CN109722435B - Islet long-chain non-coding RNA1810019D21Rik and application thereof - Google Patents

Abstract

The present invention relates to islet long-chain non-coding RNA 1810019D21Rik and its application. The sequence of islet long non-coding RNA 1810019D21Rik is shown in SEQ ID No. 1; its application is related to diabetes or islet cells. The islet long-chain non-coding RNA 1810019D21Rik of the present invention can be used for various purposes related to diabetes or islet cells, and has a good application prospect.

Description

Islet long-chain non-coding RNA1810019D21Rik and application thereof

Technical Field

The invention relates to long-chain non-coding RNA1810019D21Rik of pancreatic islets and application thereof, in particular to regulation and control of the function of pancreatic beta cells by the RNA, belonging to the fields of cell biology and biomedicine.

Background

Long non-coding RNA (lncRNA) is a small RNA with transcript length more than 200bp, is positioned in nucleus or cytoplasm, is generally transcribed in eukaryotic cells and does not have protein coding function^[1-2]. lncRNA is expressed in lower abundance but with greater tissue and cellular expression specificity compared to protein-encoding mRNA. IncRNAs have different subcellular localization^[3]And play different roles in different cellular localisation, especially in the nucleus. Although lncRNA does not code protein, the lncRNA can be combined with specific protein to change the cytoplasmic localization of the protein, meanwhile, lncRNA can be used as a precursor series small molecule of miRNA and piRNA and participates in the regulation and control of protein coding genes from the transcription and post-transcription level, and a large amount of documents report that abnormally expressed lncRNA can cause various human diseases, such as skin diseases^[4]Coronary artery disease^[5]And diabetes mellitus^[6-7]。

An important function of the islet beta cells is the synthesis, storage and secretion of insulin and glucagon, which together regulate the blood glucose balance, which plays a key role in the pathogenesis of diabetes^[8]. In recent years, over 1100 islet lncRNA have been discovered in human islet tissue^[9-11]. The Kallen AN study found that lncRNA H19 wasThe combination of the beta-cell and the let-7 leads the beta-cell of the islet to generate insulin resistance^[12]。Motterle A^[13]lncRNAs, blinc2 and blinc3 are found to be highly expressed in islet beta cells, and have the characteristic of islet specific expression compared with lncRNA in islet tissues of other tissues. The results all indicate that the lncRNA participates in the function regulation of the islet beta cells and has important influence on the occurrence and development of the diabetes mellitus of human beings.

Research on lncRNA has attracted much attention in recent years, and research on lncRNA associated with pancreatic islet β cells is very limited. The method is used for exploring lncRNA which plays an important role in pancreatic islet beta cells, can reveal an important mechanism for regulating the number and the function of the pancreatic islet beta cells, provides a new biological target for diagnosing diabetes, and is an important scientific problem to be exploited in the research of the diabetes.

The Chinese invention patents of patent No. CN201410260768.2 and No. CN104059887B disclose the application of long-chain non-coding RNA Ovol2-AS, the GenBank accession number of the long-chain non-coding RNA Ovol2-AS is AK007144, and the long-chain non-coding RNA has the function of regulating the proliferation and the function of pancreatic beta cells. The patent is the existing research result of the lncRNA related to the islet beta cells, and more research is needed in the future to reveal the lncRNA related to the islet beta cells.

Disclosure of Invention

The invention aims to: overcoming the problems in the prior art, the islet long-chain non-coding RNA1810019D21Rik is provided and is used for the related application of diabetes mellitus or islet cells.

The technical scheme of the invention is as follows:

the islet long-chain non-coding RNA1810019D21Rik is characterized in that the sequence is shown in SEQ ID No. 1.

An expression vector comprising long non-coding RNA1810019D21Rik of islets as described above.

A host cell comprising an expression vector as described above.

