Expression Level Of HOX Transcript Antisense RNA Research Articles

The diagnostic value of lncRNA HOTAIR for cervical carcinoma in vaginal discharge and serum.

There is a lower incidence of cervical carcinoma compared with other common carcinomas, however, the mortality rate of cervical carcinoma is higher, suggesting that the treatment and prognosis of cervical carcinoma are relatively poor. Therefore, cervical carcinoma patients urgently need to find new diagnostic markers for early detection and treatment. One hundred and fifty cervical carcinoma and 100 benign cervical disease patients from 2019 January to 2021 December in Tianjin Central Hospital of Gynecology Obstetrics were selected and 100 healthy women were as normal group. The expression of HOX transcript antisense RNA (HOTAIR) in cervical carcinoma and paracancerous tissue, serum sample was measured by realtime PCR assay. The receiver operating characteristic of HOTAIR for cervical carcinoma was analyzed. The study found that the expression level of HOTAIR in primary cervical carcinoma is closely related to tumor metastasis and prognosis. The expression level of HOTAIR in paracancerous tissue was significantly lower than that in cancer tissue, and the expression level of HOTAIR in vaginal discharge and serum was higher than that in cervical carcinoma patients which was positively correlated with tumor malignancy, meanwhile, HOTAIR was significantly reduced after surgery 3 months both in vaginal discharge and serum. In order to examine the diagnostic efficiency of HOTAIR for cervical carcinoma, we found that the area under curve of vaginal discharge was 0.9723, sensitivity was 92%, specificity was 98%, the area under curve of serum was 0.8518, sensitivity was 79%, and specificity was 94% by receiver operating characteristic analysis. The accuracy were 92.7% and 89.3% in vaginal discharge and serum via certified by cervical carcinoma and benign cervical disease patient and healthy people. The above results show that the diagnostic performance of HOTAIR in vaginal discharge is higher than that of serum, and it is expected to become a marker for cervical carcinoma diagnosis and treatment.

MiRNA-1246, HOTAIR, and IL-39 signature as potential diagnostic biomarkers in breast cancer.

The molecular alterations in noncoding RNA can lead to a cellular storm that is correlated to higher mortality and morbidity rates and contributes to the progression and metastasis of cancer. Herein, we aim to evaluate the expression levels and correlations of microRNA-1246 (miR-1246), HOX transcript antisense RNA (HOTAIR), and interleukin-39 (IL-39) in patients with breast cancer (BC). In this study, 130 participants were recruited, including 90 breast cancer patients and 40 healthy control participants. Serum levels of miR-1246 and HOTAIR expression were assessed by quantitative real-time polymerase chain reaction (qRT-PCR). Also, the level of IL-39 expression was evaluated using a Western blot. All BC participants demonstrated a remarkable elevation in miR-1246 and HOTAIR expression levels. Moreover, IL-39 expression levels demonstrated a noticeable decline in BC patients. Furthermore, the differential expression fold of miR-1246 and HOTAIR revealed a strong positive correlation among breast cancer patients. In addition, a negative relationship between the IL-39 and the miR-1246 and HOTAIR differential expression was also noticed. This study revealed that HOTAIR/miR-1246 exerts an oncogenic impact in patients with breast cancer. The expression levels of circulation miR-1246, HOTAIR, and IL-39 could be considered early diagnostic biomarkers in BC patients.

Investigation of LncRNAs expression in patients with hepatitis B virus

<b>Aim: </b>Patients infected with the hepatitis B virus (HBV) are at a higher risk of cirrhosis and hepatocellular carcinoma. Despite the recent advancement of antiviral therapy, many patients still cannot respond to existing therapies. Hence, to detect the changes in liver function earlier, non-invasive methods are needed. Long non-coding RNAs (lncRNAs) play important roles in essential biological process as well as human cancer. LncRNAs may be used as biomarkers in human diseases. Thus, in this study, we purposed to analyze the expression levels of lncRNAs (HOX transcript antisense RNA (HOTAIR), maternally expressed 3 (MEG-3), highly upregulated in liver cancer (HULC)) in patients with hepatitis B virus and healthy volunteers.<br /> <b>Methods: </b>We selected three lncRNAs as candidate lncRNAs based on their association with liver disease. Whole blood samples were collected from 40 patients with HBV and 48 healthy volunteers. The expression levels of all the samples were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR). Statistical analysis was implemented using GraphPad Prism software. A p-value lower than 0.05 was statistically meaningful.<br /> <b>Results: </b>The expression levels of HOTAIR and HULC were remarkably upregulated in the plasma of the patients with HBV compared with healthy control (p<0.05). In contrast, no significant difference in MEG-3 expression levels was observed between groups.<br /> <b>Conclusion: </b>Our findings showed that the expression of HOTAIR and HULC in plasma might be new promising diagnostic and/or prognostic biomarkers for HBV.

