CN111647659A

CN111647659A - New application of LOC285627

Info

Publication number: CN111647659A
Application number: CN202010617801.8A
Authority: CN
Inventors: 杨承刚; 孙耀兰
Original assignee: Beijing Medintell Bioinformatic Technology Co Ltd
Current assignee: Beijing Medintell Bioinformatic Technology Co Ltd
Priority date: 2020-06-30
Filing date: 2020-06-30
Publication date: 2020-09-11

Abstract

The invention discloses a new application of LOC285627, and particularly discloses an application of LOC285627 in diagnosis and treatment of laryngeal squamous cell carcinoma. The invention also discloses a tool for diagnosing the laryngeal squamous cell carcinoma, a method for screening the medicament for treating the laryngeal squamous cell carcinoma, and a medicament for treating the laryngeal squamous cell carcinoma.

Description

New application of LOC285627

Technical Field

The invention belongs to the field of biomedicine, and relates to a new application of LOC 285627.

Background

Laryngeal carcinoma is a common malignant tumor of the head and neck, accounts for the second place of the head and neck tumor, the incidence rate of the laryngeal carcinoma accounts for about 5 percent of the malignant tumor of the whole body, the squamous cell carcinoma accounts for about 93 to 99 percent of the total number of laryngeal carcinoma, and the laryngeal squamous cell carcinoma mostly originates from vocal cords, accounting for about 60 percent. Retrospective analysis is carried out on 306 laryngeal malignant tumors by using xanthoxinhui and the like, and the squamous cell carcinoma accounts for 99.7 percent of all malignant tumors, wherein high and medium squamous cell carcinoma is mainly used; the primary site of laryngeal carcinoma is mostly glottic, and secondly supraglottic and subglottic; smoking and drinking are closely related to the occurrence of tumors. Although the pain of patients can be relieved to different degrees by operation, radiotherapy and chemotherapy, dysfunction and local deformity are often caused after treatment, and the recurrence rate is high, thus seriously threatening the physical and psychological health of patients. The second mutation hypothesis proposed by Knudson in 1971 is a scientific prophetic for the tumorigenesis mechanism, widely accepted by researchers in tumor genetics, and has led more than 30 years of research into the molecular mechanisms of tumorigenesis. The study of different tumor related genes is the basic premise for understanding the molecular mechanism of tumorigenesis, developing gene diagnosis and gene therapy, and the cloning and localization of disease-causing genes including tumors are still an important content after the completion of human genome planning. Srinivus and the like find that the tumor can be accompanied with the change of a tumor marker at an early stage, and the detection of the content change of the tumor marker is beneficial to the early discovery of the tumor. Gene level research work will be key to the eradication of all tumors, including laryngeal cancer. Therefore, the search and cloning of the laryngeal cancer specific related gene provides a wide prospect for research on the molecular mechanism, gene diagnosis and gene therapy of laryngeal cancer.

Although the means for tumor diagnosis and treatment are changing day by day, the overall survival rate of laryngeal cancer is not improved significantly worldwide, especially for patients in advanced stages. The reason is mainly shown in three aspects: firstly, the early clinical manifestations of laryngeal cancer are various, great difficulty is brought to early diagnosis of laryngeal cancer, most patients cannot receive timely and effective treatment in early stage, and the key for improving the early diagnosis rate of laryngeal cancer is how to find the molecular marker for early diagnosis of laryngeal cancer. Secondly, the treatment of the laryngeal cancer is mainly performed by operation, radiotherapy and chemotherapy are assisted, and although the ratio of partial laryngeal resection and early glottic laryngeal cancer treatment by C02 laser is improved, the laryngeal function is completely or partially preserved, and postoperative complications are reduced, so that the quality of life of a patient is improved, the postoperative recurrence rate is still high, if a laryngeal cancer tumor promotion gene or a tumor inhibition factor can be found out, a potential drug action target point is screened out, and a foundation is laid for developing a specific vaccine or/and a new anticancer drug related to the laryngeal cancer. Thirdly, different patients have different laryngeal cancer in the aspects of pathological type, tumor differentiation degree, lesion severity degree in treatment and the like, different laryngeal cancer patients have different clinical manifestations and physical conditions, and have different sensitivities to postoperative radiotherapy and chemotherapy, and how to find a molecular marker for curative effect monitoring and prognosis judgment lays a foundation for realizing personalized treatment of laryngeal cancer.

