CN109528749B

CN109528749B - Application of long-chain non-coding RNA-H19 in preparation of drug for treating pituitary tumor

Info

Publication number: CN109528749B
Application number: CN201710877845.2A
Authority: CN
Inventors: 吴哲褒; 苏冰; 吴泽睿; 颜立冲; 刘衍挺; 赵卫国
Original assignee: Ruinjin Hospital Affiliated to Shanghai Jiaotong University School of Medicine Co Ltd
Current assignee: Ruinjin Hospital Affiliated to Shanghai Jiaotong University School of Medicine Co Ltd
Priority date: 2017-09-22
Filing date: 2017-09-22
Publication date: 2023-04-28
Anticipated expiration: 2037-09-22
Also published as: CN109528749A

Abstract

The invention belongs to the technical field of biological medicines, relates to a new application of long-chain non-coding RNA-H19 in preparing a medicament for treating pituitary tumor, and in particular relates to an effect of inhibiting occurrence and development of pituitary tumor in vivo and in vitro of long-chain non-coding RNA-H19 and deep mechanism research thereof, thereby achieving the purpose of treating the pituitary tumor. The results of in vitro cell culture, virus-mediated gene regulation experiments and rat in-situ pituitary tumor H19 injection model experiments show that H19 inhibits the growth of tumor cells by inhibiting the phosphorylation of 4E-BP1 at the downstream of an mTOR signal path, thereby achieving the purpose of inhibiting the growth of tumor cells. The long-chain non-coding H19 can be used for preparing medicines for treating pituitary tumors, is clinically used for treating the pituitary tumors, and particularly can remarkably inhibit tumor growth to achieve a therapeutic effect. The invention also provides a new strategy reference for clinically treating pituitary tumors, especially refractory pituitary tumors.

Description

Application of long-chain non-coding RNA-H19 in preparation of drug for treating pituitary tumor

Technical Field

The invention belongs to the technical field of biological medicines, and relates to a small molecular medicinal application of long-chain non-coding RNA (lncRNA) -H19. In particular to the application of H19 in preparing a drug for treating pituitary tumor.

Background

According to medical statistics investigation, the annual incidence rate of the population suffering from pituitary adenomas in China is as high as 7.5-15/10 ten thousand, wherein the pituitary adenomas discovery rate is 10% -38.5% (average 22.5%) when the normal population is examined by a stochastic Magnetic Resonance Imaging (MRI) technology; according to 14 hundred million demographics of the population in China, more than 15 to 30 ten thousand new pituitary adenomas are present each year, wherein the prolactinadenomas are the most common subtype in the pituitary adenomas and account for about 50% of all pituitary adenomas. Studies have shown that with advances in medical technology, the level of diagnosis is increasingly advanced, microsurgery and endoscopic techniques are evolving, and the statistics of the incidence of pituitary tumors are still increasing. Pituitary adenomas cause disorders of endocrine function such as amenorrhea-lactation and infertility, giant person and acromegaly, cushing syndrome, etc.; and the clinical symptoms seriously jeopardize the survival and life quality of patients due to the fact that the pituitary tumors press adjacent structures to increase, the vision is reduced, the pituitary functions are low and the like, and on the other hand, the great economic pressure and the medical resource pressure are brought to the country and the families of the patients due to the huge number of patients.

In clinical practice, the prolactinoma is the first choice of drug treatment, wherein dopamine receptor agonists (Dopaminergic Agonist, DA) are the first choice of clinical treatment, bromocriptine is commonly used in China, and cabergoline is commonly used in European and American countries. The medicine can effectively reduce tumor volume and restore normal Prolactin (PRL) level of 80% -90% of patients, but about 10% -20% of patients are insensitive to bromocriptine and even cabergoline treatment, and the part of cases are called drug-resistant cases in the industry, and clinical practice shows that even though the drug-resistant patients are treated by surgery, the patients still face the problems that the hyper-prolactin can not restore normal after surgery, tumor recurrence and the like. In addition, other types of pituitary adenomas, such as growth hormone adenomas, ACTH adenomas, TSH adenomas, and the like, lack effective pharmacological treatment, and a significant proportion of patients are refractory pituitary adenomas; thus, resistant prolactinoma and refractory pituitary tumors remain a great clinical challenge in the art.

