In situ hybridization probe, reagent and application of long non-coding RNA LOC401317

A long-chain non-coding and in-situ hybridization technology, which is applied in the field of in-situ hybridization probes of long-chain non-coding RNA LOC401317, can solve the problems of unknown lncRNA function and other problems, and achieve the effect of important promotion and application prospects and far-reaching clinical significance.

Active Publication Date: 2015-03-04
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, more than 40,000 human lncRNAs have been cloned, but the functions of most lncRNAs are unknown

Method used

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  • In situ hybridization probe, reagent and application of long non-coding RNA LOC401317
  • In situ hybridization probe, reagent and application of long non-coding RNA LOC401317
  • In situ hybridization probe, reagent and application of long non-coding RNA LOC401317

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1, the TP53 gene was introduced into nasopharyngeal carcinoma cells, and the expression of lncRNA LOC401317 was significantly upregulated

[0043] 1. Materials and methods:

[0044] 1.1 Reagents and kits

[0045] Common biochemical reagents such as agarose (agrose) and gel recovery kits were purchased from Shanghai Huashun Biological Engineering Co., Ltd. Mini Kit (Qiagen) extraction kit extracts RNA with improved quality, SuperScript TM The III (Invitrogen) kit reverse transcribed RNA into cDNA. The Luciferase Reporter Assay Kit (Dual-Luciferase Reporter Assay System) was purchased from Promega. The nasopharyngeal carcinoma cells HNE2 used in the present invention are preserved by the Cancer Institute of Central South University. The RPMI1640 medium and fetal bovine serum used for cell culture, and the trypsin used for digesting cells are all products of Gibco, USA. The lncRNA chip is a product of Agilent, the chip size is 4*180K, and there are 46506 lnc...

Embodiment 2

[0076] Example 2, lncRNA LOC401317 inhibits nasopharyngeal carcinoma cell cycle arrest and induces apoptosis

[0077] 1. Materials and methods

[0078] 1.1 Reagents and kits

[0079] Restriction enzymes Nhe I and EcoR I and T4DNA ligase were purchased from TakaRa Company; TRIZOL TM Reagent (Invitrogen); Plasmid Extraction Kit, Gel Recovery Kit (OMEGA); Reverse Transcription Kit (Promega); Proteinase K, DNase I, RNAsin, RNase A (GBICOL Company); Tetramethylazolazolium Blue ( MTT, Sigma); antibiotic G418 (Ameresc).

[0080] 1.2 Construction of pcDNA3.1-LOC401317 eukaryotic expression vector

[0081] We chose pcDNA3.1 blank vector (from Invitrogen Company) to construct the overexpression vector of LOC401317. We chose Nhe I and EcoR I restriction sites for digestion of pcDNA3.1 vector and inserted LOC401317 sequence into this site.

[0082]The steps to construct pcDNA3.1-LOC401317 eukaryotic vector are as follows:

[0083] 1) Using the cDNA of HNE2 cells transfected with TP...

Embodiment 3

[0133] Example 3, LOC401317 inhibits the growth of nasopharyngeal carcinoma cells in a nude mouse transplantation tumor model

[0134] 1. Materials and methods

[0135]Thirty male BALB / C nude mice, 4 weeks old, weighing 19±2g, were purchased from Shanghai Slack Experimental Animal Co., Ltd. All nude mice passed the quality inspection and were bred under specific pathogen-free (SPF) conditions in the Experimental Animal Department of Central South University.

[0136] The construction of LOC401317 eukaryotic expression vector and the preparation, cell culture and transfection of polylysine-modified silicon nanoparticles are the same as in Example 2.

[0137] Take 2×10 HNE2 cells transfected with LOC401317 or pcDNA3.1 blank vector, and HNE2 cells (Mock) without any treatment 6 Inject into the subcutaneous of the armpits of nude mice respectively, observe and compare the growth of the tumors, measure the size of the transplanted tumors with a vernier caliper across the skin fro...

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Abstract

The invention discloses an in situ hybridization probe, a reagent and an application of long non-coding RNA LOC401317. The long non-coding RNA LOC401317 can be used for preparing a prognosis preparation for a patient with nasopharyngeal carcinoma, and particularly a kit for predicting prognosis of the patient with the nasopharyngeal carcinoma employing an in situ hybridization detection method is prepared. A research proves that expression of the LOC401317 in a nasopharyngeal carcinoma tissue is lowered, and the patient with the nasopharyngeal carcinoma in low-expression LOC401317 is worse than the patient with the nasopharyngeal carcinoma in high-expression LOC401317 in prognosis, therefore, expression of the LOC401317 is applied to prognosis prediction of the patient with nasopharyngeal carcinoma; a powerful molecular biology basis can be provided for prognosis of the patient with the nasopharyngeal carcinoma; and the long non-coding RNA LOC401317 has profound clinical significance and important popularization and application prospects.

Description

technical field [0001] The invention belongs to the field of tumor molecular biology, and in particular relates to an in-situ hybridization probe, reagent and application method of long-chain non-coding RNA LOC401317. Background technique [0002] Nasopharyngeal carcinoma is a common malignant tumor of the head and neck with a high incidence, prone to cervical lymph node metastasis and poor prognosis. Studies have shown that the occurrence and development of this tumor is a complex process involving multiple genes, multiple steps, and multiple stages, in which the inactivation of tumor suppressor genes and the activation of oncogenes play a key role. [0003] Since the TP53 gene (encoding p53 protein) was cloned in 1979, it has been one of the most striking and important tumor suppressor genes in the field of tumor molecular biology research. A large amount of evidence shows that TP53 gene is widely involved in the occurrence and development of various malignant tumors by r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6886C12Q2600/118C12Q2600/158C12Q2600/178
Inventor 李桂源曾朝阳熊炜龚朝建李小玲张文玲李夏雨曾勇周鸣马健彭淑平向波
Owner CENT SOUTH UNIV
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