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Pancreatic cancer prognosis molecular marker non-coding RNA Lnc-DCTD-25 and application thereof

A molecular marker, rnalnc-dctd-25 technology, applied in the field of tumor molecular biology, to achieve the effect of prolonging survival time, huge economic and social benefits, and improving survival rate

Inactive Publication Date: 2018-09-04
THE FIRST AFFILIATED HOSPITAL OF ZHENGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Whether Lnc-DCTD-25 can be used as a new tumor marker preparation reagent and kit for predicting the prognosis of patients with pancreatic cancer has not been published yet.

Method used

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  • Pancreatic cancer prognosis molecular marker non-coding RNA Lnc-DCTD-25 and application thereof
  • Pancreatic cancer prognosis molecular marker non-coding RNA Lnc-DCTD-25 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Preparation of reagents for detecting the expression of Lnc-DCTD-25 used to prepare a kit for the prognosis of patients with pancreatic cancer (for 30 reactions)

[0033] 1. Trizo l20ml

[0034] 2. Inhibit RNA degradation solvent 40ml

[0035] 3. Chloroform 80ml

[0036] 4. Isopropanol 80ml

[0037] 5. DEPC water 10ml

[0038] 6. Mixture of 10×random primers and Oligo dT primers 100μl

[0039] 7. 5× Reverse Transcription Reaction Buffer 150μl

[0040] 8. 10mM with Mg 2+100 μl of deoxyribonucleotide triphosphate dNTP

[0041] 9. 200U / μl M-MLV reverse transcriptase 50μl

[0042] 10. SYBR Green qPCR Mix 500μl

[0043] 11. 3μM target gene Lnc-DCTD-25 specific primer (its sequence is shown in SEQ NO: 2, 3) 50μl

[0044] 12. 3μM internal reference gene TUBA1A-specific primer (its sequence is shown in SEQ NO: 4, 5) 50μl

Embodiment 2

[0045] Example 2 Detection of Lnc-DCTD-25 in Tissue Samples

[0046] 1. Collect the pancreatic 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.

[0047] 2. Extraction of RNA in tissues:

[0048] (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 thoroughly mixed evenly.

[0049] (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 12,000 rpm for 10 minutes.

[0050] (3) Take the upper aqueous phas...

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Abstract

The invention relates to pancreatic cancer prognosis molecular marker non-coding RNA Lnc-DCTD-25 and application thereof, which can effectively solve the problem for preparing a kit for predicting theprognosis of a pancreatic cancer patient. A transcript length of the pancreatic cancer prognosis molecular marker non-coding RNA Lnc-DCTD-25 is greater than 200 nucleotides, a sequence is SEQ NO.1. The pancreatic cancer prognosis molecular marker non-coding RNA Lnc-DCTD-25 is used for preparing a pancreatic cancer prognosis real-time fluorescent quantitative PCR detection kit. The kit comprises akit expression specific primer arranged in a box body and used for detecting the prognosis of the pancreatic cancer patient, a primer expressed by an inner reference gene TUBA1A, RNA extracted from pancreatic cancer tissues and a reagent for performing the reverse transcription and fluorescent quantitative PCR, wherein the sequence of the specific primer is SEQ NO. 2 and 3, and the sequence of the primer expressed by the inner reference gene TUBA1A is No. 4 and 5. The non-coding RNA Lnc-DCTD-25 is used as a novel molecular marker of the pancreatic cancer prognosis, can predict the prognosis of the pancreatic cancer in time, can treat the pancreatic cancer patient in time, can prolong the living time of the patient, can improve the survival rate, and is a great innovation for the prognosisof the pancreatic cancer.

Description

technical field [0001] The invention relates to tumor molecular biology, in particular to a pancreatic cancer prognosis molecular marker non-coding RNA Lnc-DCTD-25 and its application. Background technique [0002] Pancreatic cancer is a highly malignant tumor with increasing incidence, poor prognosis and low 5-year survival rate. Pancreatic cancer is the fourth leading cause of cancer death in the United States. In my country, pancreatic cancer ranks sixth in cancer mortality. The reasons are mainly related to the low rate of early diagnosis, invasiveness, and insensitivity to radiotherapy and chemotherapy. Although the existing diagnostic methods are mainly imaging examinations such as B-ultrasound, endoscopic ultrasonography, abdominal magnetic resonance and cholangiopancreatography, and abdominal computed tomography, etc., when the index rises or symptoms appear, imaging examinations are performed when the condition has reached the middle stage. At an advanced stage, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/113
CPCC12Q1/6886C12Q2600/118C12Q2600/158C12Q2600/166C12Q2600/178
Inventor 田田
Owner THE FIRST AFFILIATED HOSPITAL OF ZHENGZHOU UNIV