Long non-coding RNA (lncRNA) is a family of single-stranded RNAs greater than 200 nucleic acid sequences in length, and has the effect of regulating gene expression. Research shows that lncRNA participates in the processes of apoptosis regulation, tumor infiltration, metastasis and the like; in addition, the growth of tumor cells is also affected by means of epigenetic regulation, thus playing an important physiological role in tumorigenesis and development and treatment. In recent years, there have been studies on the continued discovery of some abnormal expressed lncRNA in tumor tissues, involving tumors of various systems, of which lncRNA-H19 is one of the important lncRNA found earliest and involved in the development of tumorigenesis. H19 is the product of the imprinted gene of insulin-like growth factor2 (insulin-like growth factor, igf 2), expressed only in embryonic tissues, not in adult tissues; it has been reported that some tumors are associated with the lack of expression of H19, such as Wilms' tumor, myeloma, adrenal tumor, etc. Meanwhile, the inventor of the application discovers and verifies in a tissue sample through a gene chip technology in the early stage that H19 is significantly low-expressed in pituitary tumor, which indicates that H19 may have the effect of inhibiting the growth of the pituitary tumor.

Mammalian target rapamycin (mammalian target of rapamycin, mTOR) is a threonine/serine kinase that plays an important role in cell growth, differentiation, and regulation of cell cycle, among other aspects. mTOR signaling is closely related to tumorigenesis, such as prostate cancer, lung cancer, gastric cancer, and intracranial tumors such as gliomas and pituitary tumors. mTOR inhibitors are capable of inhibiting tumor growth and angiogenesis due to abnormalities in this signaling pathway, and inhibition of mTOR has been the basis for clinical drug therapy for malignant tumors. Thus, the penetration of mTOR research is of great importance for tumor-targeted therapies. Activation mTOR is involved in multiple cellular functions by phosphorylating certain factors during protein translation, mainly 4E-BP1 (eIF 4E-binding protein 1) and S6K1 (ribosome protein subunit kinase 1). The 4E-BP1 achieves inhibition of translation initiation by competitively binding eIF-4G to eIF-4E, whereas phosphorylated 4E-BP1 accelerates dissociation of 4E-BP1 from the eIF-4G and eIF-4E complex, thereby accelerating the translation process. However, there are few studies concerning the lncRNA and mTOR signaling pathway, and although lncRNA-00961 has been reported to be able to effectively inhibit the mTOR pathway, the regulatory relationship between H19 and mTOR has not been reported.

Based on the current state of the art, the inventor of the application intends to provide a new medicinal application of long-chain non-coding RNA-H19, in particular to a new application of H19 in preparing a drug for treating pituitary tumor.

Disclosure of Invention

The invention aims to provide a novel medicinal application of long-chain non-coding RNA-H19 based on the current state of the art, and in particular relates to a novel application of H19 in preparation of a drug for treating pituitary tumor.

In the invention, the long non-coding RNA-H19 has a length of 2369 nucleotides in a rat and a length of 2362 nucleotides in a human, and the similarity of the two is 73%.

The long-chain non-coding RNA-H19 can effectively inhibit the growth of pituitary tumors in vivo and in vitro, and experiments prove that the long-chain non-coding RNA-H19 can inhibit the proliferation of tumor cells by inhibiting mTOR signaling pathway, thereby achieving the purpose of treating the pituitary tumors.

In the embodiment of the invention, in vitro experiments are carried out by adopting lentivirus to overexpress H19 genes in a pituitary tumor cell line GH3 cell, and the results show that the gene has an inhibition effect on cell growth; further, the transplanting tumor model and the in-situ tumor forming model prove that the H19 has the effect of inhibiting the growth of the GH3 cell in the subcutaneous transplanting tumor model and the rat lactalbumin tumor in-situ growth model; meanwhile, the subcutaneous transplantation tumor model experiment shows that the inhibition effect of H19 on GH3 tumor formation in vivo is better than that of cabergoline; the mechanism research results show that: h19 is capable of inhibiting pituitary tumor and pituitary tumor cell growth by inhibiting the phosphorylation level of the downstream substrate 4E-BP1 of mTORC1 in the cells, thereby inhibiting mTORC1 function;

in vitro, through cell culture, virus-mediated gene regulation experiments and rat in-situ pituitary tumor H19 injection model experiments, the results show that H19 inhibits the growth of tumor cells by inhibiting the phosphorylation of 4E-BP1 at the downstream of an mTOR signal path, thereby achieving the purpose of inhibiting the growth of the tumor cells;

the experimental result shows that the long-chain non-coding H19 can be used for preparing medicines for treating pituitary tumors, is clinically used for treating the pituitary tumors, and particularly can remarkably inhibit tumor growth to achieve the therapeutic effect. The invention also provides a new strategy reference for clinically treating pituitary tumors, especially refractory pituitary tumors.