Disclosure of Invention

The invention aims to provide a long-chain non-coding RNA marker for diagnosing laryngeal squamous cell carcinoma. The invention utilizes QPCR experiments to prove that the expression level of LOC285627 in cancer tissues of laryngeal squamous cell carcinoma patients is obviously lower than that of tissues beside cancer, so that LOC285627 can be used as a molecular marker for diagnosing laryngeal squamous cell carcinoma.

In order to test the purpose, the invention adopts the following technical scheme:

the invention provides an application of a reagent for detecting long-chain non-coding RNA expression in preparing a laryngeal squamous cell carcinoma diagnosis product. The long non-coding RNA is LOC 285627.

Further, the reagent includes a reagent used for detecting the expression level of LOC285627 by a reverse transcription PCR method, a real-time quantitative PCR method, a chip detection method, a southern blotting method, a northern blotting method or an in situ hybridization method.

Further, the reagent comprises an amplification primer used for detecting the expression level of LOC 285627.

In a specific embodiment of the invention, the amplification primer sequences are shown as SEQ ID NO.1 and SEQ ID NO. 2.

The invention provides a product for diagnosing laryngeal squamous cell carcinoma, which diagnoses laryngeal squamous cell carcinoma by detecting the expression of the long-chain non-coding RNA in a sample.

Further, the product includes but is not limited to a chip, a kit, a strip, or a high throughput sequencing platform; the high-throughput sequencing platform is a special diagnostic tool, and with the development of high-throughput sequencing technology, the construction of an RNA expression profile of a person becomes very convenient work. By comparing the RNA expression profiles of patients with disease and normal populations, it is easy to identify which RNA abnormalities are associated with disease. Therefore, the application of LOC285627 in high-throughput sequencing to know that the expression abnormality of LOC285627 is related to laryngeal squamous cell carcinoma also belongs to the application of LOC285627, and is also within the protection scope of the invention.

The kit comprises a reagent for detecting the expression level of LOC285627, wherein the reagent comprises nucleic acid combined with LOC285627 or a DNA sequence thereof, and the nucleic acid comprises primers and/or probes used when SYBR Green, TaqMan probes, molecular beacons, double-hybridization probes or composite probes are used for detecting the expression level of LOC 285627.

The chip comprises a reagent for detecting the expression level of LOC285627, wherein the reagent comprises a nucleic acid combined with LOC285627 or a DNA sequence thereof, and the nucleic acid comprises a primer and/or a probe capable of detecting the expression level of LOC 285627.

The test paper comprises a reagent for detecting the expression level of LOC285627, wherein the reagent comprises nucleic acid combined with LOC285627 or a DNA sequence thereof, and the nucleic acid comprises a primer and/or a probe capable of detecting the expression level of LOC 285627.

The kit may further comprise: container, instructions for use, positive control, negative control, buffer, adjuvant or solvent. For example, a solution for suspending or immobilizing cells, a detectable label or label, a solution for facilitating hybridization of nucleic acids, a solution for lysing cells, or a solution for nucleic acid purification.

The instruction book of the kit of the invention describes how to use the kit for detection, how to judge the tumor development by using the detection result and how to select the treatment scheme.

With the kit of the present invention, LOC285627 can be detected by various methods (including but not limited to) selected from the group consisting of: reverse transcription PCR, real-time quantitative PCR, chip detection, southern blotting, northern blotting, or in situ hybridization. The detection mode can be adjusted and changed by those skilled in the art according to actual conditions and needs.

Further, the primer sequences are shown as SEQ ID NO.1 and SEQ ID NO. 2.

The invention provides an application of LOC285627 in screening of a candidate drug for preventing or treating laryngeal squamous cell carcinoma.

Further, the steps of screening candidate drugs are as follows:

treating a system expressing or containing LOC285627 with a candidate substance; and

detecting the expression of LOC285627 in said system;

wherein, if the candidate substance can increase the expression of LOC285627, preferably significantly, the candidate substance is a candidate drug for preventing or treating laryngeal squamous cell carcinoma.

In the present invention, the system includes (but is not limited to): a cell system, a subcellular system, a solution system, a tissue system, an organ system, or an animal system. The candidate substances include (but are not limited to): nucleic acid molecules, carbohydrates, lipids, small molecule chemical drugs, antibody drugs, polypeptides, proteins, or vectors.

In the present invention, the steps further include: the obtained candidate drugs are subjected to further cell experiments and/or animal experiments to further select and identify drugs useful for preventing, alleviating or treating laryngeal squamous cell carcinoma from the candidate drugs.