The Regulatory Effect of HOTAIR on the Protein and RNA Levels of ZEB1 and Its Potential Role in the EMT Process

Background: Invasion and metastasis in tumors are considered as two most important reasons for the reduction of treatment efficiency, as well as an increase in the mortality rate among the patients. According to the evidence, HOX transcript antisense RNA (HOTAIR) accounts for one of the main IncRNAs associated with the development and expansion of gastric cancer (GC). Objectives: This research investigates the impact of HOTAIR suppression on the expression and translation of the ZEB1 marker in GC. Methods: The knockdown HOTAIR was in the AGS cell line using small interfering RNA targeting against HOTAIR (si-HOTAIR). The impact of HOTAIR expression on the cell proliferation was previously evaluated by the MTT assay. The expression of the zinc finger E-box binding homeobox 1 (ZEB1) gene in transfected cell lines was quantified compared to the control samples using the quantitative real-time polymerase chain reaction (qRT-PCR). The enzyme-linked immunosorbent assay (ELISA) was also used to assess the HOTAIR suppression impact on the ZEB1 protein. Results: The findings revealed that a decrease in the HOTAIR expression could cause a reduction in the proliferation and growth of cancer cells (Fold changes = 0/28, P-value < 0.01). According to the impact of changes in the HOTAIR expression levels on the ZEB1 expression, the ZEB1 expression level was directly correlated with HOTAIR so that a decrease in the HOTAIR expression led to a significant decline in the ZEB1 expression level (P-value < 0.05). At the protein level, the effect of the knockdown of HOTAIR expression on the reduction of ZEB1 protein was also observed. Conclusions: Our findings showed a significant association between the HOTAIR and ZEB1 expression levels. Overall, the HOTAIR-ZEB1 axis plays a vital role in the epithelial-to-mesenchymal transition (EMT) process in human GC and represents a new therapeutic strategy for future GC treatment.

HOTAIR mediates cisplatin resistance in nasopharyngeal carcinoma by regulating miR-106a-5p/SOX4 axis

ABSTRACT This study explored the function and mechanisms of HOX transcript antisense RNA (HOTAIR) in the drug resistance of nasopharyngeal carcinoma (NPC). Quantitative PCR, Western blotting, MTT assay, flow cytometry, Transwell assay, and luciferase assay were performed. HOTAIR expression levels were upregulated in cisplatin (DDP)-resistant NPC tissues and cells. Knockdown of HOTAIR in DDP-resistant NPC cells increased cell sensitivity of DDP, as well as decreased cell viability, expression of chemoresistance-related proteins, migration and invasion, increased cell apoptosis. In addition, downregulation of microRNA 106a-5p (miR-106a-5p) expression and upregulation of SRY-box transcription factor 4 (SOX4) expression were observed in DDP-resistant NPC tissues and cells. MiR-106a-5p targets HOTAIR and SOX4; thus, silencing of HOTAIR significantly increased miR-106a-5p expression. The overexpression of miR-106a-5p significantly reversed the increase in SOX4 expression induced by HOTAIR lentivirus (Lv-HOTAIR). Knockdown of SOX4 reduced the drug resistance of DDP caused by the silencing of miR-106a-5p expression. In summary, HOTAIR enhanced DDP resistance in NPC cells by regulating the miR-106a-5p/SOX4 axis.

LncRNA HOTAIR facilitates high glucose-induced mesangial cell proliferation, fibrosis and oxidative stress in diabetic nephropathy via regulating miR-147a/WNT2B axis

BackgroundLong non-coding RNAs (lncRNAs) have been shown to be involved in the regulation of many disease progression. However, the role of lncRNA HOX transcript antisense RNA (HOTAIR) in diabetic nephropathy (DN) remains unclear.MethodsHigh glucose (HG)-induced human mesangial cells (HMC) was used to construct DN cell models in vitro. HMC proliferation was evaluated by CCK8 assay and EDU staining. Protein levels of proliferation markers, fibrosis markers, and wingless-type family member 2B (WNT2B) were measured using western blot analysis. HMC oxidative stress was assessed by determining the levels of oxygen species and malondialdehyde, as well as superoxide dismutase activity. Relative expression levels of lncRNA HOTAIR, microRNA (miR)-147a, and WNT2B were examined using quantitative real-time PCR. The interaction between miR-147a and lncRNA HOTAIR or WNT2B was confirmed by dual-luciferase reporter assay and RIP assay.ResultsOur data showed that lncRNA HOTAIR knockdown could inhibit the proliferation, fibrosis, and oxidative stress in HG-induced HMC. LncRNA HOTAIR could serve as a sponge of miR-147a. The inhibition effect of lncRNA HOTAIR silencing on the biological functions of HG-induced HMC could be reversed by miR-147a inhibitor. WNT2B was targeted by miR-147a, and its overexpression also overturned the suppressive effect of miR-147a on the proliferation, fibrosis, and oxidative stress of HG-induced HMC.ConclusionIn total, our research pointed out that lncRNA HOTAIR could mediate miR-147a/WNT2B axis to promote DN progression.

