Si hotair

Manufactured by GenePharma

Sourced in China

Si-HOTAIR is a laboratory equipment designed for the analysis and detection of the HOTAIR gene. It provides a platform for the quantification and characterization of HOTAIR expression levels in various biological samples. The core function of Si-HOTAIR is to enable researchers to study the role and expression patterns of the HOTAIR gene, which is known to be involved in various cellular processes and disease conditions.

Automatically generated - may contain errors

Lab products found in correlation

15 protocols using si hotair

Role of HOTAIR and miR-1 in ESCC

Check if the same lab product or an alternative is used in the 5 most similar protocols

Six ESCC cell lines (KYSE30, KYSE140, KYSE150, KYSE180, KYSE410, and KYSE510) were purchased from the German Culture Collection (DSMZ, Braunschweig, Germany). Human normal esophageal epithelium cells (HEEpiC) were obtained from ScienCell (Carlsbad, CA). These cells were grown in RPMI 1640 (Biowhittaker, Walkersville, MD) supplemented with 10% fetal bovine serum (HyClone, Logan, UT) at 37°C under 5% CO₂ and saturated moisture.
For investigating the role of HOTAIR, KYSE30 and KYSE510 cells were transfected with either siRNAs targeting HOTAIR (si-HOTAIR) or scrambled negative controls (si-NC; GenePharma, Shanghai, China). For investigating the relationship between HOTAIR and miR-1, HOTAIR cDNA and miR-1 mimics (GenePharma) were cloned into the mammalian expression vector pcDNA3.1 (Invitrogen, Carlsbad, CA) individually or in combination, and then the vectors were transfected into cells. For the function study of CCND1, KYSE30 and KYSE510 cells were transfected with either si-CCND1 or si-NC. All transfection reactions were performed by using Lipofectamine 2000 (Invitrogen) following the manufacturer's instructions.

Ren K., Li Y., Lu H., Li Z., Li Z., Wu K., Li Z, & Han X. (2016). Long Noncoding RNA HOTAIR Controls Cell Cycle by Functioning as a Competing Endogenous RNA in Esophageal Squamous Cell Carcinoma. Translational Oncology, 9(6), 489-497.

+ Open protocol

+ Expand

Transfection of HCC cells with miR-217 and HOTAIR modulators

Check if the same lab product or an alternative is used in the 5 most similar protocols

Si-HOTAIR, miR-217 mimics, the miR-217 inhibitor, and negative control were provided by GenePharma (Shanghai, China). A density of 2 × 105 Huh-7, Hep3B, SNU-387, and SNU-449 cells were plated into six-well plates and supplemented with 2 mL culture medium. The transfection of HCC cells was conducted using Lipofectamine™ 2000 (Invitrogen, Carlsbad, CA, USA) in accordance with the manufacturer's instructions. After 48 h cultivation, the cells were collected for subsequent experiments. The sequences were provided as followed:
Hsa-miR-217-5p mimics:
5'-UACUGCAUCAGGAACUGAUUGGA-3', 5'-CAAUCAGUUCCUGAUGCAGUAUU-3';
Hsa-miR-217-5p inhibitor:
5'-UCCAAUCAGUUCCUGAUGCAGUA-3';
HOTAIR-Homo-536:
Sense:5'-GCCUUCCUUAUAAGCUCGUTT-3'; Antisense:5'-ACGAGCUUAUAAGGAAGGCTT-3';
HOTAIR-Homo-1162:
Sense: 5'-CAAUAUAUCUGUUGGGCGUTT-3';
Antisense: 5'- ACGCCCAACAGAUAUAUUGTT-3';
HOTAIR-Homo-1597:
Sense: 5'- GGAAGCUCUUGAAGGUUCATT-3';
Antisense: 5'- UGAACCUUCAAGAGCUUCCTT -3';
Negative Control:
Sense:5'- UUCUCCGAACGUGUCACGUTT-3';
Antisense: 5'- ACGUGACACGUUCGGAGAATT-3';

Tang X., Zhang W., Ye Y., Li H., Cheng L., Zhang M., Zheng S, & Yu J. (2020). LncRNA HOTAIR Contributes to Sorafenib Resistance through Suppressing miR-217 in Hepatic Carcinoma. BioMed Research International, 2020, 9515071.

+ Open protocol

+ Expand

Investigating HOTAIR function via siRNA

Check if the same lab product or an alternative is used in the 5 most similar protocols

To investigate the function of HOTAIR, three types of small INTERFERing RNAs against HOTAIR (siHOTAIR) were synthesized by GenePharma Technologies (Shanghai, China). Transfection was performed with INTERFERin (Polyplus, France), and the efficiency of knockdown was examined by quantitative real-time PCR (qRT-PCR). The siHOTAIR with the highest knockdown efficiency was used for further study.