The use of the long-chain non-coding RNA1810019D21Rik of pancreatic islets as described above for preparing or screening a medicament or pharmaceutical composition capable of increasing or reducing the expression level of the long-chain non-coding RNA1810019D21Rik of pancreatic islets.

Use of the long-chain non-coding RNA1810019D21Rik of pancreatic islets as described above for preparing or screening a medicament or pharmaceutical composition for treating diabetes.

The use of the long-chain non-coding RNA1810019D21Rik of pancreatic islets as described in the foregoing for preparing or screening a medicament or pharmaceutical composition for regulating insulin synthesis by pancreatic islet cells. The islet cells are islet beta cells.

Use of the long-chain non-coding RNA1810019D21Rik of pancreatic islets as described above for preparing or screening a product for diagnosing diabetes. The product is a preparation, a chip, a reagent or a kit.

Use of the long non-coding RNA1810019D21Rik of pancreatic islets as described hereinbefore for screening a method for diagnosing diabetes.

The inventor finds in practical research that the long-chain non-coding RNA1810019D21Rik of the pancreatic island has the function of regulating the synthesis of insulin by the beta cells of the pancreatic island as one of lncRNA which is differentially expressed in pancreatic island tissues of different diabetes model mice; the inventor discovers that the insulin synthesis can be remarkably promoted when the insulin is overexpressed after the long-chain non-coding RNA1810019D21Rik of the pancreatic island is overexpressed or knocked out in a min6 cell of a mouse pancreatic island beta cell line; in addition, the expression level of the islet long-chain non-coding RNA1810019D21Rik in the serum of a diabetic patient is remarkably lower than that of the serum of a normal person, which indicates that the expression level of the islet long-chain non-coding RNA1810019D21Rik in the serum of the diabetic patient is remarkably reduced. Through a series of researches, the inventor finally obtains the technical scheme of the invention.

The long-chain non-coding RNA1810019D21Rik of the pancreatic island can be used for multiple purposes related to diabetes or pancreatic island cells, and has good application prospect.

Drawings

FIG. 1 is a diagram illustrating the results of different models of mice and wild-type mice in the present invention, in which islet tissues together differentially express islet lncRNA. Wherein, con represents a wild type mouse.

FIG. 2 is a schematic diagram of the results of verifying the differential expression levels of islet lncRNA of different model mice and wild type mice in the embodiment of the present invention. Wherein, con represents a wild type mouse.

FIG. 3 is a diagram showing the results of the study on the expression specificity of lncRNA 1810019D21Rik in different tissues in the example of the present invention.

FIG. 4 is a graph showing the results of the localization study of IncRNA 1810019D21Rik in min6 cells in the example of the present invention.

FIG. 5 is a diagram showing the results of verifying the transfection efficiency of IncRNA 1810019D21Rik in min6 cell line in the example of the present invention.

FIG. 6 is a schematic diagram of the results of qRT-PCR detection of insulin synthesis 48 hours after lncRNA 1810019D21Rik is overexpressed or knocked out in the example of the present invention.

FIG. 7 is a diagram illustrating the positive expression result of gene insulin after overexpression or knock-out of lncRNA 1810019D21Rik in confocal detection according to an embodiment of the present invention.

FIG. 8 is a graph showing the results of the difference in lncRNA 1810019D21Rik expression in the human serum samples of diabetic and normal human by qRT-PCR in the examples of the present invention.

Detailed Description

The invention is described in further detail below with reference to embodiments and with reference to the drawings. The invention is not limited to the examples given.

Examples

First, main experimental materials

Mice: 5 weeks old C57BL/6J mice (wild type mice), 8 weeks old lepr^db/dbMice (diabetes model mice), 8 weeks old lep^ob/obMice (obese model mice), and 8-week-old HFD mice (high fat diet-induced obese model mice).

The main reagents are as follows: h BETA SS (10X) (Invitrogen), collagenase V (Sigma), Trizol lysate (Invitrogen), Reasy plus systemic mini kit (Qiagen), Ribo Zero Magnetic Gold kit (MRZG126, Illumina).

