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Abstract Objectives Long non-coding RNAs (LncRNAs) act as an indispensable role in the Preeclampsia (PE)-related trophoblast function, while its relationship with Small Nucleolar RNA Host Gene 22 (SNHG22) remains unknown. Hence, this study aimed to investigate the roles of lncRNA SNHG22 in the Preeclampsia (PE)-related trophoblasts function and the underlying mechanism. Methods Normal placentas and placentas from PE patients were collected to detect the expression of lncRNA SNHG22. Then, trophoblasts HTR-8/Svneo and JEG-3 were purchased, cultured, and treated to investigate the roles of lncRNA SNHG22 on cell migration and invasion as well as its underlying regulatory mechanism. Results The SNHG22 was downregulated in PE patients, and it was found that SNHG22 overexpression could drive migration and invasion of trophoblasts, while SNHG22 depletion exerted a suppressive effect. Mechanistically, SNHG22 was validated to regulate microRNA-128-3p (miR-128-3p), and Protocadherin 11 X-Linked (PCDH11X) was identified as the target gene of miR-128-3p. Furthermore, it was found that SNHG22 acted as a promoter in the migration and invasion of trophoblast cells in a miR-128-3p/PCDH11X dependent manner, and SNHG22 silencing weakened the activation of PCDH11X-mediated PI3K/Akt signaling pathways through inhibiting miR-128-3p, thereby preventing migration and invasion of trophoblasts. Conclusion SNHG22 acted as a driver in the migration and invasion of trophoblasts and may be considered a candidate for the amelioration of PE.
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Objective@#To observe the migration of 4,4＇-diaminostilbene-2,2＇-disulfonc acid-based fluorescent whitening agents ( DSD-FWAs ) in food packaging paper, so as to provide evidence for quality and safety supervision for paper packaging materials.@*Methods@#Forty-one paper samples with DSD-FWAs positive were made into 6 cm2 pieces and were soaked in four food simulants ( distilled water, 3% acetic acid, 10% ethanol and 95% ethanol, 10 mL each ). The experiment was carried out at the specified soaking temperature and time. The migration amounts of eleven DSD-FWAs were detected by high performance liquid chromatography-fluorescence detection. @*Results@#C.I.220, C.I.24, C.I.210, C.I.85, C.I.113, C.I.264, C.I.353 and C.I.357 were found in all the four food simulants. At the same time and temperature, the migration amount was highest in 10% ethanol, followed by distilled water, 3% acetic acid and 95% ethanol. C.I.220 was dissolved in all four food simulants, in the range of 20-90 ℃, the migration amount increased with soaking temperature; at 20 ℃, 40 ℃ and 60 ℃, the migration amount increased first and then stabilized over time.@*Conclusion@#The higher the storage temperature and the longer the storage time of paper packaging, the easier the DSD-FWAs in packaging paper migrate to food.
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OBJECTIVES@#To investigate the expression and mechanism of the long non-coding RNA (lncRNA) HCG22 in oral squamous cell carcinoma (OSCC).@*METHODS@#HCG22 levels were detected in the OSCC and adjacent tissues, OSCC cells, and normal oral keratinocytes. HCG22 expression in SCC-25 and HSC-3 cells was upregulated by transfection of the overexpressing plasmi dvector. Methyl thiazolyl tetrazolium (MTT) assay, flow cytometry, and Transwell assay were employed to detect changes in cell proliferation, apoptosis, migration, and invasion ability, while Western blotting was used to detect the expression of epithelial-mesenchymal transformation-related proteins. The expression level of miR-650 in the cells was detected by real-time quantitative polymerase chain reaction (RT-qPCR), and dual-luciferase reporter gene assay was applied to assess the targeting relationship between HCG22 and miR-650.@*RESULTS@#Compared with that in adjacent tissues, the expression of HCG22 significantly decreased in OSCC tissues (@*CONCLUSIONS@#HCG22 is expressed at low levels in OSCC. Upregulation of the expression of this lncRNA can inhibit the proliferation, migration, invasion, and epithelial-mesenchymal transition of OSCC cells. The mechanism of action of HCG22 may be related to its targeted regulation of miR-650.