Effects of Long Noncoding RNA HOTAIR Targeting miR-138 on Inflammatory Response and Oxidative Stress in Rat Cardiomyocytes Induced by Hypoxia and Reoxygenation.

Objective To investigate the effects of HOX transcript antisense RNA (HOTAIR) and miR-138 on inflammatory response and oxidative stress (OS) induced by IRI in rat cardiomyocytes. Methods H9C2 cells were divided into the control group, H/R group, H/R+siRNA NC group, H/R+si-HOTAIR group, and H/R+si-HOTAIR+inhibitor group. Expression levels of HOTAIR, miR-138, and inflammatory factors were detected by quantitative reverse-transcription polymerase chain reaction (qRT-PCR). The double luciferase reporter gene assay was used to detect the targeting relationship between HOTAIR and miR-138. Results Compared with the control group, the level of miR-138 and SOD in the H/R group was obviously reduced, while the expression levels of the HOTAIR, MDA, and NF-κB pathway were obviously increased. Compared with the H/R group, the level of miR-138 and SOD in the H/R+si-HOTAIR group was obviously increased, and the expression levels of the HOTAIR, MDA, and NF-κB pathway were obviously decreased. Compared with the H/R+si-HOTAIR group, the level of SOD in the H/R+si-HOTAIR+inhibitor group decreased; MDA content and the NF-κB pathway expression level increased. In the double luciferase reporter gene assay, compared with the HOTAIR wt+NC group, the luciferase activity of the HOTAIR wt+miR-138 mimic group was obviously decreased. Conclusions Silent HOTAIR can promote the expression of miR-138 and inhibit H/R-induced inflammatory response and OS by regulating the NF-κB pathway, thus protecting cardiomyocytes.

HOTAIR Induces the Downregulation of miR-200 Family Members in Gastric Cancer Cell Lines

Background:Gastric cancer is the fourth most common human malignancy and the second reason for cancer morbidity worldwide. LncRNA HOTAIR has recently emerged as a promoter of metastasis in various cancer types, including GC, through the EMT process. However, the exact mechanism of HOTAIR in promoting EMT is unknown. Aberrant expression of the miR-200 family has been linked to the occurrence and development of various types of malignant tumors. This study investigates the correlation between the HOTAIR and miR-200 family gene expression patterns in GC cell lines. We investigated the miR-200 and HOTAIR due to their common molecular features in the EMT process. Methods:AGS and MKN45 cell lines were transfected with si-HOTAIR, along with a negative control. The effect of HOTAIR knockdown was also analyzed on cell viability and also on the expression of miR-200 family members, including miR-200a, -200b, and -200c, in cell lines using qRT-PCR. Statistical analysis was performed to find the potential correlation between the expression level of HOTAIR and miRs. Results:Our results showed significant increased miR-200 family expression level in transfected AGS and MKN45 GC cells (fold changes > 2; p < 0.001). Moreover, a negative correlation was observed between HOTAIR and miR-200 expression levels in GC cell lines (p < 0.05). Conclusion:Our findings showed a significant association between miR-200 family and HOTAIR expression levels in GC cell lines. Taken together, the HOTAIR-miR-200 axis seems to play a vital role in human GC, suggesting a potential therapeutic target in future GC treatment.

Silencing of HOTAIR Induced Apoptosis in Human Colorectal Cancer Cells through Up-regulation of Bax and Down-regulation of Bcl2

: HOX transcript antisense RNA (HOTAIR), as a long noncoding RNA (lncRNA) is a highly cited transcript modulating variety of signaling pathways such as cell growth and apoptosis. Altered expression of HOTAIR has been reported in human cancers, which contributes with cancer progression and metastasis. Increased expression level of HOTAIR has been observed in colorectal cancer (CRC). It seems that dysregulation of HOTAIR may inhibit the apoptosis. The present study was aimed to evaluate the effect of HOTAIR silencing on expression of apoptosis markers Bax and Bcl2 using real-time polymerase chain reaction (PCR). The data showed that HOTAIR and Bcl2 are highly expressed in CRC cells while the expression level of Bax is low. Following siRNA treatment, Blc2 was downregulated but Bcl2 was upregulated. These findings suggest that HOTAIR silencing can promote apoptosis, and thus it can be considered as a promising strategy to kill cancer cells.

