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Application of NFIC gene in preparation of diagnostic and therapeutic products for pituitary adenomas

A technology for pituitary tumors and products, applied in the field of NFIC gene diagnosis and treatment of pituitary tumors, can solve the problems of high postoperative recurrence rate, difficult complete resection, and high recurrence rate

Active Publication Date: 2016-08-10
BEIJING NEUROSURGICAL INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For most pituitary tumors, transsphenoidal microsurgery is still the preferred treatment method. Although the complications of transsphenoidal microsurgery are reduced compared with traditional craniotomy, the operation cannot completely relieve the patient's problems. The recurrence rate is high, and the recurrence rate of large adenomas (diameter>lcm) is as high as 5 years after surgery
Invasive pituitary adenomas are difficult to completely remove by surgery, and the recurrence rate is higher, which is a major problem in clinical treatment

Method used

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  • Application of NFIC gene in preparation of diagnostic and therapeutic products for pituitary adenomas
  • Application of NFIC gene in preparation of diagnostic and therapeutic products for pituitary adenomas
  • Application of NFIC gene in preparation of diagnostic and therapeutic products for pituitary adenomas

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Example 1 Differential expression of NFIC gene

[0060] 1. Collect samples

[0061] Patients with pituitary tumors: Among the 10 patients with pituitary tumors, there were 5 males, aged 28-73, with an average age of 47; 5 females, aged 25-68, with an average age of 44. Inclusion criteria: patients underwent transsphenoidal pituitary tumor resection, brain MRI and other imaging data were available before surgery, and postoperative pathological sections confirmed pituitary tumor. With the consent of the hospital ethics committee, the materials were collected after the patients signed the informed consent.

[0062] Normal pituitary gland tissue: 5 cases were taken from the Anatomy Department of the University, and endocrine-related diseases were excluded. The sampling method was the same as that of patients with pituitary tumors.

[0063] The required tissues were taken during the operation, washed with normal saline, immediately put into cryopreservation tubes treated ...

Embodiment 2

[0074] Example 2 qPCR experiment to verify the differential expression of NFIC gene

[0075] 1. Collect samples

[0076] According to the standard of Example 1, 40 patients with pituitary tumors were collected, and their pituitary tumor tissues were collected. 30 normal pituitary tissues were collected.

[0077] 2. RNA extraction

[0078] use Reagent (invitrogen, Cat. No. 15596-018) was used for sample RNA extraction. The specific operation is as follows:

[0079] Collect the samples and freeze them in liquid nitrogen. After taking them out, put the tissue into a pre-cooled mortar for grinding. After the tissue sample is powdered:

[0080] ① Add Trizol and store at room temperature for 5 minutes;

[0081] ② Add 0.2ml of chloroform, vibrate the centrifuge tube vigorously, mix well, and place it at room temperature for 5-10 minutes;

[0082] ③ After 15 minutes of high-speed centrifugation at 12000rpm, suck the upper aqueous phase (absorb 70%) into another new centrifuge ...

Embodiment 3

[0101] Embodiment 3 inhibits NFIC gene expression

[0102] 1. siRNA design and synthesis

[0103] siRNA sequences targeting NFIC:

[0104] siRNA1-NFIC:

[0105] The sense strand is 5'-UCUUCUUCUUCUUCUUCUGCC-3' (SEQ ID NO.7);

[0106] The antisense strand is 5'-CAGAAGAAGAAGAAGAAGAAA-3' (SEQ ID NO.8),

[0107] siRNA2-NFIC:

[0108] The sense strand is 5'-AGAUGAAUAACUUAGGAAGAC-3' (SEQ ID NO.9);

[0109] The antisense strand is 5'-CUUCCUAAGUUAUUCAUCUCC-3' (SEQ ID NO.10),

[0110] siRNA3-NFIC:

[0111] The sense strand is 5'-UAUUGUUUAAAGGUAAUACUU-3' (SEQ ID NO.11);

[0112] The antisense strand is 5'-GUAUUACCUUUAAACAAUAUC-3' (SEQ ID NO.12)

[0113] The above siRNA sequences and negative control siRNA sequences (siRNA-NC) were provided by Shanghai Gemma Pharmaceutical Technology Co., Ltd.:

[0114] 2. Culture and transfection of pituitary tumor cells

[0115] 2.1 Cell culture

[0116] The pituitary tumor tissue pieces were immersed in RPMI-1640 medium, washed 3 times with ...

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Abstract

The invention discloses NFIC gene capable of serving as a molecular marker for early diagnosis of pituitary adenomas. Experiments of the invention prove that NFIC gene has significantly improved expression in pituitary adenomas tissues when compared to normal pituitary tissues; RNA interference experiments prove that NFIC can affect the proliferation of pituitary adenomas cells. According to the study results of the invention, it is possible to develop drugs capable of inhibiting NFIC gene expression, thus preventing and treating pituitary adenomas clinically.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of NFIC gene in the diagnosis and treatment of pituitary tumors. Background technique [0002] Pituitary adenoma is a common intracranial tumor, ranking third in the incidence of intracranial tumors, second only to glioma and meningioma, accounting for about 10-25% of intracranial tumors, although the pituitary adenoma Tumor is a benign tumor, but some tumors grow aggressively and often involve the optic nerve, sphenoid sinus, cavernous sinus, and internal carotid arteryC 2 Segment, hypothalamus and other surrounding important structures. Affect the normal endocrine and neurological functions of patients, such as amenorrhea, lactation, infertility, sexual dysfunction, acromegaly, hypercortisolism, decreased vision, visual field defects, etc. For most pituitary tumors, transsphenoidal microsurgery is still the preferred treatment method. Although the complications of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/68G01N33/574A61K45/00A61K39/395A61K31/7088A61P35/00
CPCA61K31/7088A61K39/3955A61K45/00C12Q1/6886C12Q2600/158G01N33/57407G01N33/68
Inventor 赵澎张亚卓李储忠白吉伟桂松柏王红云李丹何乐刘潜
Owner BEIJING NEUROSURGICAL INST
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