The present invention will be described in detail below with reference to specific drawings and examples for the purpose of facilitating understanding. It is specifically pointed out that the specific examples and the figures are for illustrative purposes only and that it is obvious that within the scope of the invention various modifications and changes can be made by a person skilled in the art based on the description herein, which modifications and changes are also within the scope of the invention.

Drawings

Fig. 1 shows: h19 can inhibit pituitary tumor GH3 cells, and meanwhile, a nude mouse transplantation tumor model proves that H19 can effectively inhibit subcutaneous tumor formation of GH3 cells.

Fig. 2 shows: and constructing a rat in-situ pituitary tumor model by utilizing an estrogen induction mode, and injecting the H19 over-expression adenovirus into the rat pituitary tumor by utilizing a targeting injection technology, so that the H19 can effectively inhibit the growth of the rat in-situ pituitary tumor.

Fig. 3 shows: through protein level experiments, the H19 can effectively inhibit the phosphorylation level of 4E-BP1 so as to inhibit mTorrC 1 activity, and it is clear that the H19 regulates cell activity and GH3 cell neoplasia through the phosphorylation of 4E-BP 1.

Fig. 4 shows: by using a subcutaneous nodulation experiment of nude mice, compared with cabergoline, the inhibition effect of H19 on GH3 nodulation is obviously enhanced, and meanwhile, the inhibition effect of H19 on 4E-BP1 phosphorylation level is also stronger than that of cabergoline.

Detailed Description

Example 1, H19 experiments for inhibiting growth of pituitary tumor cell lines in vitro and in vivo

Experimental materials:

rat pituitary tumor cell line GH3 (purchased from ATCC) and human primary pituitary adenoma cells (after surgical removal and direct culture of the culture broth) were cultured at 37℃with 5% CO ₂ Conventional culture under conditions, the medium was DMEM/F12 (Gibco) containing 2.5% fetal bovine serum (Gibco) plus 12.5% horse serum. Lentiviruses over-expressing the H19 gene were constructed in Shanghai Ji Kai Biotechnology Inc. MTS assay cell Activity reagents were purchased from Promega (USA). The crystal violet staining solution was purchased from a biological source (china). Nude mice were purchased from SLAC (Shanghai).

The experimental method comprises the following steps:

1) MTS assay to examine the effect of H19 on GH3 cell proliferation potency

(1) H19 overexpressed GH3 cells and normal control GH3 cells were pre-suspended in conventional medium to adjust cell density to 0.50x10 ⁴ Cells/well were seeded in 96 well plates for 12 hours in an adherent treatment with 5 sub-wells per group, 100 μl of system per well, and 10 μl of MTS assay reagent was added;

(2) Cells were incubated at 37℃with 5% CO ₂ Culturing for 1-4 hours under the condition; determining the initial living cell number by measuring the value of absorbance at a detection wavelength of 490 nm;

(3) The cells were returned to 37℃and 5% CO ₂ Culturing for 24 hours, 48 hours, 72 hours and 92 hours under the condition;

(4) 10 microliters of MTS assay reagent was added at each time point, the number of viable cells was calculated according to the method (2), the comparison of the two sets of cell activities was determined, N=5, and the experiment was performed three times in parallel, and the results of GH3 cell activities are shown in FIG. 1. A.

2) Plate cloning experiments to examine the effect of H19 on the growth of GH3 cell clones

(1) Suspending GH3 cells over-expressed by H19 and common control GH3 cells in a conventional culture medium, adjusting the cell density to 500 cells/hole, inoculating the cells to a 6-hole culture plate, and transfecting a GH3 cell strain of a blank lentivirus by a blank control group;

(2) Culturing the above cells at 37deg.C, 5% CO ₂ Under the condition, the culture time is 2-3 weeks;

(3) Observing the clone formation condition every 2 days during the period, and stopping culturing when the clone formation is visible to naked eyes;

(4) Dyeing with crystal violet staining solution for 15 minutes at room temperature, cleaning the staining solution with clear water, taking a picture to leave a result, counting the number of cloning points by picture software, and performing experiments for three times; the cell clone formation is shown in FIGS. 1.B, C.