Role and Mechanism of Exosome-Derived Long Noncoding RNA HOTAIR in Lung Cancer.

Background and purpose: HOX transcript antisense RNA (HOTAIR) is a long noncoding RNA (lncRNA) that promotes tumor growth and metastasis. Exosomes can mediate intracellular communication in cancer by transferring active molecules. However, the role and mechanism of HOTAIR in nonsmall cell lung cancer (NSCLC) are still unclear. This study mainly explores the role and mechanism of exosome-derived HOTAIR in NSCLC. Methods: after the material characterization of the CD63 immune lipid magnetic bead (CD63-IMB), the exosomes in serum of NSCLC patients were captured through CD63-IMB for the corresponding biological characterization. Real-time quantitative reverse transcription PCR (qRT-PCR) was performed to detect the expression level of HOTAIR in tumor tissues, serum, and serum exosome from NSCLC patients. Subsequently, exosome secreted by NCI-H1975 cells with highly expressed HOTAIR was selected to treat low-expression A549 cells and HOTAIR knockdown on NCI-H1975 cells. In this way, action mechanisms of HOTAIR can be investigated by means of qRT-PCR, colony formation assays, and flow cytometry. Results: exosomes can be isolated by CD63-IMB, and taken up by cells effectively; the qRT-PCR results demonstrate that HOTAIR expressions are significantly upregulated in tumor tissues, serums, and exosomes isolated from serums of NSCLC patients. Clinicopathological correlation analysis shows that the upregulation of HOTAIR is closely associated with lymphatic metastasis and tumor node metastasis (TNM) staging (P < 0.05). HOTAIR expressions show a significant increase in A549 cells treated with exosomes derived from NCI-H1975 cells, signifying that both proliferation and migration of A549 cells are promoted, and HOTAIR depletion could inhibit the proliferation and migration of lung cancer cells. Conclusions: HOTAIR is highly expressed in tumor tissues, serums, and serum exosomes of NSCLC patients and its expression has a significant correlation with lymphatic metastasis and TNM staging. Moreover, the exosome may promote NSCLC proliferation and migration through HOTAIR transportation. Therefore, exosome-derived HOTAIR is expected to be a new molecular marker for NSCLC diagnosis, and exosomal transmission of HOTAIR may provide a new approach to NSCLC diagnosis.

Long non-coding RNA HOTAIR knockdown alleviates gouty arthritis through miR-20b upregulation and NLRP3 downregulation

ABSTRACT This study aimed to determine the mechanism underlying the regulation of gout by the HOX transcript antisense RNA (HOTAIR) long non-coding RNA (lncRNA). The expression levels of HOTAIR, miR-20b, and Nlrp3 were estimated by qRT-PCR and western blotting. The methylation level of HOTAIR was detected by methylation-specific PCR. The recruitment of DNA methyltransferase 1 (DNMT1) to the lncRNA HOTAIR promoter was confirmed by a ChIP assay. RNA immunoprecipitation and RNA pull-down assays were used to confirm the interaction between HOTAIR and miR-20b. LncRNA HOTAIR and Nlrp3 expression was upregulated, and that of miR-20b was downregulated in synovial fluid mononuclear cells (SFMCs) collected from patients with gouty arthritis and monosodium urate (MSU)-stimulated THP-1 cells. Interleukin (IL)-1β level increased substantially upon stimulation by MSU crystals. The methylation percentage of HOTAIR was reduced in SFMCs from patients with gouty arthritis and MSU-stimulated THP-1 cells. DNMT1 expression was downregulated in MSU-stimulated THP-1 cells, and DNMT1 knockdown increased lncRNA HOTAIR expression. In addition, the interaction of HOTAIR with miR-20b was confirmed. HOTAIR knockdown suppressed Nlrp3 expression and the secretion of inflammatory cytokines via miR-20b regulation. Finally, in vivo experiments showed that HOTAIR knockdown alleviated ankle swelling in a mouse model of gouty arthritis. These findings suggest that lncRNA HOTAIR knockdown suppresses inflammatory cytokine secretion by upregulating miR-20b and downregulating NLRP3, thereby alleviating ankle swelling in gouty arthritis.

HOTAIR promotes paclitaxel resistance by regulating CHEK1 in ovarian cancer.