The invention provides an application of LOC285627 in preparation of a medicine for preventing or treating laryngeal squamous cell carcinoma.

The medicament comprises a substance promoting LOC285627 expression.

The substance for promoting the expression of LOC285627 comprises an overexpression vector containing LOC285627 and a host cell containing the overexpression vector.

The invention provides a pharmaceutical composition for treating laryngeal squamous cell carcinoma, which comprises a substance for promoting LOC285627 expression and/or other pharmaceutically acceptable carriers compatible with the substance.

The pharmaceutically acceptable carriers include, but are not limited to, diluents, excipients, binders, wetting agents, absorption enhancers, surfactants, humectants, adsorptive carriers, lubricants, buffers, stabilizers, bacteriostats, isotonizing agents, chelating agents, pH control agents.

The pharmaceutical compositions of the present invention may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir. Oral administration or injection administration is preferred. The pharmaceutical composition of the present invention may contain any of the usual non-toxic pharmaceutically acceptable carriers, adjuvants or excipients.

The pharmaceutical composition of the invention can also be used in combination with other drugs for the treatment of squamous cell carcinoma of the larynx, and the other therapeutic compounds can be administered simultaneously with the main active ingredient, even in the same composition. Other therapeutic compounds may also be administered alone in a composition or dosage form different from the main active ingredient.

Preferably, it can be carried out by means of gene therapy. For example, a substance that promotes the expression of LOC285627 (e.g., an overexpression vector) can be directly administered to a subject by a method such as injection, and such techniques are well known to those skilled in the art.

The present invention also provides a method for diagnosing laryngeal squamous cell carcinoma, the method comprising the steps of:

(1) obtaining suspected diseased tissue of the throat of a subject;

(2) detecting the expression level of LOC285627 in the tissue of step 1;

(3) correlating the measured expression level of LOC285627 with the presence or absence of disease in the subject.

(4) When the expression level of LOC285627 in the suspected diseased tissue in the larynx of the subject is significantly reduced compared to the normal control, the subject is judged to have laryngeal squamous cell carcinoma, or the subject is judged to have a high risk of laryngeal squamous cell carcinoma, or a patient with laryngeal squamous cell carcinoma is judged to have a relapse, or a patient with laryngeal squamous cell carcinoma is judged to have a poor prognosis.

In the context of the present invention, a "normal control" may refer to a normal human not suffering from laryngeal squamous cell carcinoma, but also to the paracancerous tissues of a human suffering from laryngeal squamous cell carcinoma.

In the context of the present invention, "diagnosing" includes determining whether a subject has suffered a disease, determining whether a subject is at risk of suffering a disease, determining whether a patient has relapsed, determining the responsiveness of a patient to a drug treatment, or determining the prognosis of a patient.

Drawings

FIG. 1 shows a statistical chart of LncRNA overexpression.

Detailed Description

The present invention will be described in further detail with reference to the accompanying drawings and examples. The following examples are intended to illustrate the invention only and are not intended to limit the scope of the invention. Experimental procedures without specific conditions noted in the examples, generally following conventional conditions, such as Sambrook et al, molecular cloning: the conditions described in the laboratory Manual (New York: Cold Spring harbor laboratory Press,1989), or according to the manufacturer's recommendations.

Example 1 differential expression molecule validation

First, research object

Cancer tissues and corresponding paracancerous tissues were collected from 45 patients with laryngeal squamous cell carcinoma. Exclusion criteria were included: firstly, the patient has no primary tumor at other parts except the primary tumor, and the primary tumor is not transferred; ② there is no history of cardiovascular and cerebrovascular diseases such as diabetes, hypertension, etc.; thirdly, the history of infectious diseases such as hepatitis, syphilis, tuberculosis, HIV and the like is avoided; fourthly, no history of family hereditary diseases; before operation, radiotherapy, chemotherapy and biological treatment are not performed.

Taking sample precautions: when the tumor specimen is cut, the non-necrotic tumor tissue in the central area of the tumor is cut, the normal tissue beside the cancer is cut away from the tumor as far as possible and is more than 1.5cm away from the edge of the tumor, and the mucosal epithelial tissue is cut as far as possible to avoid the deep connective tissue.