Second, Experimental methods

1. Extraction of islet tissue from different model mice

The mice were sacrificed by removing the cervical vertebrae, placed under a stereomicroscope, fixed with an adhesive tape, and a transverse incision was made under the xiphoid process with as large an opening as possible to expose the liver, stomach, duodenum and spleen. The forceps pick up the duodenum and turn it over downward, exposing the large papilla (white bulge) of the duodenum and gallbladder. The spleen was clamped up to expose the pancreas tail sufficiently to avoid pressure affecting perfusion. The Y-shaped triangular area where the bile duct, the hepatic duct and the pancreatic duct are intersected is found downwards from the gallbladder. The Y-shaped region was ligated once near the pancreas and ligated again 2mm above the ligation to prevent leakage during perfusion. The common duct of pancreas is tied at the large papilla of duodenum, and the common duct of pancreas is not tied at first, so that a space for inserting the needle is reserved. Sucking 2mL of the prepared collagenase V by using a 2.5mL syringe, inserting a needle from a duodenal tube, puncturing a duodenal papilla from the intestinal wall, slightly inserting into a common pancreatic duct by about 0.8-1cm, keeping the needle straight, tightening a ligature of the common pancreatic duct, tightening the needle together with the pancreatic duct, and preventing backflow. One person slightly pushes the syringe piston to inject collagenase into the pancreatic duct, and the other person pulls the ligature at the needle insertion position with both hands to prevent liquid backflow. At this point the pancreas should gradually expand and the pancreas tail begins to fill. After the injection is completed, the pancreatic envelope is gently separated from the pancreatic tail. The pancreas, which was associated with the stomach, duodenum, and small intestine, was gently separated and placed in 3mL of collagenase V solution (using a 50mL centrifuge tube as a container). Digestion was stopped at 37 ℃ for 28min at 60rpm to a uniform fine sand (particles less than 1 cubic mm). Digestion was stopped by adding 5mL of Hanks solution containing 10% fetal bovine serum. Poured into a 10cm dish, manually picked under an inverted microscope, placed in Hanks' solution, and held in a 10mL tube. Centrifuging and washing the precipitate for 2 times by Hanks liquid at 1000rpm and 4 ℃ for 2min, and taking the precipitate. The pellet was resuspended in 4mL of 25% Ficoll solution (10 mL tube was used as container) and 23%, 21%, 11% Ficoll solution, 2mL each, was slowly and uniformly added along the tube wall. In which case there should be a sharp interface between the concentrations. Centrifuge at 800g for 20min at 4 ℃. The Ficoll solution was aspirated, and islets were mainly concentrated in Ficoll at 11% and 21% concentration, with a small number of larger islets in 23% Ficoll. Washing with Hanks liquid for 2 times, re-suspending the islets with an appropriate amount of Hanks liquid, pouring into a 10cm dish again for fine picking and placing into an appropriate container, and then obtaining the purer islets.

2. Pancreatic islet tissue RNA extraction

Different mouse islet tissues were extracted as required by the Rneasy plus systemic mini kit (Qiagen) kit:

(1) to the sample was added 900. mu.l aiazol lysine reagent, allowed to stand at room temperature for 5min, and 100. mu.l gDNA Elimator solution was added and mixed vigorously for 15 s.

(2) Adding 180 μ L of chloroform, violently mixing by vortex for 15s, and standing at room temperature for 2-3 min.

(3) Centrifuge at 12000g for 15min at 4 ℃ and aspirate the supernatant.

(4) Add 1 volume (supernatant volume) of 70% ethanol and mix well.

(5) The liquid is added into an adsorption column of a mini spin gum collecting pipe in times, the maximum volume of each time is 700 mu L, and the liquid is centrifuged for 1min at room temperature of 8000 g.

