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Hsa-miR-145-5p kit and use of mature body analogy of Hsa-miR-145-5p

A technology of kit and mature body, applied in the application field of Hsa-miR-145-5p kit and its mature body mimic

Active Publication Date: 2014-03-26
FIRST HOSPITAL OF SHANXI MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, in the study of laryngeal squamous cell carcinoma, there are very few reports or related studies involving miRNAs

Method used

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  • Hsa-miR-145-5p kit and use of mature body analogy of Hsa-miR-145-5p
  • Hsa-miR-145-5p kit and use of mature body analogy of Hsa-miR-145-5p
  • Hsa-miR-145-5p kit and use of mature body analogy of Hsa-miR-145-5p

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] The kit consisting of Hsa-microRNA-145-5p and its target gene FSCN1 is used for the prognosis evaluation of laryngeal squamous cell carcinoma.

[0054] Specimen source and selection criteria

[0055] (1) Paraffin samples were obtained from patients with laryngeal squamous cell carcinoma who were first diagnosed and underwent surgery in the Department of Otorhinolaryngology, Head and Neck Surgery of the First Hospital of Shanxi Medical University from 2000 to 2006.

[0056] (2) The patient has not received radiotherapy, chemotherapy, or biological therapy before admission; except for the primary tumor in the larynx, other primary tumors have been ruled out by B-ultrasound, CT, MRI, PET and other examinations; the patient has no infections such as hepatitis B, syphilis, and tuberculosis Medical history, no family history of genetic diseases. The postoperative pathological diagnosis was squamous cell carcinoma of the larynx, and the mucosa of the resection margin adjace...

Embodiment 2

[0168] Example 2: Anti-tumor treatment of Hsa-miR-145-5p mature body mimics in laryngeal squamous cell carcinoma xenograft models established by Hep-2 and TU-177 in nude mice.

[0169] cell culture.

[0170] Laryngeal squamous cell carcinoma cell lines Hep-2 (China Center for Type Culture Collection, CCTCC), TU-177 (Bioleaf Biotech Co.), normal human oral mucosal keratinocytes HOK (SCIEN CELL). The culture condition of Hep-2 is RPMI-1640 (Hyclone) + 10% fetal bovine serum (Hyclone), and the culture condition of TU-177 is DMEM / F12 (Hyclone) + 10% fetal bovine serum (Hyclone). All cells were kept at 37°C, 5% CO 2 Cultured in a cell culture incubator. Passage with 0.25% trypsin solution containing EDTA.

[0171] Nude mice were bred.

[0172] BALB / C nude mice, purchased from Hunan SLK Jingda Experimental Animal Co., Ltd., SPF grade, female, 7 weeks old (certificate number: HNASLKJ20120391). SPF grade mouse feed was purchased from Guangdong Medical Experimental Animal Ce...

Embodiment 3

[0188] Example 3: Mimics of mature Hsa-miR-145-5p were used to inhibit the expression of markers of malignant mesenchymal transition, E-cadherin, N-cadherin, MMP-2 and MMP-9.

[0189] Experimental Materials

[0190] 1.1 Cell lines: human laryngeal squamous cell carcinoma cell lines Hep-2, TU-177.

[0191] 1.2 Cell culture reagents, methods and conditions are the same as before.

[0192] experimental method

[0193] 2.1 The day before transfection, the cells were 4 Inoculate each well on a 24-well plate, and the medium is DMEM containing 10% FBS

[0194] high sugar;

[0195] 2.2 On the day of transfection, the confluence of the cells is about 50-60%, aspirate the old medium, wash twice with PBS, and then

[0196] Add 300 μL OPTI-MEM (Invitrogen) medium, place in 5% CO 2 , 37 ° C incubator;

[0197] 2.3 Dilute 1 μL of Lipofectamine2000 (Invitrogen) with OPTI-MEM medium in each well, and the final volume is

[0198] 50 μL, let stand at room temperature for 5 minutes...

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Abstract

The invention discloses a Hsa-miR-145-5p kit and a use of a mature body analogy of the Hsa-miR-145-5p. The Hsa-miR-145-5p kit comprises the following quantitative detection components of a Hsa-miR-145-5p gene: a Hsa-miR-145-5p PCR primer, dNTP with the concentration of 10mM, an RNase inhibitor, a microRNA / cDNA reverse transcription primer, a buffer solution with the concentration of 5 times, M-MLV, a SYBRGreenPCRMasterMix mixing liquid with the concentration of 2 times, deionized water, a Hsa-miR-145-5pfluorescence in-situ hybridization probe and a FSCN1 immunohistochemical component, wherein the Hsa-miR-145-5pfluorescence in-situ hybridization probe is a base sequence complemented with a Hsa-miR-145-5p basic group and is modified by DNA skeleton and fluorophore cy3. The Hsa-miR-145-5p kit can be used for molecular targeting treatment medicine for Laryngeal carcinoma.

Description

[0001] technical field [0002] The invention relates to a human genus microRNA, in particular to the application of a Hsa-miR-145-5p kit and a mature body mimic thereof. Background technique [0003] Head and neck squamous cell carcinoma is the sixth most common tumor in the world and the fourth most common tumor among men in Europe. Laryngeal squamous cell carcinoma (LSCC) originates from the epithelium of the laryngeal mucosa, and its incidence rate ranks second among head and neck squamous cell carcinomas. bit. According to statistics, the incidence of laryngeal squamous cell carcinoma is increasing slightly all over the world, and the trend of rejuvenation is gradually appearing. [0004] The anatomical structure of the larynx is limited and submucosal lymphatic vessels are abundant, so laryngeal squamous cell carcinoma has the malignant biological characteristics of local invasion and cervical lymph node metastasis, which has become an important risk factor for recurre...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11G01N33/68
CPCC12Q1/6886C12Q2600/118C12Q2600/178G01N33/57407G01N33/57484G01N33/577G01N2333/47
Inventor 王斌全高伟张春明陈钢钢皇甫辉温树信冯彦
Owner FIRST HOSPITAL OF SHANXI MEDICAL UNIV
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