CN109880906A - Oral cavity squamous carcinoma diagnosis target gene and its application - Google Patents

Abstract

The invention discloses oral cavity squamous carcinoma diagnosis target gene and its applications.The present invention passes through screening expression gene relevant to Tumor Differentiation grade, determine the expression of gene in tumor tissues in each differentiation grade so that it is determined that tumour differentiation grade, thus accurately, the formulation and curative effect monitoring of objective prediction tumor prognosis, guiding treatment scheme.

Description

Oral cavity squamous carcinoma diagnosis target gene and its application

Technical field

The invention belongs to biomedicine fields, are related to oral cavity squamous carcinoma diagnosis target gene and its application, are specifically related to Gene is IQCD.

Background technique

Oral squamous cell carcinoma (oral squamous cell carcinoma, OSCC) is a kind of very disruptive and cause The malignant tumour of life occupies significant proportion in all head-neck malignant tumors, has become a big killer of human health. Oral squamous cell carcinoma often seriously affects the life quality of patient and most prognosis malas because of its special anatomical position.People Daily bad habit, such as smoking and drinking is the risk factor of cancer metastasis, and the transfer of tumour is cancer patient face The serious problems faced.According to statistics, the cancer related mortality rate more than 90% is due to transfer.Work as in oral squamous cell carcinoma In, there is lymphatic metastasis in the patient for having more than 50%, and 5 years survival rates of these patients are remarkably decreased.

The predilection site of oral squamous cell carcinoma is followed successively by tongue, gum, cheek, palate, lip, mouth bottom.Squamous cell carcinoma is with angling It is pathological characteristics that pearl, which forms and occur intercellular bridge,.Tumour comes from surface Oral mucosa keratinocyte, tumprigenicity epithelial mass or epithelium Island infiltration lower section connective tissue, cell characteristic can behave as oxyplasm abundant, and karyon, karyoplasmic ratio are big, and degree is different The pleomorphism of cell and karyon, the keratin pearl and single celled angling that scaly epithelium is formed are visible.The tissue of squamous cell carcinoma What histological grading was applied at present is Broder staging.I grades (differentiated): on histology, cytologic characteristic and Normal oral mucosa It is skin-deep seemingly.Basal cell and there is the squamous cell ratio of intercellular bridge to differ, angling is obvious, and mitosis figures are few, is not true to type point It splits and apocyte is rare, cell and karyon pleomorphism are unobvious；II grades of (middle differentiation) histological appearances are between I grades and III level Between, compared with I grades, angling is few, and the pleomorphism of cell and karyon is more apparent, and nuclear fission increases, accidental exception nuclear fission, cell Between bridge it is unobvious；III level (low differentiation): histology, Cytological Characteristic are only slightly similar to Normal oral mucosa epithelium.Angling and Intercellular bridge is rare, and mitosis figures are common, common atypical karyokinesis, and cell and karyon pleomorphism are obvious, and apocyte is more See.

Under the drive of molecular biology fast development, traditional tumour parting, classification and clinical value and meaning by stages Also different degrees of variation is generated therewith, and scholars are dedicated to studying the pathogenesis of oral squamous cell carcinoma further.At this stage The focus mostly on biological characteristics of the oncogene and tumor suppressor gene that have been found that in analysis of research hotspot seek peace in terms of function, discovery is new Tumor markers provide new thinking for the research of oral squamous cell carcinoma pathogenesis, it is early to find oral squamous cell carcinoma Phase diagnostic flag and effective treatment method provide scientific basis, so that the individualized treatment for tumour provides more accurate point Sub- biological information, the formulation and curative effect monitoring for instructing individualized treatment scheme.

Summary of the invention

In order to make up for the deficiencies of the prior art, the purpose of the present invention is to provide a kind of development process phases with oral squamous cell carcinomas The biomarker of pass uses the marker, it can be estimated that the cancer of Patients With Oral Squamous Cell Carcinoma is classified, to realize oral squamous cell carcinomas Precisely treatment.

To achieve the goals above, the present invention adopts the following technical scheme:

The first aspect of the present invention provides product of the reagent in preparation assessment oral squamous cell carcinomas differentiation grade of detection IQCD In application.

Further, the differentiation grade of oral squamous cell carcinomas is selected from the mouth of the oral squamous cell carcinomas of good differentiation (differentiated), medium differentiation The oral squamous cell carcinomas of chamber squamous carcinoma or bad differentiation (low differentiation).