The HOX transcript antisense RNA (HOTAIR) has been reported to be aberrantly expressed in ovarian cancer (OC). Abnormal high expression level of HOTAIR has been found to be associated with poor overall survival of OC patients. Yet, the role of HOTAIR in paclitaxel resistance of OC is unclear. This study aims to investigate the effect, as well as the mechanism of HOTAIR in promoting paclitaxel resistance of OC. Ovarian cancer cell lines with down-regulated and up-regulated expression of HOTAIR were, respectively, established. The expression of HOTAIR was confirmed by qRT-PCR. The sensitivity of ovarian cancer cells to paclitaxel was detected by MTT assays, colony formation, EdU assays, flow cytometry, and in vivo experiments. An increased expression level of HOTAIR was observed in ovarian cancer cell lines following treatment with paclitaxel. When the expression of HOTAIR was down-regulated, the proliferation of ovarian cancer cells was found to be inhibited, coupled with enhanced cell sensitivity to paclitaxel. Conversely, when the HOTAIR expression was up-regulated, an opposite effect was observed on the ovarian cancer cells. In addition, cell cycle arrest in G2/M phase was also shown to be accelerated upon HOTAIR suppression. Strikingly, our results also revealed that HOTAIR plays a regulatory role in the expression of checkpoint kinase 1 (CHEK1), and that the restored paclitaxel sensitivity through knockdown of HOTAIR can be weakened by CHEK1 up-regulation. Consistently, in vivo data confirmed that the therapeutic efficacy of paclitaxel can be enhanced through down-regulation of HOTAIR, and that CHEK1 is the down-stream target of HOTAIR in inducing paclitaxel resistance. HOTAIR confers paclitaxel resistance in epithelial ovarian cancer by increasing the protein level of CHEK1.

Effect of lncRNA HOTAIR on the radiosensitivity of HCCLM3 cells

Objective: To investigate the effect of down-regulating long non-coding RNA (lncRNA) and HOX transcript antisense RNA (HOTAIR) targeting miR-761 on the radiosensitivity of HCCLM3. Methods: The expression of HOTAIR in liver cancer cells HuH-7, SNU-449, HCCLM3 and normal liver cells L-02 were measured by real-time quantitative PCR. HCCLM3 cells were divided into control, Sh-NC (transfected shRNA negative control), Sh-HOTAIR (transfected HOTAIR shRNA), RAD+Sh-NC (transfected shRNA negative control and irradiated with 8Gy dose), and RAD+Sh-HOTAIR (HOTAIR shRNA was transfected and irradiated with 8Gy dose) group. Apoptosis was detected by flow cytometry, Bcl-2 Associated X Protein (Bx-2), cleaved cysteine-containing Cleaved cysteinyl aspartate specific proteinase 3 (C-Caspase-3) protein expression. Sh-NC, Sh-HOTAIR cells were irradiated with 0, 2, 4, 6, 8 Gy, and plate-clone experiments were used to determine radiosensitivity. Bioinformatics software predicted that miR-761 might be a target gene of HOTAIR, and the luciferase reporter system identified the targeting relationship. The miR-761 inhibitor, HOTAIR shRNA and inhibitor negative control, and HOTAIR shRNA were co-transfected into HCCLM3 cells, respectively. Cell apoptosis and Bax and C-cysteine-containing aspartate proteins were also measured using the above method, as well as the hydrolase-3 protein expression and cell survival fraction. Results: The expression levels of HOTAIR in liver cancer cells HuH-7, SNU-449, and HCCLM3 were higher than those in normal liver cells L-02 (1.85±0.12, 2.27±0.23, 2.68±0.15 vs 1.00±0.09, P<0.05). Compared with Sh-NC, the apoptosis rate of Sh-HOTAIR, RAD+Sh-NC cells and Bax, C-Caspase-3 protein levels are higher [Apoptotic rate: (13.47±1.32)%, (12.84±1.19)% vs (2.98±0.27)%; Bax protein: 0.74±0.08, 0.72±0.06 vs 0.42±0.06; C-Caspase-3 protein: 0.56±0.06, 0.54±0.08 vs 0.25±0.04, all P<0.05]. Compared with Sh-HOTAIR and RAD+Sh-NC, RAD+Sh-HOTAIR cell apoptosis rate and Bax, C-Caspase-3 protein levels are higher [apoptosis rate:(22.57±2.36)% vs (13.47±1.32)%, (12.84±1.19)%, Bax protein: 0.99±0.11 vs 0.74±0.08, 0.72±0.06, C-Caspase-3 protein: 1.03±0.12 vs 0.56±0.06, 0.54±0.08,all P<0.05]. Compared with Sh-NC, Sh-HOTAIR cells had lower survival scores and higher radiosensitivity (P<0.05). HOTAIR targets negative regulation of miR-761 expression. Compared with cells co-transfected with inhibitor negative control and HOTAIR shRNA, cells co-transfected with miR-761 inhibitor and HOTAIR shRNA had lower apoptosis rate after radiation treatment [(10.24±1.32)% vs (21.84±2.01))%], Bax (0.50±0.06 vs 1.01±0.10) and C-Caspase-3 protein (0.56±0.07 vs 1.05±0.14) had lower expression and higher cell survival scores (all P<0.05). Conclusion: Down-regulating lncRNA HOTAIR targets miR-761 to increase the radiosensitivity of HCCLM3.

