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Molecular marker for diagnosis and treatment of pituitary adenoma

A pituitary tumor and drug technology, applied in the field of biomedicine, can solve the problems of low pituitary function, unclear mechanism and mechanism, affecting the normal secretion function of the pituitary gland, etc.

Active Publication Date: 2016-07-20
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Surgical treatment also includes transnasal sphenoidal endoscopic lesion resection and craniotomy to explore sellar region tumor resection, but various treatment options currently used can affect the normal secretory function of the pituitary gland, and even cause hypopituitarism. Hormone replacement therapy, painful
With the development of biomedicine and molecular biology, the research on the pathogenesis of pituitary tumors has made great progress recently, but the exact mechanism and mechanism are still unclear, so further extensive research on the mechanism of occurrence and development of pituitary tumors will continue become a research hotspot

Method used

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  • Molecular marker for diagnosis and treatment of pituitary adenoma
  • Molecular marker for diagnosis and treatment of pituitary adenoma
  • Molecular marker for diagnosis and treatment of pituitary adenoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Example 1 Screening for gene markers associated with pituitary tumors

[0063] 1. Sample collection

[0064] Six samples of normal pituitary tissue and pituitary tumor tissue were collected. All the above specimens were obtained with the consent of the organizational ethics committee.

[0065] 2. RNA sample preparation (using miRNAkit for operation)

[0066] The tissues obtained above were shredded, put into liquid nitrogen and ground into powder, and RNA was extracted and isolated according to the instructions in the kit. details as follows:

[0067] 1) Isolation of RNA:

[0068] A. Add in tissue homogenate or cells Reagent II 1ml;

[0069] B. Stand at room temperature for 3 minutes, add 0.2ml chloroform and shake vigorously for 15 seconds;

[0070] C. Place on ice for 10 minutes;

[0071] D. Centrifuge at 12000g for 15min at 4°C;

[0072] E. Transfer 80% of the aqueous phase into a new 2ml EP tube, add 1 / 2 the amount of absolute ethanol, and shake;

[007...

Embodiment 2

[0087] Example 2 QPCR sequencing to verify the differential expression of the PAEP gene

[0088] 1. Large-sample QPCR verification of PAEP gene differential expression. According to the sample collection method in Example 1, 50 cases of normal pituitary tissues and 50 cases of pituitary tumor tissues were selected.

[0089] 2. The RNA extraction steps are as described in Example 1.

[0090] 3. Reverse transcription:

[0091] 1) Reaction system:

[0092] Reagent

volume

MgCl 2

2μl

10×RT Buffer

1μl

RNase-free water

3.75μl

dNTP mix

1μl

RNase inhibitor

0.25μl

AMV reverse transcriptase

0.5μl

Oligo-dT Adapter Primer

0.5μl

Experimental sample

1μl

[0093] 2) Reverse transcription reaction conditions

[0094] According to the reverse transcription reaction conditions in RNAPCRKit (AMV) Ver.3.0.

[0095] 42°C~55°C for 60 minutes, 99°C for 2 minutes, 5°C for 5 minutes.

[0...

Embodiment 3

[0113] Overexpression of embodiment 3PAEP gene

[0114] 1. Cell culture

[0115] Human pituitary tumor cell line GT1.1 was incubated at 37°C and 5% CO in medium 1640 containing 10% fetal bovine serum and 1% P / S. 2 , Cultivated in an incubator with a relative humidity of 90%. Change the medium once every 2-3 days, and use 0.25% EDTA-containing trypsin for routine digestion and passage.

[0116] 2. siRNA design

[0117] siRNA sequence against PAEP gene:

[0118] Negative control siRNA sequence (siRNA-NC):

[0119] The sense strand is 5'-UUCUCCGAACGUGUCACGU-3' (SEQ ID NO.7),

[0120] The antisense strand is 5'-ACGUGACACGUUCGGAGAA-3' (SEQ ID NO.8);

[0121] siRNA1-PLD5:

[0122] The sense strand is 5'-AACUUCUUUGGAUUCUCAGUC-3' (SEQ ID NO.9),

[0123] The antisense strand is 5'-CUGAGAAUCCAAAAGAAGUUCA-3' (SEQ ID NO.10);

[0124] siRNA2-PLD5:

[0125] The sense strand is 5'-UUGAUCUUGAACUUCUUUGGA-3' (SEQ ID NO.11),

[0126] The antisense strand is 5'-CAAAGAAGUUCAAGAUCAACU-3'...

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Abstract

The invention discloses a molecular marker for diagnosis and treatment of pituitary adenoma. Pituitary adenoma is a common intracranial benign tumor and can induce endocrine symptoms as well as intracranial hypertension and change of vision and view. Experiments prove that the expression of PAEP gene in the pituitary adenoma tissue is up-regulated, and the expression of the PAEP gene can inhibit the proliferation of pituitary adenoma cells and promote the apoptosis of the pituitary adenoma cells. The invention provides a new molecular means for the diagnosis and treatment of pituitary adenoma.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a molecular marker for diagnosing and treating pituitary tumors. The molecular marker is PAEP gene. Background technique [0002] Pituitary tumor is one of the common neuroendocrine tumors that can lead to the imbalance of pituitary hormone secretion, accounting for about 10%-15% of central nervous system tumors. The vast majority of pituitary tumors are benign, and pituitary tumors are common intracranial tumors that can cause significant morbidity through mass effect and / or inappropriate secretion of pituitary hormones. [0003] According to the characteristics of tumor cell staining, pituitary tumors can be divided into oncocytic adenomas, basophilic adenomas and chromophobe adenomas. According to the classification of endocrinology, pituitary tumors can be divided into prolactinoma, growth hormone adenoma, corticotropin adenoma, gonadotropin adenoma, thyroid-stimulating hormone adeno...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/68G01N33/574A61K45/00A61K48/00A61K31/713A61P35/00
CPCA61K31/713A61K45/00C12Q1/6886C12Q2600/158G01N33/57407G01N33/68
Inventor 杨承刚王欣月任静
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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