quantitative PCR (Real-time PCR) was used to detect the expressions of FoxP3 and ROR-γt in colon tissues of mice at different time points. In situ hybridization and immunohistochemistry were used to observe the expressions of FoxP3 and ROR-γt genes and proteins. Result::Compared with normal group, the ratio of Treg/Th17 in the model group decreased significantly at each time point (P<0.01). Compared with the model group, the Treg/Th17 ratio increased only in the warming and heat-clearing method group (P<0.05). Compared with normal group, the expression of ROR-γt in the model group was significantly higher (P<0.01), and the expression of ROR-γt in the model group was higher than FoxP3.Compared with the model group, the expressions of FoxP3 and ROR-γt mRNA in the heat-clearing group and the warming group decreased at each time point (P<0.05), and the expressions of FoxP3 and ROR-γt in the warming and heat-clearing method group decreased significantly (P<0.01). The expressions of FoxP3 and ROR-γt mRNA in warming and heat-clearing group decreased compared with those in the warming group and heat-clearing group (P<0.05). Conclusion::The mechanism of the warming and heat-clearing method on IETM in liver failure may be related to the regulation of FoxP3 and ROR-γt expressions.

Effect of Qi-Boosting Toxin-Resolving Granules and cisplatin on transplanted tumor of nasopharyngeal carcinoma and balance of Foxp3/ROR-γt / 中草药

Objective To investigate the immune tolerance of Qi-Boosting Toxin-Resolving Granules (QBTRF) to Treg in the micro- environment of nasopharyngeal carcinoma (NPC) transplanted tumor, and reveal the possible anti-tumor mechanism of traditional Chinese medicine. Methods Using tumor cells CNE2 to build grafted tumor models, the anti-tumor effect of the QBTRF and cisplatin was observed, and the protein expression levels of Foxp3, ROR-γt and the content of IFN-γ, TGF-β and IL-17 in serum were detected. Results It was found that the content of TGF-β and IL-17 in serum of the tumor granule group and the combination therapy group were significantly lower than the model group, the IFN-γ content was obviously higher than that of the model group and cisplatin group, the apoptotic rate was significantly higher than that of the model group, the protein expression of Foxp3 was significantly increased, and the expression of ROR-γt was obviously decreased, in which the combination group was obviously better than that of single-use groups (P < 0.01). Conclusion QBTRF can effectively regulate the balance of Treg/Th17 in mice, restore the function of Treg, enhance the immune function of T cells during chemotherapy in mice, and exert anticancer effect. Combined with cisplatin, it can reduce the toxicity of chemotherapy.

Expression and clinical significance of long non-coding RNA Linc00152, H19 and ROR in patients with esophageal cancer / 国际检验医学杂志

Objective To explore the diagnostic values of long non-coding RNA (lncRNA) Linc00152, H19and ROR in esophageal cancer, and to investigate the relationship between the three and the clinicopathological features of esophageal cancer.Methods Totally 185cases of esophageal cancer diagnosed (case group) in our hospital from October 2016to December 2017, and 200healthy controls (control group) were recruited during the same period.The levels of lncRNA Linc00152, H19and ROR in the peripheral blood of subjects were detected by real-time quantitative PCR (RT-qPCR).The diagnostic values of plasma lncRNA Linc00152, H19and ROR expression in esophageal cancer and the relationship between them and the clinical and pathological features of esophageal cancer were analyzed.Results The levels of plasma lncRNA Linc00152, H19and ROR in case group were higher than those in control group, and the difference were statistically significant (P＜0.05).When combined detection with plasma lncRNA Linc00152, H19, and ROR, the area under the curve (AUC) to distinguish esophageal cancer and healthy subjects was 0.977 (95％CI:0.902-0.998, P＜0.001), and the sensitivity was 90.6％, specificity was 83.3％.In addition, the expressions of lncRNA Linc00152, H19, and ROR in plasma of patients with esophageal cancer were significantly correlated with TNM stage, histology classification and differentiation degree of tumor (P＜0.05), but were not related to gender, age, and tumor sites (P＞0.05).Multivariate analysis showed that plasma lncRNA H19and ROR levels were both independent risk factors for esophageal cancer, which is of guiding significances for predicting the risk of esophageal cancer.Conclusion LncRNA Linc00152, H19and ROR have diagnostic values for esophageal cancer, and are significantly correlated with the differentiation degree, histology classification, and TNM stage of esophageal carcinoma.Plasma lncRNA H19and ROR have potential application value in predicting the risk of esophageal cancer.