(6) mu.L of Buffer RWT was added to the adsorption column and centrifuged at 8000g for 1min at room temperature.

(7) Add 500. mu.L Buffer rpe to the adsorption column and centrifuge at 8000g for 1min at room temperature, and repeat once.

(8) And (3) putting the adsorption column into a new collecting pipe, and adding 30 mu L of RNase-free water for elution to obtain the total RNA of the pancreatic islet tissue.

3. Screening for differential genes by RNA-seq analysis

The obtained total RNA passes through a kit Ribo Zero Magnetic Gold kit (MRZG126, Illumina) to denucleate a ribosome, the Illumina HiSeq2000 is used for building a library, and Cufflinks (v2.2.1) is used for obtaining a reads value of each sample so as to judge whether each lncRNA is a new lncRNA or not, and the transcript also has a code 'j, i, o, u, x', the length is more than 200bp, and the transcript can be compared in a nonode database to be known lncRNA; the other transcripts were used to predict whether they encoded proteins or not using Coding Potential calcium μ late (CPC) and TransDecode, transcripts that did not encode proteins were defined as new lncRNA, FPKMs were calculated for each set of samples using Cuffdiff (v2.2.1), transcripts satisfying p value < 0.05 were defined as differentially expressed genes, and log2(fold change) <1 was a significantly differentially expressed gene.

Compared with the wild mouse islet tissue, a great amount of different islet lncrnas were screened from both the obese and diabetic model mouse islet tissues, but only 8 islet lncrnas (TCONS _00033309, TCONS _00035452, TCONS _00042278TCONS _00057502, TCONS _00091281, TCONS _00115452, and TCONS _00117141TCONS _00117153) were significantly reduced in all model mice compared with the wild mouse islet tissue (as shown in fig. 1).

4. Differential validation of lncRNA expression

And (3) synthesizing specific qRT-PCR primers by using the lncRNA with higher abundance and large expression difference, and verifying the expression difference. The similarity between the verification result and the sequencing result is 80%, which indicates that the sequencing result is more accurate (as shown in FIG. 2).

5. min6 separation of nuclear and cytoplasmic substances

(1)1×10⁶The min6 cells were added to 300. mu.L of ice-cold Cell Fractionation Buffer (PARIS)^TMKit, life) was left standing at room temperature for 5 min.

(2) Centrifuge at 500 Xg for 5min at 4 ℃. Transfer supernatant to a new 2mL rnase-free tube.

(3) Adding 300 mu L of ice-cold Cell precipitation Buffer into the precipitate, violently mixing, adding equal volume of 2x lysine and equal volume of absolute ethyl alcohol into all samples, transferring the mixed solution into an adsorption column, and centrifuging for 1min at 1000 g.

(4) Add 700. mu.L wash solution 1 and centrifuge at 1000g for 1 min.

(5) Wash twice with 500. mu.L wash solution 2/3 and centrifuge at 1000g for 1 min.

(6) Finally, 30. mu.L of solution Elution RNA preheated to 95 ℃ was added to obtain nuclear and cytoplasmic RNA, respectively.

6. Construction of lncRNA 1810019D21Rik (i.e., lncRNA TCONS-00115452) expression vector

(1) The full length of the lncRNA 1810019D21Rik-CDS region is inquired in NC BETA I, and the sequence is shown as SEQ ID No. 1.

(2) Synthetic lncRNA 1810019D21Rik-CDS upstream and downstream primers were designed by Primer 5.

(3) The optimal annealing temperature of lncRNA 1810019D21Rik was examined using islet tissue as template according to the high fidelity enzyme kit instructions (2x planta max Buffer 5. mu.L, 10. mu.M DNTP Mix 0.2. mu.L, template 1. mu.L, upstream and downstream primers 0.4. mu.L each, DNA polynerase 0.2. mu.L).

(4) The PCR products were separated on a 1% agarose gel at 200V and the band of interest was excised under UV light.