3) Nude mouse transplanted tumor model experiment to determine in vivo H19 effect on GH3 cell neoplasia

(1) Selecting a nude mouse with the age of 4 weeks;

(2) Collecting logarithmic growth of H19-over-expressing GH3 cells and normal GH3 cells, centrifuging at 1000 rpm with 10ml of serum-free medium for 3min, washing for 3 times, and resuspending tumor cells with serum-free medium to a density of 1×10 ⁷ Cells/ml, 100ul (i.e., 1X 10) of cells were inoculated in each nude mouse in the armpit ⁶ Tumor cells), palpable in the armpit of nude mice for about 4 days;

(3) The sizes (unit: mm) of the long axis and the wide axis of the tumor are measured by a vernier caliper every day;

(4) After raising for 14-21 days, the nude mice are treated conventionally, tumor body is obtained, weighed and photographed, tumor weight and tumor volume are counted, and tumor volume = long axis x wide axis ² X 1/2; in vivo, the experimental results of GH3 cells are shown in FIGS. 1.D and E, F;

the experimental results show that H19 can inhibit the growth of the pituitary tumor GH3 cells in vitro and in vivo.

Example 2 in situ Pituitary tumor H19 injection model experiment in rats

Experimental materials:

fischer344 rats were purchased from Beijing Veityle Liwa laboratory animal Co.Ltd and bred in Beijing Tiantan Hospital neurosurgery institute; 17-beta estradiol is available from sigma corporation (U.S.); adenovirus over-expressing H19 gene is constructed in Shanghai Heng biological company;

the experimental method comprises the following steps:

h19 in-situ pituitary tumor treatment experiment on rats

(1) Selecting 4-week-old female Fischer344 rats, and dividing 10 rats into an H19 treatment group and a control group, wherein each group comprises 4 rats;

(2) Conventional subcutaneous implantation of 17-beta estradiol tablets containing 10mg per mouse, and culture for 6 weeks induced rat pituitary tumor model; at week 6, a Magnetic Resonance Imaging (MRI) is adopted to confirm that an in-situ model of the rat pituitary tumor is successfully constructed;

(3) Anaesthetizing rats with 10% chloral hydrate (3.5 ml/kg), and injecting adenovirus with H19 over-expressed gene or empty plasmid into tumor under the guidance of microscopic targeting injection technique by using 10UI adenovirus (10-12 order of magnitude);

(4) Continuously culturing for 4-6 weeks, measuring tumor size by magnetic resonance imaging, and taking out a sample after anesthesia;

the experimental results showed that H19 was able to significantly reduce the size of the rat in situ pituitary tumor (experimental results are shown in fig. 2.A, and statistical data graphs are shown in fig. 2. B).

Example 3H 19 influences on cellular Activity by inhibiting 4E-BP1 phosphorylation

Experimental materials:

RIPA lysate, PMSF, loading buffer (5×), BCA protein quantification cassette, DAPI purchased from the b.clouds biotechnology institute (Jiangsu); protease Inhibitor Cocktail from Merck company (Germany). BSA was purchased from sigma (9048-46-8); PBS, 0.5% Trypsin-EDTA, available from Gibco;

antibody: tubulin (10068-1-AP, proteintech), p-4EBP1 Thr70 (9455S, CST), 4E-BP1 (9644S, CST), p-4EBP1 Thr37/46 (9459S, CST), S6K1 (9202S, CST), p-S6K1 (9205S, CST), AKT (9272, CST), p-AKT (9272, CST), and the high intensity chemiluminescent kit was purchased from Bio-rad.

The experimental method comprises the following steps:

1) Cell protein extraction and detection of each target protein

(1) GH3 cells were suspended in conventional medium and cell density was adjusted to 4X 10 ⁵ The cells/well are inoculated in a 6-well culture plate for adherence overnight, each well contains 2ml of culture medium, the final concentration of the drug is 25 mu M, and the cells/well respectively act for 0h, 12h, 24h, 48h and 0h as a control group, contain 0.3% of DMSO and contain 5% CO at 37 DEG C ₂ Culturing under the condition;