Further, the reagent includes by RT-PCR, real-time quantitative PCR, immune detection, in situ hybridization or chip detection The reagent of IQCD expression in sample.Wherein, the sample for detecting IQCD include cell, tissue, internal organs, body fluid (blood, Lymph etc.), digestive juice, expectoration, alveole bronchus cleaning solution, urine, excrement etc..Preferably, the sample is tissue, blood. In a specific embodiment of the invention, the sample is tissue.

Further, the reagent is selected from: oligonucleotide probe, the specific amplification IQCD base of specific recognition IQCD gene The primer of cause or the antibody or ligand for specifically binding IQCD albumen.

Further, the primer sequence of specific amplification IQCD gene is as shown in NO.1~2 SEQ ID.

Further, expression of the IQCD in the oral squamous cell carcinomas of different differentiation grades is different, and wherein differentiation degree is lower, The expression of IQCD is higher.

The second aspect of the present invention provides a kind of kit for assessing oral squamous cell carcinomas development process, and the kit includes Carry out DNA microarray, oligonucleotide microarray, protein arrays, RNA blotting, RNase protection test, immunoblotting and DNA chip, oligonucleotide chip necessary to reverse transcriptional PCR, protein chip, probe or primer, to detect IQCD Expression.

Further, the kit by the method for DNA microarray, reverse transcriptional PCR or protein arrays into Row detection.

It further, include the primer of specific amplification IQCD by the reagent that reverse transcriptional PCR is detected, Preferably, the sequence of the primer is as shown in NO.1~2 SEQ ID.

Further, expression of the IQCD in the oral squamous cell carcinomas of different development processes is different, wherein differentiation degree it is high, Expression is relatively low in the low oral squamous cell carcinomas of grade malignancy, the table in the oral squamous cell carcinomas that differentiation degree is low, grade malignancy is high It is relatively high up to level.

The third aspect of the present invention provides IQCD answering in the computation model of building assessment oral squamous cell carcinomas differentiation grade With.

The fourth aspect of the present invention provides application of the IQCD in the pharmaceutical composition of preparation treatment oral squamous cell carcinomas.

The fifth aspect of the present invention provides application of the IQCD in the drug candidate of screening treatment oral squamous cell carcinomas.

Kit of the present invention can be used for detecting multiple genes including IQCD gene (for example, with oral cavity squama The relevant multiple genes of cancer development process) expression.The protein chip or protein immunization detection kit can be used for examining Survey the expression of multiple protein (such as multiple protein relevant to oral squamous cell carcinomas development process) including IQCD albumen It is horizontal.Multiple markers of oral squamous cell carcinomas degree of danger are detected simultaneously, oral squamous cell carcinomas degree of danger is greatly improved and examines Disconnected accuracy rate.

The advantages of the present invention:

The present invention selects IQCD as molecular marker, and the differentiation of oral squamous cell carcinomas can be judged according to the expression of IQCD Grade, so that doctor is instructed to take different therapeutic strategies, means and measure to the Patients With Oral Squamous Cell Carcinoma in different development processes, And then can be to avoid over-treatment, it can also be insufficient to avoid treatment intensity, realize the individualized treatment of Patients With Oral Squamous Cell Carcinoma.

Detailed description of the invention

Fig. 1 is the expression figure using QPCR detection IQCD in oral squamous cell carcinoma tissues.

Detailed description of the invention

The present invention after extensive and in-depth study, by high-flux sequence and bioinformatic analysis, screening from G1 is to medium differentiation OSCC (G2) and from G2 to bad base for breaking up its in OSCC (G3) transition process and expressing significant changes Cause.With the selected differentiation etc. expressed the gene significantly changed in each differential period and can be correctly predicted tumor tissues Grade.

In the present invention, it would be recognized by those skilled in the art that practicability of the invention is not limited to of the invention The gene expression of any specific variants of marker gene is quantified.The IQCD gene is in current international public nucleic acid data ID in the GeneBank of library is 115811.

IQCD

IQCD covers IQCD and its homologue, mutation and isoform.IQCD covers overall length, unprocessed IQCD, Yi Jiyuan Any type of IQCD processed from cell.The term covers natural generation variant (such as splice variant or the equipotential of IQCD Variant).The term covers such as IQCD gene, mRNA sequence (such as NM_001330452.2 or such as NM_ of people IQCD 001330452.2) and the amino acid sequence of people IQCD such as (NP_001317381.1 or as NP_001317381.1)；And come From any other vertebrate origin, including mammal, such as Primate and rodent (such as mouse and rat) IQCD DNA, mRNA and amino acid sequence.