(5) DNA was recovered according to Tian Gen kit instructions:

column balancing: adding 500 μ L of equilibrium liquid BETA into adsorption column CA1 (placing adsorption column into collection tube), centrifuging at 13400 × g for 1min, pouring off waste liquid in the collection tube, and placing adsorption column back into the collection tube.

② adding equal volume of solution PN into the rubber block, placing in 50 ℃ water bath, and turning the centrifuge tube up and down gently to ensure the rubber block to be fully dissolved.

③ adding the solution obtained in the previous step into an adsorption column CA1 (the adsorption column is placed into a collecting pipe), standing for 2min at room temperature, centrifuging for 1min at 13400 Xg, pouring the waste liquid in the collecting pipe, and placing the adsorption column into the collecting pipe again.

Fourthly, 600 mul of rinsing liquid PW is added into the adsorption column CA1, 13400 Xg is centrifuged for 1min, waste liquid in the collecting pipe is poured out, and the adsorption column is put into the collecting pipe again.

Fifthly, putting the centrifugal adsorption column CA1 back into the collecting pipe, centrifuging for 2min at 13400 Xg, and removing the rinsing liquid as much as possible. The column was left at room temperature for several minutes.

Sixthly, putting the adsorption column CA1 into a clean centrifugal tube, hanging and dripping a proper amount of elution buffer solution E BETA to the middle position of the adsorption film, and standing for 2min at room temperature. The DNA solution was collected by centrifugation at 13400 Xg for 2 min.

(6) The recovered DNA plasmid was cut into cohesive ends using xho1 and BETA amH1, ligated with plasmid pLVX-IRES-ZsGreen1, transfected into competent E.coli DH5a, and single clone sequencing was performed to obtain an expression vector of lncRNA 1810019D21 Rik.

7. lncRNA 1810019D21Rik expression vector transfection

Min6 cells were transiently transfected with lncRNA 1810019D21Rik siRNA, lncRNA 1810019D21Rik CDS with lipo2000, control NC and empty plasmid plvx-IRES zsgreen1, and transfection efficiency was judged by green fluorescence (as shown in FIG. 5 b).

8. Confocal detection of positive expression quantity of gene insulin

(1) Min6 cells overexpressing or knocking-out lncRNA 1810019D21Rik were fixed with 4% cold poly methanol for 20min after 48 h.

(2) PBST was washed 3 times for 10min each, and 0.2% Triton X-100 solution was allowed to permeate for 20min at room temperature.

(3) PBST was washed 3 times for 10min each, and blocked at 37 ℃ for 2 h.

(4) PBST was washed 3 times for 10min each, primary antibody incubated, and incubated overnight at 4 ℃.

(5) PBST was washed 3 times for 10min each, and a secondary antibody was incubated at 37 ℃ for 1 h.

(6) PBST was washed 3 times for 10min each and incubated 5min at 37 ℃ with 5. mu.M DAPI in the dark.

(7) PBST was washed 3 times for 10min each time, and positive expression of gene insulin was observed with Zeiss microscope at a magnification of 40 times.

Third, research results

3.1 screening for differentially expressed IncRNAs

Differential lncRNA of islet tissues of all model mice is analyzed through Cuffdiff (v2.2.1), the expression quantity of 8 lncRNA is found to be remarkably reduced in obese and diabetic mice (shown in figure 1), and the similarity between the verification result and the sequencing data is about 80% through verifying the sequencing data through qRT-PCR (quantitative reverse transcription-polymerase chain reaction), which indicates that the sequencing data is accurate and reliable (shown in figure 2). The lncRNA 1810019D21Rik (i.e. lncRNA TCONS-00115452) is abundantly expressed in mouse islet tissues and has larger variation.

