Preliminary study on the mechanism of dexmedetomidine regulating lncRNA HOTAIR to improve lung injury in septic mice / 中华急诊医学杂志

Objective:To investigate the mechanism of dexmetomidine (DEX) in improving lung injury in septic mice.Methods:Male C57BL/6 mice were randomly assigned to the blank group (NC), sham operation group (sham), cecal ligation and puncture group (CLP), and Dex treatment group (CLP+DEX), 36 mice per group. Mice in the CLP group were intraperitoneally injected with 1 mL sterile saline 15 min before CLP, and mice in the CLP + DEX group were intraperitoneally injected with 50 μg/kg DEX 15 min before CLP. The survival rate was recorded within 24 h after CLP. The mice were sacrificed at 0, 3, 6, 12, and 24 h after CLP, and lung tissues were collected. The expression levels of cytokines (IL-6, IL-1β, TNF-α) and lncRNA-HOTAIR in the lung of mice were detected by qPCR. RAW264.7 cell were cultured in vitro, LPS (100 ng/mL) and DEX (1 μ mol/L) were used to establish a cell model for studying the mechanism of Dex, and the expression of cytokines (IL-6, IL-1β, TNF-α) and lncRNA-HOTAIR in RAW264.7 cell model were detected by qPCR. In addition, the effect of lncRNA-HOTAIR on sepsis was explored in vivo and in vitro by knockdown or overexpression of HOTAIR.Results:The survival rate of the CLP+DEX group was higher than that of the CLP group within 24 h after surgery, and the levels of IL-6, IL-1β, and TNF-α in the lungs were significantly lower than those in the CLP group at 6, 12, and 24 h after surgery ( P<0.05). In addition, the level of lncRNA HOTAIR showed that the expression level of lncRNA HOTAIR in the lungs of mice were decreased after Dex treatment, and were decreased 1.1 times ( P<0.05), 4.0 times ( P<0.01) and 3.8 times ( P<0.01) at 6, 12, and 24 h, respectively. Compared with the NC group, knockdown of HOTAIR significantly decreased the levels of IL-1β, IL-6, and TNF-α in septic mice ( P<0.05), and overexpression of HOTAIR significantly increased the levels of IL-1β, IL-6, and TNF-α in septic mice ( P<0.01). Conclusions:DEX can reduce the production of inflammatory factors in the lungs of septic mice and improve the survival rate of septic mice. The mechanism may be related to the inhibition of HOTAIR expression.

Efficacy of Jianpi Xiaoai Prescription in Inhibition of Metastasis of Colon Cancer HCT116 Cells： An Exploration Based on LncRNA HOTAIR/JAK2/STAT3 Signaling Pathway / 中国实验方剂学杂志

Objective:To observe the effect of Jianpi Xiaoai prescription on long non-coding RNA Hox transcript antisense intergenic RNA （lncRNA HOTAIR）/Janus kinase 2 （JAK2） /signal transducer and activator of transcription 3 （STAT3） signaling pathway and to explore the potential mechanism of Jianpi Xiaoai prescription in suppressing the metastasis of colon cancer. Method:The expression of lncRNA HOTAIR in different cells was analyzed. Following the treatment of HCT116 cells with 10%，15%，and 20% Jianpi Xiaoai prescription -containing serum， the invasive ability of Jianpi Xiaoai prescription on HCT116 cells was assessed by transwell assay. The mRNA expression levels of lncRNA HOTAIR，JAK2，and STAT3 were measured by Real-time polymerase chain reaction （Real-time PCR）， and the protein expression levels of JAK2， phosphorylated STAT3 （p-STAT3） and STAT3 by Western blot. Result:The highest expression of lncRNA HOTAIR was detected in HCT116 cells. Compared with the blank group， each Jianpi Xiaoai prescription group exhibited a decreased number of invasive cells （<italic>P</italic><0.05， <italic>P</italic><0.01）. The relative JAK2 mRNA expression in the middle-dose Jianpi Xiaoai prescription group was down-regulated （<italic>P</italic><0.05）， and the relative lncRNA HOTAIR mRNA expression in the middle- and high-dose Jianpi Xiaoai prescription groups and the relative JAK2 mRNA expression in the high-dose Jianpi Xiaoai prescription group were remarkably down-regulated （<italic>P</italic><0.01）. Compared with the blank group，the relative p-STAT3 protein expression was down-regulated in the middle-dose Jianpi Xiaoai prescription group （<italic>P</italic><0.05）， and the relative JAK2 protein expression in the middle- and high-dose Jianpi Xiaoai prescription groups and the relative p-STAT3 protein expression in the high-dose Jianpi Xiaoai prescription group were remarkably down-regulated （<italic>P</italic><0.01）. Conclusion:Jianpi Xiaoai prescription effectively inhibits the metastasis of colon cancer cells， which may be related to the inhibition of lncRNA HOTAIR/JAK2/STAT3 signaling pathway.