LncRNA HOTAIR enhances breast cancer radioresistance through facilitating HSPA1A expression via sequestering miR-449b-5p.

BackgroundBreast cancer (BRCA) is the leading cause of cancer‐related death in women worldwide. Pre‐ and postoperative radiotherapy play a pivotal role in BRCA treatment but its efficacy remains limited and plagued by the emergence of radiation resistance, which aggravates patient prognosis. The long noncoding RNA (lncRNA)‐implicated mechanisms underlying radiation resistance are rarely reported. The aim of this study was to determine whether lncRNA HOX transcript antisense RNA (HOTAIR) modulated the radiosensitivity of breast cancer through HSPA1A.MethodsA Gammacell 40 Exactor was used for irradiation treatment. Bioinformatic tools and luciferase reporter assay were adopted to explore gene expression profile and demonstrate the interactions between lncRNA, miRNA and target mRNA 3′‐untranslated region (3′‐UTR). The expression levels of certain genes were determined by real‐time PCR and western‐blot analyses. in vitro and in vivo functional assays were conducted by cell viability and tumorigenicity assays.ResultsThe levels of oncogenic lncRNA HOTAIR were positively correlated with the malignancy of BRCA but reversely correlated with the radiosensitivity of breast cancer cells. Moreover, the expression levels of HOTAIR were positively associated with those of heat shock protein family A (Hsp70) member 1A (HSPA1A) in clinical BRCA tissues and HOTAIR upregulated HSPA1A at the mRNA and protein levels in irradiated BRCA cells. Mechanistically, miR‐449b‐5p restrained HSPA1A expression through targeting the 3′‐UTR of HSPA1A mRNA, whereas HOTAIR acted as a competing sponge to sequester miR‐449b‐5p and thereby relieved the miR‐449b‐5p‐mediated HSPA1A repression. Functionally, HOTAIR conferred decreased radiosensitivity on BRCA cells, while miR‐449b‐5p overexpression or HSPA1A knockdown abrogated the HOTAIR‐enhanced BRCA growth under the irradiation exposure both in vitro and in vivo.ConclusionsLncRNA HOTAIR facilitates the expression of HSPA1A by sequestering miR‐449b‐5p post‐transcriptionally and thereby endows BRCA with radiation resistance.Key pointsTherapeutically, HOTAIR and HSPA1A may be employed as potential targets for BRCA radiotherapy. Our findings shed new light into the mechanism by which lncRNAs modulate the radiosensitivity of tumors.

Regulatory effect of microRNA-148b-3p on long non-coding RNA HOX transcript antisense RNA gene expression in human glioma cell line

Objective To investigate the regulation of HOX transcript antisense RNA (HOTAIR) gene by microRNA (miRNA, miR) in human glioma cell line. Methods Bioinformatics analysis was applied to predict the HOTAIR targeted miRNA. MiR-148b-3p mimics, miR-148b-3p inhibitor and their negative controls were bio-synthetized. Real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) was performed to analyze the regulation of HOTAIR gene expression by miR-148b-3p in human glioma cell line A172 after transferred with miR-148b-3p mimics. Lipofectamine 2000 transfecton and the Dual Luciferase Reporter Gene System were applied to detect the effect of miR-148b-3p on the HOTAIR gene in A172 cell. Results Results of RT-qPCR found that the expression level of HOTAIR in negative control group was 1.06±0.15. 24, 48 and 72 h after miR-148b-3p mimics transfection, the expression level of HOTAIR mRNA was 0.79±0.12, 0.63±0.18 and 0.39±0.06 respectively. 24, 48 and 72 h after miR-148b-3p inhibitor transfection, the expression level of HOTAIR mRNA was 1.24±0.09, 2.70±0.24 and 3.56±0.18 respectively. Compared with the negative control group, miR-148b-3p mimics could decrease the expression of HOTAIR gene (t=2.824, 3.102, 3.648, P 0.05); however, the luciferase activity of A172 cells co-transfected with mimics+ pmirGLO-WT-HOTAIR was significantly decreased compared with A172 cells co-transfected with miR-148b-3p mimics+ pmirGLO-MUT-HOTAIR (t=3.362, P<0.05). Conclusion MiR-148b-3p play a regulatory role of HOTAIR expression by directly targeted combination in human glioma cell lines. HOTAIR is a direct target of miR-148b-3p. Key words: Gliomas; Long non-coding RNA; HOX transcript antisense RNA; MicroRNA-148b-3p