Second, tissue total RNA extraction

Tissue total RNA was extracted using the Norgen RNA extraction kit:

tissue lysis

1) Weighing about 20mg of the isolated tissue sample in a clean area with less RNase interference by using a mortar containing a proper amount of liquid nitrogen, and grinding the sample into powder by using a pestle;

2) transferring the sample to a 2mL centrifuge tube without RNase;

3) adding 300uL lysine solution, placing in a homogenizer, and grinding for 1-5 min;

4)12000g, centrifuging for 10min at 4 ℃, transferring supernatant into a new centrifuge tube with the volume of 1.5 mL;

5) adding 600ul RNase-Free Water, and mixing with a vortex machine;

6) adding 20ul protease K, bathing at 55 deg.C for 15min, and continuously vortex and mixing;

7)14000g, centrifuging for 1min at room temperature to precipitate cell debris at the bottom of the centrifuge tube, taking supernatant and transferring the supernatant into another centrifuge tube without 1.5mL of RNase;

8) adding 450ul of 95% ethanol, and mixing by vortex;

RNA adsorption:

9) adding 650ul of lysate containing ethanol into a centrifugal column, and centrifuging for 1min at 14000 g;

10) abandoning the lower layer, and resetting the collecting pipe on the column;

11) repeating the steps from 9) to 10) according to the volume of the lysate;

12) adding 400ul Wash solution, 14000g, and centrifuging for 2 min;

13) abandoning the lower layer, and placing the column on a new collecting pipe;

and (3) DNase treatment:

14) adding 100ul enzyme incorporation Buffer and 15ul DNase I, and centrifuging for 1min at 14000 g;

15) moving the solution in the collecting pipe into the column again;

16) standing at room temperature for 15 min;

RNA washing:

17) adding 400ul Wash solution, centrifuging at 14000g for 1min, discarding the lower layer, and resetting the collection tube on the column;

18) adding 400ul Wash solution, 14000g, centrifuging for 2min, and discarding the collecting pipe;

RNA elution:

19) the column was placed in a 1.7mL Elution tube;

20) adding 30ul of Elution Buffer;

21) centrifuge at 200g for 2min to allow the solution to bind well to the column, and then centrifuge at 14000g for 1 min.

Third, reverse transcription

The reverse transcription kit (DDR037A) was purchased from Bao bioengineering (Dalian) Co., Ltd.

Taking the extracted total RNA (1 mu g) as a template, adding the following reaction system:

Buffer4μL，

RT Enzyme Mix 1. mu.L, Oligo dT Primer (50. mu.M) 1. mu.L, Random 6mers (100. mu.M) 1. mu.L, as RNase-free ddH₂O make up the reaction volume to 20. mu.L. The mixture was incubated at 37 ℃ for 15min and 85 ℃ for 5s to obtain cDNA. The cDNA can be used for IncRNA Real-time PCR detection.

IV, QPCR

SYBRPremix Ex Taq for fluorescent Real-time (Real-time) quantitative PCR (polymerase chain reaction)^TM(Tli RNaseH Plus) kit was manufactured by Takara, Japan. The operation was performed as per the instructions.

By using

And analyzing the expression level of LncRNA by a relative quantification method, wherein Ct is the intensity value of a fluorescence signal detected in a reaction system by a thermal cycler. The calculation method comprises the following steps: delta Ct (Ct target gene-Ct reference gene) cancer tissue experimental group- (Ct target gene-Ct reference gene) control tissue group,

the expression of the target gene in the experimental group is shown as the fold change relative to the control group, and the analysis of the experimental data is performed by the Bio-RAD analysis software.

Designing a primer: primers were designed by the Primer design tool of NCBI (Primer BLAST) based on LncRNA sequence as follows:

LOC285627

an upstream primer: 5'-AATCAAGTGGCTAAGTCA-3' (SEQ ID NO. 1); a downstream primer: 5'-GGATTCATCTCATTAACTCTG-3' (SEQ ID NO. 2).

LOC100506860

An upstream primer: 5'-CCACTGAGAAGTCCTAAG-3' (SEQ ID NO. 3); a downstream primer: 5'-TGATGATGAAACTGGCTTA-3' (SEQ ID NO. 4).

LOC388692

An upstream primer: 5'-CTCACAGTCGGACCTTAG-3' (SEQ ID NO. 5); a downstream primer: 5'-ACAGAACGGAAGACAAGA-3' (SEQ ID NO. 6).

Primers were designed based on the GAPDH (internal reference gene) sequence, the upstream primer: 5'-CTCTGGTAAAGTGGATATTGT-3' (SEQ ID NO. 7); 5'-GGTGGAATCATATTGGAACA-3' (SEQ ID NO. 8).