3.2 IncRNA 1810019D21Rik tissue-specific examination

RNA of heart, liver, spleen, lung, kidney, brain, white fat, palm fat, muscle, pancreatic islet and exocrine cell of the mouse is extracted, expression difference of the RNA in different tissues is detected through qRT-PCR, and the result shows that the RNA is highly expressed in the pancreatic islet, exocrine cell, liver and muscle, wherein the expression level of the RNA in the pancreatic islet tissue is six times that of the exocrine cell, which indicates that the lncRNA 1810019D21Rik has tissue specificity (shown in figure 3).

3.3 cellular localization of lncRNA 1810019D21Rik

The nucleus and cytoplasm of min6 cells were separated by nucleoplasm separation kit, and the localization of lncRNA 1810019D21Rik was detected by qRT-PCR, U6 and GAPDH were used as control for cytoplasm and nucleus, respectively. The results showed that lncRNA 1810019D21Rik is mainly located in the nucleus (as shown in FIG. 4).

3.4 verification of transfection efficiency of IncRNA 1810019D21Rik

The cell was transfected with lncRNA 1810019D21Rik lentivirus and control lentivirus plvx-ireszsgreen 1 stably in min6, lncRNA 1810019D21Rik siRNA was transiently transfected, NC was used as control, RNA was harvested after transfection for 48h, and the expression level was detected by qRT-PCR, which showed that the expression level was significantly increased when lncRNA 1810019D21Rik was overexpressed, while the knockout efficiency was about 70% when lncRNA 1810019D21Rik was knocked out (as shown in FIG. 5 a), and FIG. 5b shows that lncRNA 1810019D21Rik green fluorescence expression was overexpressed in min6 cells, thus proving that the transfection efficiency was high.

3.5lncRNA 1810019D21Rik regulates insulin Synthesis

After lncRNA 1810019D21Rik is overexpressed or knocked-out, synthesis of insulin in min6 cells is detected, and the result shows that when lncRNA 1810019D21Rik is overexpressed, the expression levels of genes ins1 and ins2 are obviously increased, and when lncRNA 1810019D21Rik is knocked-out, the expression levels of genes ins1 and ins2 are reduced, which indicates that lncRNA 1810019D21Rik promotes islet transcription (as shown in fig. 6).

3.6 confocal detection of IncRNA 1810019D21Rik regulating insulin Synthesis

The lncRNA 1810019D21Rik is overexpressed or knocked out in min6 cells, the sinulin and DAPI are co-stained, the positive rate of the gene insulin is observed by a Zeiss microscope, and the result shows that the expression level of the gene insulin is remarkably increased by overexpressing the lncRNA 1810019D21Rik compared with the control group (as shown in a figure 7).

3.7qRT-PCR detection of expression differences of lncRNA 1810019D21Rik in human serum samples from diabetes and Normal

Human Serum samples were extracted with miRNeasy Serum/Plasma Kit (Qiagen) for total RNA, 2.5. mu.l of control C.elegans spiked-in control cell-miR-39 (Applied BETA-biosystems) was added, and finally eluted with 20. mu.l of RNase-free water. The expression difference of lncRNA 1810019D21Rik was detected by qRT-PCR, and the result showed that lncRNA 1810019D21Rik was significantly reduced in the serum sample of diabetic patients compared to the normal human serum sample (as shown in fig. 8).

The research shows that the islet long-chain non-coding RNA1810019D21Rik (namely lncRNA 1810019D21Rik) can be used for regulating and controlling islet beta cells to synthesize insulin, is used as a new biological index for diagnosing diabetes, is used for preparing or screening a medicament or a medicinal composition for treating diabetes, and is used for preparing or screening a medicament or a medicinal composition capable of improving or reducing the expression level of the islet long-chain non-coding RNA1810019D21 Rik.