Interaction Of c-Jun And HOTAIR- Increased Expression Of p21 Converge In Polyphyllin I-Inhibited Growth Of Human Lung Cancer Cells.

BackgroundLung cancer is a leading cause of cancer-related death worldwide. Previously we demonstrated that polyphyllin I (PPI), a bioactive component extracted from Paris polyphylla, inhibited the growth of non-small cell lung cancer (NSCLC) cells through the SAPK/JNK-mediated suppressing p65, DNMT1 and EZH2 expressions. However, the molecular mechanism underlying anti-lung cancer effect by PPI still remain elusive.PurposeIn this current study, we further explored the molecular mechanism underlying the anti-lung cancer effect of PPI.MethodsMTT, Cell-LightTM EdU DNA cell proliferation and colony formation assays were used to measure cell growth. Western blot were used to examine protein levels of c-Jun and p21. The expression level of long non-codingth RNA HOX transcript antisense RNA (HOTAIR) was measured by qRT-PCR. The p21 promoter activity was measured by Dual-Luciferase Reporter Assay System. The transient transfection experiments were used to silence and overexpression of c-Jun, p21 and HOTAIR. Tumor xenograft and bioluminescent imaging experiments were carried out to confirm the in vitro findings.Results We showed that PPI suppressed growth of NSCLC cells. Mechanistically, we observed that PPI reduced expression of HOTAIR, while increased transcription factor c-Jun protein levels. Additionally, PPI also induced protein expression and promoter activity of p21, a cyclin-dependent kinase inhibitor. While exogenously expressed HOTAIR showed no effect on c-Jun levels, silencing of c-Jun significantly reversed the PPI-inhibited HOTAIR expression. Moreover, excessive expressed c-Jun further enhanced PPI-inhibited HOTAIR expression and PPI-induced p21 protein levels. Intriguingly, overexpression of HOTAIR and silencing of c-Jun overcame the PPI-induced p21 protein and promoter activity. Finally, silencing of p21 neutralized the PPI-inhibited cell proliferation. Similar results were also found in one xenograft mouse model.Conclusion Our results demonstrate that PPI inhibits growth of NSCLC cells through regulation of HOTAIR and c-Jun expressions, which lead to induction of p21 gene. The interactions among HOTAIR, c-Jun and p21 regulatory axis converge in the overall anti-lung cancer effect of PPI. This study unveils an additional new mechanism for the anti-lung cancer role of PPI.

Deregulated Expression of Long Non-coding RNA HOX Transcript Antisense RNA (HOTAIR) in Egyptian Patients with Multiple Myeloma.

Increasing evidence of involvement of non-coding RNAs, especially long non-coding RNAs (lncRNAs), in the molecular biology of various malignancies have been recently reported. Their utilization as markers for diagnosis, prognosis and evaluation of treatment response was widely investigated. As the impact of lncRNA HOTAIR on multiple myeloma (MM) was not properly highlighted, we aimed to explore the expression levels of HOTAIR in three groups of MM patients and to analyze its relationship to different patients' characteristics. Plasma samples were withdrawn from 24 newly diagnosed MM patients, 23 post-therapy patients in complete response (CR) or very good partial response (VGPR) and 15 patients who had either progressive disease (PD) or relapse. The expression of lncRNA HOTAIR in MM patients and 20 healthy controls was analyzed by quantitative reverse transcription polymerase chain reactions. HOTAIR was significantly upregulated in newly diagnosed and PD/relapse categories in comparison with controls and MM patients who had achieved CR or VGPR (P < 0.001). Furthermore; HOTAIR expression levels correlated with the percentage of malignant plasma cells in bone marrow (P = 0.006) and disease stage (ISS stage) (P = 0.031). HOTAIR may be employed as prognostic molecular marker and novel therapeutic tool for newly diagnosed MM patients.

LncRNA HOTAIR inhibited osteogenic differentiation of BMSCs by regulating Wnt/β-catenin pathway.