Study on ADR Signal Detection of Dasatinib and Imatinib / 中国药房

OBJECTIVE:To mine the ADR signals of dasatinib and imatinib,and to provide reference for safe use of two drugs in clinic. METHODS:ROR and PRR method of disproportionality measures were used to mine the data in the reports about dasatinib and imatinib of 17 quarters from FDA ADE Reporting System during the fourth quarter of 2012-the fourth quarter of 2016. ADR description terms in reports were standardized with international medical terms dictionary. ADR with signal detected by both methods were screened again. RESULTS & CONCLUSIONS:Totally 505 ADR signals for dasatinib and 929 ADR signals for imatinib had been found by ROR and PRR. After re-screening,there were 351 ADR signals for dasatinib and 649 ADR signals for imatinib,including 153 ADR signals for both dasatinib and imatinib. ADR signals for both dasatinib and imatinib obtained in this study were in agreement with known safety information. ADR mainly occurred in gastrointestinal tract,blood and lymphatics,kidney and urinary system,cardiovascular and musculoskeletal tissue,etc. However,incomplete information in the instructions was also found,such as possible urinary system-related ADR signals caused by dasatinib were detected in this study is not mentioned in drug instruction;imatinib could cause ADR signals of left atrium and right atrium dilation,which were not included in their instructions. Common ADRs,such as headaches,metioned in drug instructions of imatinib,did not appear in the top 50 signal intensities in this study. In addition,the ADR of the two drugs also varied,such as 7 ADR signals as periorbital edema and ocular edoma of forimatinib were much stronger than dasatinib;5 ADR signals of dasatinib,such as pericardial effusion and pleural effusion were much stronger than imatinib,indicating clinical drug selection should be based on individual situation of patients.

The recent progress of Th17 and their effects in tumor immunity / 中国肿瘤生物治疗杂志

@#[摘 要] 人体免疫系统是由固有免疫和适应性免疫应答组成。适应性免疫应答在抗原入侵时扮演着至关重要的角色， 而CD4+ 辅助性T（CD4+ T helper, CD4+Th）细胞是适应性免疫应答的主要组成部分。最近新发现的一类不同于Th1和Th2细胞亚群且能 特征性分泌白细胞介素17 (interleukin-17, IL-17)的辅助性T细胞亚群，被命名为Th17细胞。Th17 细胞参与很多炎症性疾病、 自 身免疫性疾病和肿瘤等的发展过程，可通过分泌IL-17A、IL-17F、IL-21、IL-22、IL-23、粒细胞-巨噬细胞克隆刺激因子(granulocytemacrophage colony stimulating factor，GM-CSF)和干扰素γ（interferon-gamma，IFN-γ）等炎症细胞因子来发挥免疫效应和炎症效 应。但是Th17细胞是否参与肿瘤的发生发展、具体作用机制以及发挥促肿瘤还是抑肿瘤效应等问题存在很多争议。本文综述 了近年来Th17细胞分化调节过程的相关机制，以及其在肿瘤发生发展的作用，旨在为肿瘤的诊断和治疗提供新的思路。

The changes of ROR-γt,IL-17A and Foxp3 mRNA expression and promoter methylation in TDI-induced mouse asthma / 中国职业医学

OBJECTIVE: To explore the changes of retinoic acid related orphan receptor-γt( ROR-γt),interleukin( IL)-17 A and forkhead / winged helix protein 3( Foxp3) mRNA expression and promoter methylation in the process of asthma induced by toluene-diisocyanate( TDI). METHODS: Specific pathogens free grade healthy male BALB / c mice were randomly assigned into asthma group and control group with 18 animals in each group. In the asthma group,the mice were sensitized with 0. 30% TDI( mass-volume concentration) dropped on the dorsum of both ears( 20 μL / ear) on day 1 and day 8. On day 15,the mice were challenged with 20 μL 0. 01% TDI( mass-volume concentration) by the trachea. The control group mice were sensitized and challenged by the same procedures with the same amount of solvent( acetone / olive oil). The mice were challenged 24 hours,the pathological changes of trachea and lung tissues were observed. The bronchoalveolar lavage fluid( BALF) from each group was collected,and the inflammatory cells in BALF were counted and classified. IL-4and Interferon-γ( IFN-γ) levels in BALF supernatant were measured by enzyme-linked immunosorbent assay. ROR-γt,IL-17 A and Foxp3 mRNA expression in the lung tissue were measured by real-time fluorescent quantitative polymease chain reaction. The degree of ROR-γt,IL-17 A and Foxp3 promoter methylation in lung issue were detected by Mass Array system.RESULTS: The asthmatic group demonstrated the symptoms of acute asthma,such as breathing deeply and fastly,bowing the back,lifting the forelimbs,et al. But the control group had no such symptoms in mice. Hematoxylin-Eosin staining showed obvious inflammatory lesions in the trachea and lung tissue of asthmatic mice. Compared with the control group,the white blood cell count,the neutrophil and eosinophil percentages in BALF,the IL-4,IFN-γ levels in BALF supernatant in asthma group were all significantly increased( P < 0. 01),meanwhile the lymphocyte and monocyte percentages in BALF were reduced( P < 0. 01); ROR-γt mRNA expression was significantly increased( P < 0. 01),and the degree of promoter methylation from sites 3,4,5,6,8,11 and 12 was significantly reduced( P < 0. 05); IL-17 A mRNA expression was significantly increased( P < 0. 01),and the degree of promoter methylation from sites 6 and 7 was significantly reduced( P < 0. 01); Foxp3 mRNA expression was significantly reduced( P < 0. 01),and the degree of promoter methylation from sites 1 and 10 was significantly increased( P < 0. 01). CONCLUSION: Th17 / Treg cell immune imbalance occurs in asthma induced by TDI. ROR-γt,IL-17 A and Foxp3 gene promoter methylation abnormalities may be involved in Th17 / Treg cell immune imbalance.