According to the invention, the differentially expressed islet lncRNA in islet tissues of different model mice is screened through RNA-seq, the significant reduction of the expression level of lncRNA 1810019D21Rik in obesity and diabetes model mice is verified, the function of regulating islet beta cells by the lncRNA is researched, the lncRNA is found to promote the synthesis of insulin for the first time, and the expression level of the lncRNA in a serum sample of a diabetes patient is found to be significantly reduced compared with that of a normal person, so that abundant resources are provided for the continuous research of the function and mechanism of the islet lncRNA related to diabetes, a foundation is laid for further screening and exploring the new function and related regulation mechanism of the islet lncRNA, and a new biological index is provided for the diagnosis and treatment of diabetes.

Reference to the literature

[1]Fatica A,Bozzoni I.Long non-coding RNAs:new players in cell differentiation and development[J].Nat.Rev.Genet.,2014,15(1):7-21.

[2]Kopp F,Mendell JT.Functional Classification and Experimental Dissection of Long Noncoding RNAs[J].Cell,2018,172(3):393-407.

[3]Derrien T,Johnson R,Bussotti G et al.The GENCODE v7catalog of human long noncoding RNAs:analysis of their gene structure,evolution,and expression[J].Genome Res.,2012,22(9):1775-1789.

[4]Szell M,Danis J,Bata-Csorgo Z et al.PRINS,a primate-specific long non-coding RNA,plays a role in the keratinocyte stress response and psoriasis pathogenesis[J].Pflugers Arch.,2016,468(6):935-943.

[5]Yang Y,Cai Y,Wu G et al.Plasma long non-coding RNA,CoroMarker, a novel biomarker for diagnosis of coronary artery disease[J].Clin.Sci.(Lond.), 2015,129(8):675-685.

[6]Kaur S,Mirza AH,Brorsson CA et al.The genetic and regμlatory architecture of ERBB3-type 1diabetes susceptibility locus[J].Mol.Cell. Endocrinol.,2016,419:83-91.

[7]Wang M,Wang S,Yao D et al.A novel long non-coding RNA CYP4B1-PS1-001regμlates proliferation and fibrosis in diabetic nephropathy[J]. Mol.Cell.Endocrinol.,2016,426:136-145.

[8]Ouedraogo R,Naslund E,Kirchgessner AL.Glucose regμlates the release of orexin-a from the endocrine pancreas[J].Diabetes,2003,52(1):111-117.

[9]Benner C,van der Meμlen T,Caceres E et al.The transcriptional landscape of mouse beta cells compared to human beta cells reveals notable species differences in long non-coding RNA and protein-coding gene expression[J]. BMC Genomics,2014,15:620.

[10]Ku GM,Kim H,Vaughn IW et al.Research resource:RNA-Seq reveals unique features of the pancreatic beta-cell transcriptome[J].Mol.Endocrinol.,2012, 26(10):1783-1792.

[11]Moran I,Akerman I,van de Bunt M et al.Human beta cell transcriptome analysis uncovers lncRNAs that are tissue-specific,dynamically regμlated,and abnormally expressed in type 2 diabetes[J].Cell Metab.,2012, 16(4):435-448.

[12]Kallen AN,Zhou XB,Xu J et al.The imprinted H19 lncRNA antagonizes let-7 microRNAs[J].Mol.Cell,2013,52(1):101-112.

[13]Motterle A,Gattesco S,Peyot ML et al.Identification of islet-enriched long non-coding RNAs contributing to beta-cell failure in type 2 diabetes[J]. Molecμlar metabolism,2017,6(11):1407-1418。

Sequence listing

<110> university of Chinese pharmacy

<120> islet long-chain non-coding RNA1810019D21Rik and application thereof

<160> 1

<170> SIPOSequenceListing 1.0

<210> 1

<211> 1647

<212> DNA/RNA

<213> mouse (Mus musculus)

