Application of kidney cancer prognosis novel molecular marker non-coding RNA LINC00158, kit and detection method

A molecular marker, non-coding technology, applied in the field of tumor molecular biology, to achieve the effect of improving postoperative life quality, improving survival rate, and far-reaching clinical significance

Inactive Publication Date: 2018-11-02
THE FIRST AFFILIATED HOSPITAL OF ZHENGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Whether LINC00158 can be used as a new tumor marker to pre...

Method used

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  • Application of kidney cancer prognosis novel molecular marker non-coding RNA LINC00158, kit and detection method
  • Application of kidney cancer prognosis novel molecular marker non-coding RNA LINC00158, kit and detection method
  • Application of kidney cancer prognosis novel molecular marker non-coding RNA LINC00158, kit and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1: The application of non-coding RNA LINC00158, a novel molecular marker for prognosis of kidney cancer, in the preparation of preparations for predicting the prognosis of kidney cancer patients.

Embodiment 2

[0021] Example 2: Preparation of reagents for detecting the expression of non-coding RNA LINC00158 for the preparation of a kit for the prognosis of renal cancer patients (for 30 reactions) 1. Trizo 120ml

[0022] 2. Inhibit RNA degradation solvent 40ml;

[0023] 3. Chloroform 80ml;

[0024] 4. 80ml of isopropanol;

[0025] 5. DEPC water 10ml;

[0026] 6. 100 μl of a mixture of 10× random primers and Oligo dT primers;

[0027] 7. 150 μl of 5× reverse transcription reaction buffer;

[0028] 8. 100μl of 10mM deoxyribonucleotide triphosphate base dNTP containing Mg2+;

[0029] 9. 200U / μl M-MLV reverse transcriptase 50μl;

[0030] 10. SYBR Green qPCR Mix 500μl;

[0031] 11. 3 μM target gene LINC00158 specific primer (its sequence is shown in SEQ NO:2 and SEQ NO:3) 50 μl;

[0032] 12. 50 μl of 3 μM internal reference gene TUBA1A-specific primer (its sequence is shown in SEQ NO:4 and SEQ NO:5).

Embodiment 3

[0033] Example 3: Detection of non-coding RNA LINC00158 in tissue samples

[0034] 1. Collect the kidney cancer or normal control tissues to be tested, wash them with normal saline, put them into a cryopreservation tube filled with a solvent that inhibits RNA degradation, and put them in a -80°C refrigerator for later use.

[0035] 2. Extraction of RNA in tissues:

[0036] (1) Add liquid nitrogen to the mortar first, then cut the tissue into small pieces and grind it into powder in liquid nitrogen, take 100mg of tissue powder with a liquid nitrogen pre-cooled spoon and add it to the EP tube filled with 1ml of Trizol solution middle. The total volume of tissue powder should not exceed 10% of the volume of Trizol used, and it should be mixed thoroughly;

[0037] (2) Leave at room temperature for 5 minutes, then add 200 μL of chloroform, tightly cap the EP tube and shake vigorously for 0.5 minutes. Centrifuge at 12000 rpm for 10 minutes;

[0038] (3) Take the upper aqueous ph...

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Abstract

The invention relates to application of kidney cancer prognosis novel molecular marker non-coding RNA LINC00158, a kit and a detection method. Through real-time fluorescent quantitative PCR and survival curve analysis, kidney cancer tissue derived non-coding RNA LINC00158 is related to the survival rate of patient; the higher is content is, the higher the survival rate is; and the method providespowerful technical support for analyzing the prognostic survival period of the patients suffering from kidney cancer, is favorable for improving the postoperative life quality of the patients suffering from kidney cancer, formulating postoperative treatment scheme and increasing the survival rate, and has profound clinical significance.

Description

technical field [0001] The invention belongs to the field of tumor molecular biology, and in particular relates to an application, a kit and a detection method of a novel molecular marker non-coding RNA LINC00158 for prognosis of kidney cancer. Background technique [0002] Renal cell carcinoma is a common tumor of the urinary system. With the gradual westernization of people's lifestyles, the incidence rate is increasing year by year. Among them, the most common pathological type of renal cell carcinoma is clear cell carcinoma, accounting for about 70% of the total, followed by Papillary carcinoma, and then chromophobe cell carcinoma, often occurs in middle-aged and elderly people, and the incidence rate of young people is also increasing year by year. The incidence ratio of male to female is about 2:1. Since there are no symptoms in the early stage, about 20%~ 30% of patients have metastasized at the time of first diagnosis, and the prognosis of early stage patients is bet...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12Q1/6851
CPCC12Q1/6851C12Q1/6886C12Q2600/118C12Q2600/158C12Q2600/178C12Q2531/113C12Q2545/101C12Q2563/107C12Q2561/113
Inventor 李兆明
Owner THE FIRST AFFILIATED HOSPITAL OF ZHENGZHOU UNIV
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