Subject(s)
Humans , Carcinoma, Squamous Cell/genetics , Cell Line, Tumor , Cell Movement , Cell Proliferation , Gene Expression Regulation, Neoplastic , Head and Neck Neoplasms , MicroRNAs/genetics , Mouth Neoplasms/genetics , RNA, Long Noncoding/genetics , Squamous Cell Carcinoma of Head and NeckABSTRACT
Objective To investigate the regulation of miR-939-5p on USP22 gene expression and its effect on liver cancer migration and proliferation.Methods The expression of miR-939-5p in hepatoma cell lines (HepG2,MHCC-97H,SMMC-7721,BEL-7404 and Huh7) and normal liver cell line LO2 was detected by realtime PCR (qPCR).The liver cancer cells with the lowest expression were used as experimental subjects,and transfected with miR-939-5p (Experimental group) or miR-NC (Control group).qPCR was used to detect the transfection efficiency of miR-939-5p.Transwell migration assay and MTT proliferation assay were used to detect the migration and proliferation of hepatoma cells after transfected miR-939-5p.Bioinformatics software predicted target genes for miR-939-5p.The dual luciferase reporter gene verified the interaction of miR-939-5p with the target gene.qPCR and Western blotting were used to detect the expression of target genes at mRNA and protein levels after over-expression of miR-939-5p.Measurement data were expressed as (Mean ± SD),and t test was used for comparison between groups.Results The expression of miR-939-5p was significantly lower in hepatoma cell lines than in normal hepatocytes (P ＜0.01),and the expression of miR-939-5p was the lowest in SMMC-7721 cells (P＜0.01).miR-939-5p was efficiently transfected into SMMC-7721 cells [(1.01 ±0.07) vs (20.12 ± 1.27),P ＜0.01].High expression of miR-939-5p inhibited the migration ability (P ＜ 0.01) and proliferative capacity of liver cancer SMMC-7721 cells (P ＜0.05).The USP22 gene may be a target gene of miR-939-5p.The luciferase reporter gene confirmed that miR-939-5p specifically binds to the 3'-UTR of USP22 mRNA (P ＜ 0.01).USP22 expression was decreased at mRNA and protein levels after high expression of miR-939-5p (P ＜ 0.01).Conclusions The expression of miR-939-5p was down-regulated in hepatocellular carcinoma cell lines,and miR-939-5p inhibited the migration and proliferation of liver cancer SMMC-7721 cells.The molecular mechanism was to interfere with the expression of USP22 gene.
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Objective To investigate the effect and underlying mechanism of micro ribonucleic acid (miR)-101 on the epithelial-mesenchymal transition in human hepatocellular carcinoma MHCC97H cells. Methods MHCC97H cell lines were transfected with miR-101 mimics (M101 group) and negative control mimic (NCM group), and the cell lines without transfection were used as the control group. The expression levels of miR-101 in cells of 3 groups were quantitatively measured using reverse transcription polymerase chain reaction (RT-PCR). Transwell assay was performed to evaluate the cell migration and invasion ability of 3 groups. The expression levels of vimentin, α-catenin, E-cadherin and USP22 proteins in cells of 3 groups were measured by Western blot. The relationship between miR-101 and USP22 was analyzed by dual-luciferase assay. Results In the M101 group, the expression level of miR-101 was significantly up-regulated, which was approximately 761 times of that in the control group (P<0.05). In the M101 group, the quantity of migrating cells was 15.7±1.6, significantly lower than 94.1± 1.8 in the control group (P<0.05). In the M101 group, the quantity of invasive cells was 9.1±0.4, significantly lower compared with 51.6±0.9 in the control group (P<0.05). In the M101 group, the expression levels of vimentin and USP22 protein were significantly down-regulated, whereas the expression levels of α-catenin and E-cadherin protein were significantly up-regulated. Dual-luciferase assay revealed that USP22 was the target gene of the downstream miR-101 signaling pathway. Conclusions miR-101 regulates the expression of epithelial-mesenchymal transition-related proteins and suppresses the epithelial-mesenchymal transition of hepatocellular carcinoma MHCC97H cells probably through down-regulating the expression level of USP22.