(2) WesternBlot detects the expression of p-AKT, p-S6K1, p-4EBP 1. PMSF (100 mM, 1:100) and Proteaseln inhibitor CocktailSetIII (1:200) were added to RIPA lysate to collect cells at the time point of drug action, and when cells were collected, the culture solution in 6-well plates was removed, after gently washing the cells with PBS, GH3 cells were digested with 0.05% pancreatin for 10 seconds, pancreatin was removed, 1ml complete medium was added to neutralize, collected in 1.5ml EP tube, centrifuged at 2000rpm for 5min, the supernatant was discarded, and 100. Mu.l lysate was added for 30min of ice bath lysis; centrifuging at 12000rpm for 20min after cracking; taking out the supernatant for quantification, simultaneously taking out part of the supernatant, adding 1/4 volume of loading buffer solution (5×), and boiling at 100 ℃ for 10min; storing the denatured protein in a refrigerator at-80 ℃;

(3) Sample loading amount: the BCA protein quantitative box is used for quantifying each group of extracted protein samples, and the loading amount of the corrected protein samples is 50 mug;

(4) Electrophoresis: carrying out 80V constant-pressure electrophoresis for 30 minutes to concentrate a sample; separating samples after electrophoresis at 120V for 1.5 hours, and judging the electrophoresis degree according to a marker;

(5) Transferring: wet turning, 170mA, constant-current film turning for 2 hours;

(6) Antibody hybridization: first blocked with 5% BSA for 1 hour on a room temperature shaker; diluting primary antibody (Tubulin 1:5000; p-AKT 1:2000; p-S6K 11:1000; p-4EBP 1:1000; etc.) with blocking solution, and hybridizing the transferred membrane in a hybridization tank at 4deg.C overnight; TBST washing the membrane 3 times (shaking table at room temperature, 2mL each time, 5 min); diluting the secondary antibody by using a sealing solution, and carrying out shaking table hybridization for 1 hour at room temperature in a hybridization groove; performing chemiluminescence detection after washing the TBST film for three times;

(7) Chemiluminescent detection: detecting target proteins by adopting an enhanced chemiluminescence method (ECL method);

the experimental results are shown in fig. 3.A, 3.G.

2) Extraction and detection of tissue protein extraction protein

(1) After anesthesia, mice are sacrificed, tumors are stripped rapidly by using an dissecting instrument, a small amount of specimens are taken and put into a 1.5ml clean EP tube, and liquid nitrogen is put rapidly;

(2) Taking out the tissue specimens in the liquid nitrogen after all the specimens are taken, and adding a proper amount of RIPA lysate;

(3) Fully grinding the tissue on ice of the tissue homogenizer, and centrifuging at 12000rpm for 20min after the tissue homogenizer is cracked; taking out the supernatant for quantification, simultaneously taking out part of the supernatant, adding 1/4 volume of loading buffer solution (5×), and boiling at 100 ℃ for 10min; storing the denatured protein in a refrigerator at-80 ℃;

(5) The results of the experiments for detecting the expression level of the tissue protein and the precautions are shown in FIG. 3. B.

3) Nude mice transplanted tumor model experiment determines that H19 inhibits development of pituitary tumor through 4E-BP1 in vivo

(1) Selecting a nude mouse with the age of 4 weeks;

(2) Collecting logarithmic growth normal GH3 cells, H19-overexpressing GH3 cells and GH3 simultaneously overexpressing and intervening 4E-BP1, centrifuging for 3min at 1000 rpm with 10ml of serum-free medium, washing for 3 times continuously, and resuspending tumor cells with serum-free medium to a density of 1×10 ⁷ Cells/ml, 100ul (i.e., 1X 10) of cells were inoculated in each nude mouse in the armpit ⁶ Tumor cells), palpable in the armpit of nude mice for about 4 days;

(3) The sizes (unit: mm) of the long axis and the wide axis of 3 groups of tumors are measured by a vernier caliper every day;

(4) After raising for 14-21 days, the nude mice are treated conventionally, tumor body is obtained, weighed and photographed, tumor weight and tumor volume are counted, and tumor volume = long axis x wide axis ² X 1/2; the experimental results are shown in fig. 3.D, e, f;

experimental results show that H19 inhibits pituitary tumor through 4E-BP1 pair.