" polynucleotides " or " nucleic acid " are used interchangeably herein, and refer to the nucleotide polymer of any length, including DNA And RNA.Nucleotide can be deoxyribonucleotide, ribonucleotide, nucleotide or base and/or its class by modification Like object, or any substrate in polymer can be mixed by DNA or RNA polymerase, or by synthetic reaction.In this way, For example, polynucleotides as defined herein include but is not limited to single-stranded and double-stranded DNA, the DNA comprising single stranded zone and double stranded region, Single-stranded and double-stranded RNA, and the RNA comprising single stranded zone and double stranded region, the hybrid molecule comprising DNA and RNA, it can be single-stranded , perhaps more typically double-strand or include single stranded zone and double stranded region.In addition, term " polynucleotides " is for herein When refer to three sequences comprising both RNA or DNA or RNA and DNA.Chain in such area may be from identical molecule or from different point Son.The area may include the entire of one or more molecules, but more typically only include an area of some molecules.Three strands One of molecule of helical region is often oligonucleotides.Term " polynucleotides " clearly includes cDNA.

Polynucleotides may include the nucleotide by modification, such as methylated nucleotide and the like.To nucleotide knot The modification of structure can carry out before or after assembling polymer.Nucleotide sequence can be interrupted by non-nucleotide component.Multicore Thuja acid can be modified further in post synthesis, such as by being conjugated with marker.Other types of modification includes such as " cap ", will One or more naturally occurring nucleotide are substituted with analog, and internucleotide modification such as has neutral connection (such as methyl phosphonate, phosphotriester, phosphoramidate (phosphoamidate), carbamate etc.) and there is electrically charged company The modification of (such as thiophosphate, phosphorodithioate etc.) is connect, such as containing pendency module (pendant moiety) The modification of protein (such as nuclease, toxin, antibody, signal peptide, polylysine etc.), having intercalator, (such as acridine is mended Bone fat element etc.) modification, the modification containing chelating agent (such as metal, radioactive metal, boron, oxidisability metal etc.) contains alkane The modification of agent has the modification through modification connection (such as α anomeric nucleic acid) and the polynucleotides of unmodified form. In addition, any hydroxyl group being typically found in carbohydrate can use such as phosphonyl group, phosphate group replacement is protected with standard Radical protection, or activate to prepare other connection with other nucleotide, or solid or semisolid support can be conjugated to.It can Phosphorylation replaces 5 ' and 3 ' end OH with the organic capping group module of amine or 1-20 carbon atom.Other hydroxyls can also spread out Generate standard protecting group.Analog shape of the polynucleotides containing ribose or deoxyribose carbohydrate as commonly known in the art Formula, including such as 2 '-oxygen-methyl-, 2 '-oxygen-allyl-, 2 '-fluoro- or 2 '-nitrine-ribose, carba sugars, α-end group Isomerized sugar, epimerism sugar such as arabinose, xylose or lyxose, pyranose, furanose, sedoheptulose are acyclic similar Object and abasic nucleoside analog such as methylribonucleotide.One or more di-phosphate esters can be replaced with alternative linking group to connect It connects.All connections not in polynucleotides are all necessarily identical.Polynucleotides can be different types of containing one or more Modification and/or the modification of many places same type described herein.Foregoing description is suitable for all multicore glycosides mentioned in this article Acid, including RNA and DNA.

As used herein, " oligonucleotides " refers generally to short, and single-stranded polynucleotides are less than about in length 250 nucleotide, but it's not necessary.Oligonucleotides can be synthesis.Term " oligonucleotides " and " polynucleotides " are simultaneously It is not mutually exclusive.Description above for polynucleotides is same and is applicable to oligonucleotides completely.

" primer " refers to hybridize with nucleic acid and the polymerization of complementary nucleic acid is allowed (generally to pass through offer as used herein 3 ' free-OH groups) single stranded polynucleotide.

The present invention can use any method known in the art measurement gene expression.Those skilled in the art should manage Solution, the means for measuring gene expression are not importances of the invention.It can be detected on transcribing or translating (i.e. albumen) level The expression of biomarker.These technologies include but is not limited to: nucleic acid sequencing, nucleic acid hybridization, nucleic acid amplification technologies, albumen Immunological technique.