Li H., Cui Z., Lv X., Li J., Gao M., Yang Z., Bi Y., Zhang Z., Wang S., Li S., Zhou B, & Yin Z. (2020). Long Non-coding RNA HOTAIR Function as a Competing Endogenous RNA for miR-149-5p to Promote the Cell Growth, Migration, and Invasion in Non-small Cell Lung Cancer. Frontiers in Oncology, 10, 528520.

+ Open protocol

+ Expand

Hep-2 tumor xenograft model in NSG mice

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human CD34+ hematopoietic stem cell-engrafted NSG mice were obtained from the Jackson Laboratory (CD34+ hu-NSG™, Bar Harbor, ME, USA). All mice were maintained under specific pathogen-free conditions and adaptation for 2 weeks. 6 × 10⁶ Hep-2 cells were injected into the mice (n = 6) subcutaneously. Treatment started when tumor volume reached 80–150 mm³. Cholesterol-coupled si-NC or si-Hotair (GenePharma) combined with PBS or V1 were intratumorally injected into the tumor three times a week for two weeks. Tumors were measured using calipers every 3 d, and volumes (mm³) were calculated as follows: length × width²/2. 14 days after the last injection, mice were euthanized, and the tumors were carefully separated, followed by weighing and imaging.

Yuan X., Shen Q, & Ma W. (2022). Long Noncoding RNA Hotair Promotes the Progression and Immune Escape in Laryngeal Squamous Cell Carcinoma through MicroRNA-30a/GRP78/PD-L1 Axis. Journal of Immunology Research, 2022, 5141426.

+ Open protocol

+ Expand

Silencing HOTAIR Gene Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols

The small interfering RNA (siRNA) against HOTAIR (siHOTAIR) and non-targeting control siRNA (siNC) were purchased from Shanghai GenePharma Co., Ltd.. The sequences of the two siRNAs were as follows: siHOTAIR forward, 5′-AUUGAUUAGCUGUUUGUUCCC-3′ and reverse, 5′-AAAGUCUAGACAAUAGAUGGC-3′; siNC forward, 5′-CUAUUGUCUAGACUUUUAUCU-3′ and reverse, 5′-GAAAUCUGGUACAAAGGAAAG-3′. Cells were seeded into 6-well plates to 40–60% confluence and then transfected with siHOTAIR or siNC at a concentration of 60 nM using Lipofectamine 2000^® reagent (Invitrogen; Thermo Fisher Scientific, Inc.) and Opti-MEM I Reduced Serum Medium (Thermo Fisher Scientific, Inc.) following the manufacturer's protocol. At 36 h after transfection, cells were collected for subsequent experiments.

Sun X, & Chen Z. (2021). Cancer-associated fibroblast-derived CCL5 contributes to cisplatin resistance in A549 NSCLC cells partially through upregulation of lncRNA HOTAIR expression. Oncology Letters, 22(4), 696.

+ Open protocol

+ Expand

Overexpression and knockdown of miR-149-5p, HOTAIR, and DCLK1 in A549/DDP and H1299/DDP cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

For overexpression, miR-149-5p mimic and miR-NC mimic were purchased from GenePharma (Shanghai, China); the sequence of HOTAIR and coding domain sequence of DCLK1 were cloned into pcDNA3.1 (Invitrogen, Carlsbad, CA, USA), respectively. For knockdown, miR-149-5p inhibitor (in-miR-149-5p), siRNA against HOTAIR (si-HOTAIR), and their controls were obtained from GenePharma. Cell transfection was carried out with Lipofectamine™ 2000 (Invitrogen) according to the manufacturers’ instruction. After transfection for 24 h, A549/DDP and H1299/DDP cells were collected for further experiments. Sequences of siRNAs were as follows: si-HOTAIR: 5ʹ- GAACGGGAGUACAGAGAGAUU-3ʹ; si-NC: 5ʹ-GAACGGAGCGAGCAGACCUUU-3ʹ.

Zhan Y., Abuduwaili K., Wang X., Shen Y., Nuerlan S, & Liu C. (2020). Knockdown of Long Non-Coding RNA HOTAIR Suppresses Cisplatin Resistance, Cell Proliferation, Migration and Invasion of DDP-Resistant NSCLC Cells by Targeting miR-149-5p/Doublecortin-Like Kinase 1 Axis. Cancer Management and Research, 12, 7725-7737.