4) Immunohistochemical detection of the Effect of H19 on phosphorylation of rat Pituitary tumor 4E-BP1 in situ

(1) Dewaxing paraffin sections to water;

(2)3％H ₂ O ₂ incubating for 5-10 minutes at room temperature to eliminate the activity of endogenous peroxidase;

(3) Washing with distilled water, and soaking in PBS for 5min x2 (if antigen retrieval is needed, this step can be followed);

(4) 5% BS was blocked, incubated at room temperature for 10min, serum was decanted, and no washing was performed. Dripping primary antibody working solution, and incubating at 37 ℃ for 1-2 hours or overnight at 4 ℃;

(5) PBS rinse, 5min x3 times. A proper amount of biotin-labeled secondary antibody working solution is dripped, incubated for 10-30 minutes at 37 ℃, washed by PBS (phosphate buffer solution) for 5 minutes x3 times;

(6) Dripping a proper amount of horseradish enzyme or alkaline phosphatase labeled streptavidin working solution, incubating for 10-30 minutes at 37 ℃, flushing with PBS (phosphate buffer solution), and carrying out 5-3 times for a minute;

(7) The DAB color-developing agent is developed for 10 minutes (DAB or NBT/BCIP);

(8) Washing with tap water, counterstaining, dewatering, transparency, sealing, and taking pictures; the experimental results are shown in fig. 3. C;

the experimental result shows that H19 can inhibit 4E-BP1 phosphorylation in vivo.

Example 4 in vivo verification of H19 inhibition of pituitary tumor in nude mice over cabergoline

Experimental materials:

nude mice purchase channels as described above, cabergoline purchased from Tocres corporation (USA).

The test method comprises the following steps:

h19 inhibits the subcutaneous GH3 tumor formation effect of nude mice better than cabergoline

(1) Selecting a nude mouse with the age of 4 weeks;

(2) Collecting logarithmic growth normal GH3 cells and H19-overexpressing GH3 cells, centrifuging at 1000 rpm with 10ml of serum-free medium for 3min, washing for 3 times, and resuspending tumor cells with serum-free medium to a density of 1×10 ⁷ Cells/ml, 100ul (i.e., 1X 10) of cells were inoculated in each nude mouse in the armpit ⁶ Tumor cells), about 4 days atArmpit palpation and tumor of nude mice; wherein normal GH3 cells are divided into two groups, including a control group and a cabergoline-treated group;

(3) The cabergoline treatment group is subjected to intragastric cabergoline treatment every other day, and the dosage concentration is 0.75mg/kg;

(4) The sizes (unit: mm) of the long axis and the wide axis of 3 groups of tumors are measured by a vernier caliper every day;

(5) After raising for 14-21 days, the nude mice are treated conventionally, tumor body is obtained, weighed and photographed, tumor weight and tumor volume are counted, and tumor volume = long axis x wide axis ² X 1/2; the experimental results are shown in fig. 4.A, b, c.

2) H19 has better effect of inhibiting 4E-BP1 phosphorylation than cabergoline in nude mice

The method for extracting the tissue protein is similar to that described above, and the phosphorylation level of p-4E-BP1 in a normal group, an H19 over-expression group, a cabergoline treatment group and a nude mouse transplanted tumor is detected respectively, and the experimental result is shown in a graph 4.D.

The results of in vitro cell culture, virus-mediated gene regulation experiments and rat in-situ pituitary tumor H19 injection model experiments show that H19 inhibits the growth of tumor cells by inhibiting the phosphorylation of 4E-BP1 at the downstream of an mTOR signal path, thereby achieving the purpose of inhibiting the growth of tumor cells.

Claims

1. Use of long-chain non-coding RNA-H19 in preparing a medicament for treating pituitary tumor;

the long-chain non-coding H19 has a nucleotide length of 2369 in rats and 2362 in humans

A nucleotide, both of which are 73% similar;

the long-chain non-coding RNA-H19 inhibits mTOR signaling pathway to block tumor cell proliferation and inhibit

Tumor cell growth;

the tumor cells are GH3 cells of subcutaneous transplantation tumor model and rat lactalbumin tumor in-situ induction growth

A cell;

the long-chain non-coding RNA-H19 inhibits the phosphorylation of the intracellular mTOR downstream substrate 4E-BP1

The level further inhibits mTOR function, thus realizing the inhibition of pituitary tumor and pituitary tumor cell growth.

2. The use according to claim 1, wherein said long non-coding RNA-H19

Inhibiting the proliferation capacity of GH3 cells, affecting the clone growth of GH3 cells, and inhibiting pituitary tumor GH3 cells in vivo

And (5) growing outside.