The exemplary, non-limitative example of Nucleic acid sequencing techniques includes but is not limited to chain terminator (Sanger) sequencing and dye Expect terminator sequencing.Those skilled in the art it will be recognized that due to RNA in cell less stable and in an experiment It is more vulnerable to nuclease attack, therefore usually by RNA reverse transcription at DNA before sequencing.

The another exemplary non-limiting example of Nucleic acid sequencing techniques includes that (deep sequencing/high pass measures for next-generation sequencing Sequence), high throughput sequencing technologies are a kind of sequencing technologies in synthesis based on unimolecule cluster, based on proprietary reversible termination chemistry Reaction principle.The random fragment of the DNA of genome is attached to optically transparent glass surface when sequencing, these DNA fragmentations warp After crossing extension and bridge amplification, hundreds of millions of clusters is formed in glass surface, each cluster is the list with thousands of parts of same templates Then molecular cluster utilizes four kinds of special deoxyribonucleotides with fluorophor, skill is sequenced in synthesis by reversible Template DNA to be measured is sequenced in art.

The exemplary, non-limitative example of nucleic acid hybridization technique include but is not limited in situ hybridization (ISH), microarray and Southern or Northern trace.In situ hybridization (ISH) be it is a kind of use label complementary DNA or RNA chain as probe with Position tissue a part or slice (original position) are the spy in entire tissue (full organization embedding ISH) if tissue is sufficiently small The hybridization of anisotropic DNA or RNA sequence.DNA ISH can be used for determining the structure of chromosome.RNA ISH is for measuring and positioning group Knit the mRNA and other transcripts (for example, ncRNA) in slice or full organization embedding.Usually to sample cell and tissue at Reason increases the entrance of probe with fixation in situ target transcript.Probe hybridizes with target sequence at high temperature, then by extra spy Needle is washed off.Use autoradiograph, fluorescence microscopy or immunohistochemistry respectively, in tissue with radiation, fluorescence or antigen The probe of the kilobase marker of label is positioned and is quantified.Two or more can also be used by radioactivity by ISH or other are non- The probe of radioactive label substance markers, to detect two or more transcripts simultaneously.

The present invention simultaneously can expand nucleic acid (for example, ncRNA) before detection or with detection.Nucleic acid amplification technologies Exemplary, non-limitative example include but is not limited to: polymerase chain reaction (PCR), reverse transcriptase polymerase chain reaction (RT- PCR), the amplification (TMA) of transcriptive intermediate, ligase chain reaction (LCR), strand displacement amplification (SDA) and based on nucleic acid sequence It expands (NASBA).Those skilled in the art will be it will be recognized that certain amplification techniques (for example, PCR) needs will before amplification RNA reverse transcription is at DNA (for example, RT-PCR), and other amplification techniques then direct cloning RNA (for example, TMA and NASBA).

The polymerase chain reaction of commonly referred to as PCR uses denaturation, the annealing and primer extend of primer pair and opposite strand Multiple circulations, exponentially increase target nucleic acid sequence copy number；The amplification of the transcriptive intermediate of TMA is (substantial constant Temperature, multiple copies of target nucleic acid sequence are autocatalytically synthesized under conditions of ionic strength and pH, wherein target sequence is more A RNA copy autocatalytically generates other copy；The ligase chain reaction of LCR uses miscellaneous with the adjacent area of target nucleic acid The two groups of complementary DNA oligonucleotides handed over；Other amplification methods include for example: the commonly referred to as expansion based on nucleic acid sequence of NASBA Increase；Use the amplification of rna replicon enzyme (commonly referred to as Q β replicase) amplification probe molecule itself；Amplification method based on transcription； And the sequence amplification of self―sustaining.

Protein immunization technology includes sandwich immunoassay, such as sandwich ELISA, wherein using identifying on biomarker not The detection of the biomarker is carried out with two kinds of antibody of epitope；Radiommunoassay (RIA), direct, indirect or comparison are enzyme-linked Immunosorbent assay (ELISA), enzyme immunoassay (EIA) (EIA), fluorescence immunoassay (FIA), immunoblotting, immuno-precipitation With the immunoassays based on any particle (as used gold particle, Argent grain or latex particle, magnetic-particle or quantum dot).It can example Such as implement immunization in the form of microtiter plate or item.

In the present invention, kit includes the reagent and one or more substances selected from the group below for detecting IQCD: being held Device, operation instructions, positive control, negative control object, buffer, auxiliary agent or solvent.