<400> 1

caaggacaga gcttcatact tggccactcg cagcagcctg agctttgacc cacaccccat 60

cgaccaactg gcacagaact gtttgctcgt agaggggggt gggtggcact ctgaaggcta 120

tccagcacga tccggccctt tcccttggca attctgtgac actttccaat ctaaggccga 180

cagagcaagg caaccagatt tttttctaca gcaccctccc aggtgagcag tgggcctgga 240

gcccttctgt aacctggcat ctgtaggtct tctcattaag aacgagagtc agaagaggaa 300

aggacttaac actcattaca actcagtcat ggggctctga agcgtcccga ggtcccctct 360

ttcccaccct acctgtcggc tgcgcggctc caccacttgc caaactagga ggattaagtc 420

ggtctcgtcc gagcctaggt ccggccccag cgcaccggca gtggccccga acaaaaccac 480

cagcgactga gcaggcactc tgagtcctcc gcaaaagaat gtgcaggatc agaggcctct 540

ttccattctg caagggtcac gcaagaccca tggaattcaa caggaggcaa tctagggact 600

ataaaaggtc gcgacgtctg gcttgaggtt aatggaagtt acaaagggtc aaaagccgac 660

agtgagcgct accaggtgtc gggtcggcgt agtgccggca gtgcaaggcg cacgttactg 720

tggacctgaa ggctcaggcc ttggtgccaa ggccgccctg tgggctcagg atagtttccc 780

acctactcga ctgtaattag tactctggcg agagtaaaaa ggtactggcg tgggacttag 840

aatacaggat gccaccggct gaagaagaac cttgtgtgtt tggaaaagcg ggctgcttcc 900

tatcggtctt ggataaaagg caacccagag cccgggtgca tccctaactc atgcccaagg 960

aaggtcgtgc ggagccagag gacctagaat gtatagtgcg ccttctgggt ttgtccacac 1020

aatagcgcag gaacacagcg acccaatata gtgagaagtg gccgcagcgc acaccctgag 1080

acccttcacc aggcggccca cccggagcct gcctactcag ccccgctacc acgataagcc 1140

tggaatgtca aatatttgtt ctaggaaccc agtgaaacct tgaggtaaag cccctcccct 1200

cctccctccc attccggagc gcgcctgcgt cagtccaggc tttccggtaa ggacactccc 1260

ttccaactag ttccttctcg acacccctgc aacccatcgt cctgactatt taaacttcac 1320

aaggtcttcg ctacaggctc aggttcagtg aggcagagtc ttggcttccc tacccagtta 1380

tcccagagct actggtatcg gtttctaccc acaacctccg agcaaggctc ccctttcaac 1440

ctgtagacct cgcctcccga ttctcgggag atcttgctga ctgtggaatc cctggctgtt 1500

ccgaaccccc tggaaagaag cggctgttcc caggtttctg caaggtaacg aatctgaaga 1560

gtcaacagaa ttgagcggct ccattcattc gactctggtt ctctcaacca ggcattcccc 1620

atattaaaac aattccagaa cttcaaa 1647

Claims (4)

1. The application of the islet long-chain non-coding RNA1810019D21Rik in preparing the medicines or the medicine compositions for treating diabetes mellitus is disclosed, wherein the sequence of the islet long-chain non-coding RNA1810019D21Rik is shown as SEQ ID No. 1.

2. The application of the long-chain non-coding RNA1810019D21Rik of the pancreatic islets in preparing the medicine or the pharmaceutical composition for regulating the synthesis of insulin by the pancreatic islet cells, wherein the pancreatic islet cells are pancreatic islet beta cells, and the sequence of the long-chain non-coding RNA1810019D21Rik of the pancreatic islets is shown as SEQ ID No. 1.

3. The application of the reagent for detecting the long-chain non-coding RNA1810019D21Rik of the pancreatic islets in preparing the product for diagnosing the diabetes mellitus is disclosed, wherein the sequence of the long-chain non-coding RNA1810019D21Rik of the pancreatic islets is shown as SEQ ID No. 1.

4. Use according to claim 3, wherein the product is a preparation or a chip.

Images