+ Open protocol

+ Expand

Regulation of HOTAIR and miR-488-5p in Cell Cultures

Check if the same lab product or an alternative is used in the 5 most similar protocols

The dulbecco’s modified Eagle’s medium (DMEM) was obtained from Gibco; Thermo Fisher Scientific, Inc. (Waltham, MA, USA). The 10% heat-inactivated fetal bovine serum (FBS) was achieved from Hyclone (Logan, USA) while the 1% antibiotic/antimycotic solution was from Sigma-Aldrich, Merck KGaA (Darmstadt, Germany). The lentivirus respectively containing HOTAIR, miR-488-5p, si-HOTAIR, si-miR-488-5p and negative control (NC) were purchased from Shanghai Gene Pharma Company (Shanghai, China). 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) was purchased from Sigma-Aldrich (St. Louis, MO, USA). The primary antibodies and horseradish peroxidase conjugated IgG were obtained from BD Bioscience (San Diego, CA, USA). The reagents used here were all analytical grade and obtained from Aladdin reagent co., Ltd. (Shanghai, China) if not mention elsewhere.

Xia F., Xia W, & Yu X. (2020). LncRNA HOTAIR Influences the Growth, Migration, and Invasion of Papillary Thyroid Carcinoma via Affection on the miR-488-5p/NUP205 Axis. Technology in Cancer Research & Treatment, 19, 1533033820962125.

+ Open protocol

+ Expand

Manipulation of HOTAIR and EZH2 in Chondrosarcoma

Check if the same lab product or an alternative is used in the 5 most similar protocols

SiEZH2, siHOTAIR and scrambled negative control siRNA (siNC) were synthesized in GenePharma (Suzhou, China). The sequences targeting HOTAIR and EZH2 are listed in Supplementary Table S1. The HOTAIR overexpression plasmid (pHOTAIR) was purchased from Addgene (Cambridge, MA, USA). MiRNA mimics and inhibitors were purchased from RiboBio (Guangzhou, China). RNAs were transfected into tumor cells using Lipofectamine3000 (Invitrogen, Carlsbad, CA, USA). MiR-454-3p and shHOTAIR stably expressed chondrosarcoma cells were infected with the lentivirus and selected with puromycin (1 μg/ml) for 4 weeks.

Bao X., Ren T., Huang Y., Sun K., Wang S., Liu K., Zheng B, & Guo W. (2017). Knockdown of long non-coding RNA HOTAIR increases miR-454-3p by targeting Stat3 and Atg12 to inhibit chondrosarcoma growth. Cell Death & Disease, 8(2), e2605-.

+ Open protocol

+ Expand

Regulation of GRP78 by HOTAIR in Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

The siHotair, siGRP78 and negative control (NC) were purchased from GenePharma (Shanghai, China). The Hotair overexpression plasmid (pHotair) was purchased from Addgene, while the plasmids pGRP78 and the control pVector were kindly provided by Dr. Ming-liang He (CUHK). Cells were seeded in 12-well plate and grew to 50% confluency for siRNA transfection and 90% for plasmid transfection. Plasmids and siRNAs were transfected with Lipofectamine 2000 (Invitrogen, USA) according to the manufacturer's protocol. At day 3 post-transfection, the cells were harvested and total RNA was extracted using TRIZOL reagent (Invitrogen, CA, USA). The total RNA was subjected to qRT-PCR analysis using the ImProm-II^™ Reverse Transcription System (Promega, USA) and SYBR^® Premix Ex Taq^™ II (TaKaRa). The primers for qRT-PCR were shown in Supplementary Table S2. All reactions were performed with a Step-One Plus Real Time PCR system (Applied Biosystems, USA) and done in triplicate.

Fu W.M., Lu Y.F., Hu B.G., Liang W.C., Zhu X., Yang H.D., Li G, & Zhang J.F. (2015). Long noncoding RNA hotair mediated angiogenesis in nasopharyngeal carcinoma by direct and indirect signaling pathways. Oncotarget, 7(4), 4712-4723.

+ Open protocol

+ Expand

Functional Regulation of miR-613 and HOTAIR

Check if the same lab product or an alternative is used in the 5 most similar protocols

The si‐HOTAIR, si‐NC, miR‐613 mimics, miR‐613 inhibitor and miR‐NC were all purchased from Genepharma (Shanghai, China). The recombinant plasmids of pcDNA3.1‐HOTAIR and pcDNA3.1/c‐Met were constructed with assistance of Invitrogen Life Technologies. The increased or decreased miR‐613 expression was achieved by transfection of miR‐613 mimics or miR‐613 inhibitors, while HOTAIR knockdown was achieved by transfecting si‐HOTAIR. Transfection was performed according to the instructions of Lipofectamine™ 2000 (Invitrogen), and each group of cells was harvested 24‐48 hours after transfection for further assays.

Yang G., Fu Y., Lu X., Wang M., Dong H, & Li Q. (2018). LncRNA HOTAIR/miR‐613/c‐met axis modulated epithelial‐mesenchymal transition of retinoblastoma cells. Journal of Cellular and Molecular Medicine, 22(10), 5083-5096.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!