This study aims to investigate whether HOX transcript antisense RNA (HOTAIR) can participate in the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) by regulating the Wnt/β-catenin pathway, thereby participating in the pathogenesis of osteoporosis. We detected the expression level of HOTAIR in 60 osteoporosis patients and 60 normal controls by Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR). Meanwhile, BMSCs derived from human or rats were subjected to determination of HOTAIR level. Subsequently, the effects of HOTAIR on osteogenic differentiation were evaluated by the activity of Alkaline Phosphatase (ALP), Alizarin Red S (ARS) staining, ALP staining and osteogenic-specific gene expression. The expression level of proteins related to the Wnt/β-catenin was determined by Western blot, and ALP activity was detected by ALP activity determination kit and alizarin red staining after knockdown or overexpression of HOTAIR, as well as the treatment of DKK1 or the Wnt pathway antagonist. Finally, osteoporosis model in rats was established by ovariectomy (OVX). We examined protein levels of HOTAIR, β-catenin, CyclinD, C-myc, and Runx2 in rat bone tissues. Bone morphology was observed in each group as well. The serum and BMSCs levels of HOTAIR in patients with osteoporosis were remarkably higher than that in normal people. Inhibition of HOTAIR induced increased ALP activity increased osteogenic marker genes and enhanced number of calcified nodules in BMSCs. However, the overexpression of HOTAIR exhibited the opposite effects. HOTAIR inhibited the expression level of Wnt/β-catenin pathway-related protein. Also, Wnt pathway antagonist DKK1 partially reversed the regulatory effects of HOTAIR on Wnt/β-catenin. DKK1 treatment markedly reduced the promotive effect of HOTAIR knockdown on ALP activity, ALP content and calcification ability of BMSCs. DKK1 administration in rats undergoing OVX showed worse bone morphology relative to controls. Protein levels of HOTAIR, β-catenin, CyclinD, C-myc and Runx2 remarkably downregulated in OVX rats administrated with DKK1. HOTAIR inhibits osteoblast differentiation of rat BMSCs. The underlying mechanism of which may be related to the mediation of Wnt/β-catenin pathway.

Gene Expression of the Circulating Long Noncoding RNA H19 and HOTAIR in Egyptian Colorectal Cancer Patients.

Background: HOX transcript antisense RNA (HOTAIR) and H19 are two long noncoding RNAs that play vital functions in the development of colorectal cancer (CRC). Subjects and Methods: The expression level of HOTAIR and H19 in the sera samples of Egyptian CRC patients along with normal controls was evaluated by quantitative real-time PCR. The possible correlations with the biochemical and clinicopathological characteristics were determined. Results: The expression of HOTAIR and H19 showed 7.55- and 11.38-fold increased levels, respectively, in CRC patients compared to the controls (p < 0.001). Furthermore, HOTAIR expression in CRC patients with regional lymph node metastasis was significantly higher when compared with CRC patients without regional lymph node metastasis (p = 0.034). HOTAIR and H19 expression showed no significant correlation with tumor site or carcinoembryonic antigen concentration. The sensitivity and specificity of HOTAIR and H19 in the detection of CRC cases were calculated as 92.9% and 100%, respectively. Conclusion: HOTAIR and H19 expression levels are upregulated in Egyptian CRC patients, and therefore can be considered noninvasive diagnostic biomarkers with high sensitivity and specificity.

Long non-coding RNA DRHC inhibits the proliferation of cancer cells in triple negative breast cancer by downregulating long non-coding RNA HOTAIR.

Long non-coding RNA (lncRNA) downregulated in hepatocellular carcinoma (DRHC) is a tumor suppressor in liver cancer. However, the role of this lncRNA in breast cancer has not been investigated. The present study revealed that lncRNA DRHC was downregulated and lncRNA Hox transcript antisense RNA (HOTAIR) was upregulated in tumor tissues compared with adjacent healthy tissues in patients with triple negative breast cancer (TNBC). Expression levels of lncRNA DRHC and lncRNA HOTAIR were negatively correlated in tumor tissues but not in adjacent healthy tissues. The lncRNA DRHC expression level was correlated with tumor size but not tumor metastasis. In vitro overexpression of lncRNA DRHC in TNBC cell lines resulted in decreased expression of lncRNA HOTAIR; however, lncRNA HOTAIR overexpression did not affect the expression level of lncRNA DRHC. Overexpression of lncRNA DRHC inhibited, while overexpression of lncRNA HOTAIR promoted the proliferation of the TNBC cell lines. In addition, lncRNA HOTAIR overexpression attenuated the inhibitory effects of lncRNA DRHC overexpression on cancer cell proliferation. The results obtained in the current study suggested that lncRNA DRHC may inhibit the proliferation of TNBC cells by downregulating the expression of lncRNA HOTAIR.