Fifth, statistical analysis

Statistical software SPSS19.0 is adopted for data analysis, and paired T test is used for judging whether the LncRNA expression in cancer tissues and cancer adjacent tissue samples has statistical difference. The statistical tests are bilateral tests, and the difference is statistically significant when P is less than 0.05.

Sixthly, the results

As shown in table 1, the expression levels of LOC285627, LOC100506860 and LOC388692 in the cancer tissues of laryngeal squamous cell carcinoma patients are significantly reduced compared with the cancer tissues, and the difference between groups has statistical significance (P < 0.001).

TABLE 1 LncRNA expression statistics

Example 2LncRNA overexpression

1. Cell culture

Human laryngeal squamous carcinoma cell line Hep2, cultured in RPMI1640 medium containing 10% fetal calf serum and 1% P/S at 37 deg.C and 5% CO₂And culturing in an incubator with relative humidity of 90%. The liquid is changed for 1 time in 2-3 days, the cells grow well and grow in a monolayer adherent manner. Passage was routinely digested with 0.25% EDTA-containing trypsin.

2. Transfection

1) Treatment of cells prior to transfection

One day before transfection, 3-5 × 10 are planted on 6-hole culture plates⁵And (3) culturing each cell/hole in an antibiotic-free culture medium for one day, wherein the cell density is 30-50% during transfection, and the cell/hole is replaced by a serum-free culture medium before transfection.

2) LncRNA overexpression

The full-length sequence of the long non-coding RNA was found in NCBI and submitted to pcDNA3.1 for construction of an overexpression plasmid by Shanghai Gima. Coli DH5 α competent cells were used for transformation and plasmid extraction; hep2 was transfected with a validated overexpression plasmid.

3) Transfection

a. Adding 1 μ g/μ l of over-expression plasmid or pcDNA3.1 empty vector into 49 μ l of serum-free culture medium, mixing, and incubating at room temperature for 5 min;

b. mu.l of Lipofectamine 2000 was added to 48. mu.l of serum-free medium. Mixing, and incubating at room temperature for 5 min;

c. mixing the above two mixtures (total volume 100 μ l), gently mixing, and incubating at room temperature for 25min to allow complex formation;

d. adding 100 mul of compound and a proper amount of culture medium into each hole of a 6-hole plate, and gently mixing uniformly;

e. and incubating for 48-96 h, and observing the over-expression condition of the gene.

3. Cell transfection efficiency assay

As a result of the target gene expression measurement using the QPCR method described in example 1, as shown in fig. 1, the expression levels of LOC285627, LOC100506860, and LOC388692 in the cells transfected with the empty vector were set to 100%, and the expression levels of LOC285627, LOC100506860, and LOC388692 in the cells transfected with the LncRNA overexpression vector were significantly increased on average (the difference was statistical,. x P < 0.05;. P <0.01), and the next experiment was performed.

Example 3 Effect of LncRNA expression on laryngeal squamous carcinoma cell proliferation

The WST-1 method detects the influence of LncRNA expression on the proliferation of laryngeal squamous cell carcinoma cells.

The following day of cell transfection, cell density was adjusted to 1 × 10²Mu.l, seeded in 96-well plates (100. mu.l/well) with 3 duplicate wells per set. 37 ℃ and 5% CO₂After 72 hours of culture, 10. mu.l of WST-1 was added and incubated for 2 hours, and then the optical density (D) of the cells was measured at a wavelength of 450nm in a microplate reader.

Results as shown in table 2, the differences between groups were statistically significant (P < 0.05). Shows that the expression of LOC285627, LOC100506860 and LOC388692 is promoted to inhibit the proliferation of laryngeal squamous cell carcinoma cells.

TABLE 2 Callioma cell proliferation statistics

	OD450	SD	T-test
				Empty vector	1.302	0.142
LOC285627 overexpression	0.931	0.009	0.041
				LOC100506860 overexpression	0.795	0.058	0.034
LOC388692 overexpression	0.668	0.039	0.015

Example 4 Effect of LncRNA expression on laryngeal squamous carcinoma cell migration

The following day of cell transfection, the treated cells were collected and counted at 1 × 10⁵The individual cells were resuspended in 100. mu.l serum-free medium and added to the upper chamber of a Transwell cell culture plate, and 600. mu.l complete medium was added to the lower chamber. At 37 ℃ 5% CO₂After 48h incubation, the chamber was removed, the cells in the upper chamber were gently wiped off with a cotton swab, fixed with 4% paraformaldehyde for 20min, washed 1 time with PBS, stained with crystal violet for 10min, washed 1 time with PBS, and the total number of cells in 5 different fields, upper, lower, left, right, and middle, was counted under an optical microscope (× 200), averaged and repeated 3 timesAnd (5) carrying out experiments.