<b>4. Safety Risk Evaluation Methodology in Detecting the Medicine Concomitant Use Risk which might Cause Critical Drug Rash</b> / 薬剤疫学

The new practical use example of the JADER datasets from Japanese Adverse Drug Event Report database opened by Independent Administrative Agency Pharmaceuticals and Medical Devices Agency in April, 2012 and afterwards will be reported. The purpose of this study is to examine the evaluating method of medicine concomitant use risk by the frequency at which two or more medicines were reported simultaneously, being assumed the possibility of the influence of drug interactions to be the concomitant use risk in an adverse drug event onset. In order to estimate the potential degree of the safety risk at the time of the concomitant use, the methodology was estimated by the following procedures. 1) For considering that two suspicion medicine targeted is one medicine, the statistical signal index which means those of medicines with use in the case where they both are indicated in one report, the index of the concomitant use, is computed. 2) The statistical signal index about two target suspicion medicines is computed individually. 3) The case where the ratio of the index of the concomitant use to the index obtained individually exceeds 2 also in any of two suspicion medicines is judged as there being the concomitant use risk. The Proportional Reporting Ratio (PRR) and the Reporting Odds Ratio (ROR) were used as a statistical signal index. In order to check the validity of this method, Stevens-Jonson Syndrome (SJS) and Toxic Epidermal Necrolysis (TEN) which are known for the adverse events of critical drug rash reported in JADER were taken up, and the causality of medical supplies limited to the medicine with which they were reported as a suspicion medicine. As for the combination of the suspicion medicine which fulfills the conditions of the concomitant use risk, 10 kinds of candidates out of 159 combinations for SJS and 22 kinds of candidates out of 111 combinations for TEN were detected, respectively. Although this approach for the concomitant use risk was considered to be an effective means in showing the above results, some issues about the ratio of the index of the concomitant use and criteria in the report numbers of the medicine to be chosen, the effective calculation method for combinations in more than 3 medicines, etc. will be required for the further examination.

<b>3. Evaluation of Drug Induced Severe Eruption Cases in the Japanese Adverse Drug Event Report Database and Commonality of the Reported Drugs</b> / 薬剤疫学

From April 2012, Japanese Adverse Drug Event Report database (JADER) has become downloadable for utilization in the public, under the specified acceptable use policy. Given the situation, we focused on the severe eruptions which cases are increased in the public Relief System for Sufferers from Adverse Drug Reactions, for the purpose to analyze the characteristics of typical severe eruptions and a trend or a commonality in the corresponding reported drugs, by utilizing JADER. Disproportionate reporting obtained with ROR (Reporting Odds Ratio) and distribution parameter estimations obtained with Weibull distribution fit for the onset time of drug adverse reactions, were applied for the analysis in addition to the summary of frequency. We obtained 10,171 cases of severe eruptions from JADER, after exclusion of duplicated reports. In the Drug Induced Hypersensitivity Syndrome (DIHS), which has characteristics in clinical time course and causal drugs, we confirmed that typical causal drugs such as anti-epilepsy are frequently reported in JADER. On the other hand, drugs other than typical causal drugs also showed high ROR signal values. In the estimation of Weibull distribution shape parameter fit for drug adverse reaction onset time, DIHS gave estimation apparently different from other severe eruptions. Coincide with the estimation, histogram of onset time for DIHS showed the peak at around 20 days after drug administration, which is later than other severe eruptions. We conclude that analytical approach to obtaining information from multiple aspects of JADER data should be a useful effort for the persons who are engaged in preventive action for drug adverse reactions.

Expression of ROR?t in pulmonary tissue of asthmatic mice and its relationship to airway inflammation / 中国免疫学杂志

Objective:To explore the expression of ROR?t in pulmonary tissue of asthmatic mice and to investigate the association between the expression of ROR?t and the airway inflammation.Methods:Thirty female BLAB/c mice were randomly divided into the control group,asthmatic group and dexamethasone (Dex)-treated group.The asthma model was induced by classical method with ovalbumin(OVA).The concentration of IL-17 in bronchoalveolar lavage fluid (BALF) and serum was measured by enzyme-linked immunosorbent assay(ELISA).Airway inflammation was evaluated by HE staining.The expressions of IL-17,ROR?t mRNA and protein was measured by reverse transcription-polymerase chain reaction(RT-PCR) and Western blot respectively.Results:The level of IL-17,ROR?t mRNA and protein of asthmatic group was significantly higher than those of control group and Dexamethasone treated group (P