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Aim To investigate the up-regulation of miR-22 through Wnt pathway inhibits the proliferation,migration and invasion in human gastric MGC803 cells induced by diallyl disulfide(DADS).Methods The effects of proliferation,migration,and invasion of gastric cancer cells were evaluated by MTT,wound-healing and invasion assays.Online prediction software was applied to search the target gene of miR-22.Luciferase report gene assay was used to assess the target genes Wnt-1 of miR-22.The expressions of Wnt-1,β-catenin and TCF-4 were tested by qRT-PCR and Western blot,respectively.Results MTT showed that DADS and miR-22 notably decreased the proliferation compared with control group(P<0.05).Wound-healing assay showed that DADS and miR-22 could significantly inhibit the migration of MGC803 cells compared with the control group, especially in miR-22+DADS(P<0.05). Invasion assay showed that DADS and miR-22 could markedly inhibit the invasion of MGC803 cells compared with the control group, especially in miR-22+DADS(P<0.05). Online prediction software to search the target gene exhibited that Wnt-1 may be a target gene of miR-22. Luciferase report gene assay disclosed that Wnt-1 was identified as a direct target of miR-22. Qrt-PCR showed that the expression of Wnt-1 Mrna was respectively down-regulated by DADS and miR-22 compared withcontrol group, especially in miR-22+DADS(P<0.05). Western blot exhibited that DADS and miR-22 obviously suppressed the expressions of Wnt-1, β-catenin and TCF-4 proteins, especially in miR-22+DADS(P<0.05).Conclusion Up-regulation of miR-22 through Wnt pathway can remarkably suppress the proliferation, migration and invasion in MGC803 cells by DADS.
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AIM: To examine the expression of miRNA-22 in the ovarian tissues and the effect of miRNA-22 over-expression on the proliferation, migration and invasion in SKOV-3 cells.METHODS: The expression levels of miRNA-22 in different ovarian tissues and SKOV-3 cells were determined by qPCR .miRNA-22 was over-expressed by trans-fection of miRNA-22 mimic.The cell viability was examined by CCK-8 assay.The cell migration was measured by wound healing test .The cell invasion was analyzed by Transwell assay .The protein expression levels of VEGF and P 53 were deter-mined by Western blot .RESULTS: Compared with the normal ovarian tissue , the expression level of miRNA-22 was remarkably decreased in the ovarian tumor tissues .After transfection with miRNA-22 mimic, the expression level of miRNA-22 in the SKOV-3 cells was significantly increased , while the cell viability , migration and invasion were obviously decreased .Moreover , the protein expression of VEGF and P 53 was dramatically inhibited after over-expression of miRNA-22.CONCLUSION:The decreased miRNA-22 expression may be correlated with the development of ovarian can-cer.Over-expression of miRNA-22 decreases the cell viability , migration and invasion by reducing the protein expression of VEGF and P53.
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Background:The incidence of inflammatory bowel disease(IBD)is increasing recently. However,the pathogenesis has not been fully clarified. Aims:To investigate the expressions and significance of interleukin-22( IL-22),matrix metalloproteinase-9(MMP-9)and macrophage migration inhibitory factor(MIF)in peripheral blood of patients with IBD. Methods:A total of 80 patients with IBD admitted from May 2011 to Nov. 2014 at the First Affiliated Hospital of Soochow University were enrolled,in which 43 cases were Crohn’s disease(CD),37 cases were ulcerative colitis(UC). Forty healthy subjects were served as normal controls. Peripheral levels of IL-22,MMP-9 and MIF were detected by ELISA. Multivariate Logistic regression model was used to analyze IL-22,MMP-9 and MIF in active CD and UC and ROC curve was used to evaluate the diagnostic performance of these markers for screening of active CD and UC. Results:Compared with normal control group,peripheral levels of IL-22,MMP-9 and MIF increased significantly in CD and UC groups(P 0. 05). Peripheral levels of IL-22, MMP-9 and MIF in active CD and UC were significantly higher than those in remission stage(P < 0. 05). For screening of active IBD,the area under ROC curve(AUC)of combined detection of IL-22 and MMP-9(0. 853 for CD,0. 867 for UC) was superior to that of IL-22,MMP-9 or MIF only(0. 747,0. 770 and 0. 699 for CD,0. 774,0. 815 and 0. 761 for UC). Conclusions:Peripheral levels of IL-22,MMP-9 and MIF increase markedly in IBD patients,which are correlated closely with the activity of IBD. Combined detection of IL-22 and MMP-9 might greatly increase the accuracy for screening of active IBD.