Results as shown in table 3, the differences between groups were statistically significant (P < 0.05). The expression of LOC285627, LOC100506860 and LOC388692 is promoted, and then the cell migration of laryngeal squamous cell carcinoma can be inhibited.

TABLE 3 statistics of laryngeal squamous cell carcinoma cell migration

	Number of cells	SD	T-test
				Empty vector	139.8	15.324
LOC285627 overexpression	115	9.271	0.049
				LOC100506860 overexpression	94.2	5.481	0.029
LOC388692 overexpression	74.4	7.233	0.005

The above description of the embodiments is only intended to illustrate the method of the invention and its core idea. It should be noted that, for those skilled in the art, without departing from the principle of the present invention, several improvements and modifications can be made to the present invention, and these improvements and modifications will also fall into the protection scope of the claims of the present invention.

Sequence listing

<110> Beijing, the deep biometric information technology GmbH

New application of <120> LOC285627

<160>8

<170>SIPOSequenceListing 1.0

<210>1

<211>18

<212>DNA

<213> Artificial Sequence (Artificial Sequence)

<400>1

aatcaagtgg ctaagtca 18

<210>2

<211>21

<212>DNA

<213> Artificial Sequence (Artificial Sequence)

<400>2

ggattcatct cattaactct g 21

<210>3

<211>18

<212>DNA

<213> Artificial Sequence (Artificial Sequence)

<400>3

ccactgagaa gtcctaag 18

<210>4

<211>19

<212>DNA

<213> Artificial Sequence (Artificial Sequence)

<400>4

tgatgatgaa actggctta 19

<210>5

<211>18

<212>DNA

<213> Artificial Sequence (Artificial Sequence)

<400>5

ctcacagtcg gaccttag 18

<210>6

<211>18

<212>DNA

<213> Artificial Sequence (Artificial Sequence)

<400>6

acagaacgga agacaaga 18

<210>7

<211>21

<212>DNA

<213> Artificial Sequence (Artificial Sequence)

<400>7

ctctggtaaa gtggatattg t 21

<210>8

<211>20

<212>DNA

<213> Artificial Sequence (Artificial Sequence)

<400>8

ggtggaatca tattggaaca 20

Claims

1. The application of the reagent for detecting the expression of the long-chain non-coding RNA in the preparation of products for diagnosing laryngeal squamous cell carcinoma; the long non-coding RNA is LOC 285627.

2. The use according to claim 1, wherein the agent comprises: the reagent for detecting the expression level of LOC285627 by reverse transcription PCR, real-time quantitative PCR, immunodetection, in-situ hybridization or chip technology.

3. Use according to claim 2, wherein the agent comprises a nucleic acid which binds to LOC285627 or a DNA sequence thereof.

4. Use according to claim 3, characterized in that said nucleic acid comprises a primer and/or a probe directed against LOC 285627.

5. The use according to claim 4, wherein the primer has the sequence shown in SEQ ID No. 1-2.

6. A product for diagnosing laryngeal squamous carcinoma, wherein the product comprises the agent of any one of claims 1 to 5.

Preferably, the product may be a chip, a kit, a strip, or a high throughput sequencing platform.

More preferably, the kit further comprises instructions for use which recite the following diagnostic criteria: and (3) comparing with a normal control, if the LOC285627 expression in the laryngeal tissues of the suspected canceration of the subject is remarkably reduced, judging that the subject has laryngeal squamous carcinoma or judging that the risk of the subject having laryngeal squamous carcinoma is high.

Use of LOC285627 in the screening of a candidate drug for the prevention or treatment of laryngeal squamous cell carcinoma.

Application of LOC285627 in preparation of medicines for preventing or treating laryngeal squamous cell carcinoma.

9. The use according to claim 8, wherein the medicament comprises a substance that promotes the expression of LOC 285627; preferably, the substance promoting the expression of LOC285627 comprises an overexpression vector containing LOC285627, a host cell comprising the overexpression vector.

10. The pharmaceutical composition for treating laryngeal squamous carcinoma is characterized by comprising a substance for promoting LOC285627 expression, and/or other medicines compatible with the substance and a pharmaceutically acceptable carrier.