Construction and screening of fibroblast-like synoviocytes in human rheumatoid arthritis stably overexpressing lncRNA HOTAIR / 医学研究生学报

Objective　The HOTAIR gene is closely related to pannus formation in rheumatoid arthritis (RA). This study aimed to construct and screen fibroblast-like synoviocytes in human RA (HFLS-RA) stably overexpressing lncRNA HOTAIR, and to pave the way for further study of the role of lncRNA HOTAIR in the pathogenesis of RA.　Methods　LncRNA HOTAIR was cloned and linked to the PMT406 vector digested by BamHI-HF-HF and XhoI. The constructed plasmids were sequenced, identified and then transfected into 293T cells to pack lentivirus. The HFLS-RA cells were infected with the recombinant and empty vector lentiviruses, and purinomycin was employed to screen the lncRNA HOTAIR-overexpressed and control cell lines. The total RNA was extracted from the blank, negatively transfected and overexpressed cells by Trizol, and the cDNA obtained by reverse transcription was amplified by qPCR, followed by determination of the expression of lncRNA HOTAIR.　Results　The relative expression of lncRNA HOTAIR was significantly higher in the overexpression group than in the blank control and negative transfection groups (30.329 ± 3.860 vs 1.001 ± 0.048 and 0.892 ± 0.247, P 0.05).　Conclusion　The HFLS-RA cell line stably overexpressing lncRNA HOTAIR was successfully constructed, which has provided some experimental evidence for further investigation of the role of lncRNA HOTAIR in the pathogenesis of RA.

Effect of silencing of long non-coding RNA HOTAIR on radiosensitivity of rectal adenocarcinoma cell lines / 中华放射肿瘤学杂志

Objective To investigate the effect of long non-coding RNA HOX transcript antisense RNA ( lncRNA HOTAIR ) on the cell proliferation, radiosensitivity and apoptosis of the rectal adenocarcinoma cell lines SW480 and HCT116 in vitro. Methods The expression levels of lncRNA HOTAIR in the rectal adenocarcinoma cell lines ( SW480 and HCT116 ) were assessed by real-time quantitative PCR (RT-qPCR).Silencing of HOTAIR by RNA interference was performed to explore its roles in cell proliferation,radiosensitivity and apoptosis. After treatment with irradiation at a gradient dose,the cell viability was measured and the rate of cell apoptosis was tested. Results Compared with the human rectal epithelial cell lines,the expression of lncRNA HOTAIR was significantly higher in the rectal adenocarcinoma cell lines. The colonic assay demonstrated that the sensitizing enhancement ratios ( SERs) were 1. 58 and 1. 33 for the cells transfected with HOTAIR siRNA in SW480 and HCT116 cell line compared with the control isRNA transfection group. In vitro silencing of lncRNA HOTAIR could enhance the apotosis rate and radiosensitivity of the rectal adenocarcinoma cell line SW480. Conclusion The expression level of lncRNA HOTAIR is correlated with the cellular radiosensitivity, which is probably a parameter for predicting the radiosensitivity of rectal adenocarcinoma cells. Radiotherapy combined with HOTAIR-siRNA can significantly inhibit the cell proliferation,induce cell apoptosis and enhance the radiosensitivity.

Influence of long chain non-coding RNA (lncRNA)-HOTAIR on migration and invasion ability of endometrial cancer / 临床与实验病理学杂志

Purpose To investigate the influence of long chain non-coding RNA (lncRNA)-HOTAIR on endometrial cancer cell proliferation,invasion,metastasis and other biological behaviour.Methods 20 cases of endometrial carcinoma tissue specimen,20 cases of hyperplasia tissue sample and 10 cases of normal tissue specimen were collected.Difference expression of lncRNA-HOTAIR in normal endometrium,hyperplasia endometrium tissue and endometrial carcinoma tissue at all periods were detected with RT-PCR assay.HOTAIR-siRNA transfection into Ishikawa cells was utilized with Lipofectamine 2000.MTT experiment was used to detected the proliferation ability of cells in all groups.Transwell chamber experiment was used to test the migration and invasion ability of cells in all groups.Results The gene expression level of of lncRNA-HOTAIR in endometrial carcinoma group at all stages was prominently increased compared with normal endometrium group (P ＜ 0.05).The expression level of lncRNA-HOTAIR in simple hyperplasia endometrium group and atypical hyperplasia endometrial group was not significantly different (P ＞ 0.05).Cell proliferation,invasion ability and migration ability of HOTAIR-siRNA targeting suppression group were lower than the blank control group and the negative control group significantly (P ＜ 0.05).Conclusion lncRNA-HOTAIR may involved in occurrence and development of endometrial cancer,which may play an important role in the aggression and metastasis of endometrial cancer.