In kit of the invention can also have kit operation instructions, be described how using kit into Row detection, and how tumor development to be judged using testing result, therapeutic scheme is selected.

Antibody used in the present invention for above-mentioned IQCD protein is specifically covered for example single with broadest use Clonal antibody, polyclonal antibody, the antibody with multi-epitope specificity, multi-specificity antibody and antibody fragment.Such antibody can Be it is chimeric, humanization, people's and synthesis.

In the present invention, term " includes " is for referring to phrase " including but not limited to ", and with phrase " including but not limited to " It may be used interchangeably.

Statistical method

In the present invention, experiment is at least tested using 3 repetitions, and result data is all in a manner of mean+SD Indicate, come using SPSS18.0 statistical software for statistical analysis, difference between the two is using t inspection, it is believed that when P < There is statistical significance when 0.05.

Specific embodiment

The present invention is described in further detail with reference to the accompanying drawings and examples.Following embodiment is merely to illustrate this It invents rather than limits the scope of the invention.Test method without specific conditions in embodiment, usually according to conventional strip Part, or according to the normal condition proposed by manufacturer.

Embodiment 1 screens gene marker relevant to oral squamous cell carcinomas

1, sample collection

59 oral squamous cell carcinoma tissues and cancer beside organism are collected respectively, including the trouble for through histological grade being I grades (G1) Person 15, histological grade is patient 23 of II grades (G2), and histological grade is patient 21 of III level (G3/G4), patient It is preoperative not receive any treatment.Every group takes 4 samples to carry out high-flux sequence analysis, carries out the screening of difference expression gene, And confirmatory experiment is carried out in each group whole sample.

2, the preparation and quality testing of RNA sample

The extraction of total serum IgE, step are carried out using the total RNA from animal tissues extracts kit (catalog number (Cat.No.) DP431) of Tiangeng It is as follows:

1) homogenized

Every 10-20mg tissue plus 300 μ l lysate RL are thoroughly ground tissue with grinding pestle；Add then in homogenate Enter 590 μ l RNase-Free ddH₂O and 10 μ l Proteinase K, 56 DEG C of processing 10-20min after mixing.

2) 12,000rpm is centrifuged 2-5min, and supernatant is taken to perform the following operation.

3) it is slowly added to 0.5 times of supernatant volume dehydrated alcohol, is mixed, obtained solution and precipitating is transferred to adsorption column together In CR3 (adsorption column is placed in collecting pipe), 12,000rpm centrifugation 30s discard the waste liquid in collecting pipe, adsorption column are put back to receipts In collector.

4) 350 μ l protein liquid removal RW1,12,000rpm centrifugation 30s are added into adsorption column CR3 and waste liquid are abandoned, by adsorption column It puts back in collecting pipe.

5) the DNase I working solution of 80 μ l is added to the center adsorption column CR3, is placed at room temperature for 15min.

6) 350 μ l protein liquid removal RW1,12,000rpm centrifugation 30s are added into adsorption column CR3 and waste liquid are abandoned, by adsorption column It puts back in collecting pipe.

7) 500 μ l rinsing liquid RW are added into adsorption column CR3, are stored at room temperature 2min, 12,000rpm centrifugation 30s are abandoned useless Liquid puts back to adsorption column CR3 in collecting pipe.

8) step 7) is repeated.

9) 12,000rpm is centrifuged 2min, outwells waste liquid.Adsorption column CR3 is placed in and is placed at room temperature for several minutes, thoroughly to dry Remaining rinsing liquid in adsorbent material.

10) adsorption column CR3 is transferred in a new RNase-Free centrifuge tube, is vacantly dripped to the intermediate position of adsorbed film Add 30-100 μ l RNase-Free ddH₂O, is placed at room temperature for 2min, and 12,000rpm centrifugation 2min obtain RNA solution.

11) quality testing of RNA

With the integrality (deposition condition: gum concentration 1.2% of agarose gel electrophoresis detection RNA；0.5 × TBE running buffer Liquid；150V, 15min) detection integrality.When 28S rRNA is twice of 18S rRNA, illustrate that the integrality of RNA is preferable.

With the concentration and purity of spectrophotometer detection RNA, OD260/OD280 is read between 1.8 and 2.1, the matter of RNA It measures higher.

3, construction cDNA library

1) rRNA in total serum IgE is removed using the Ribo-Zero kit of Epicentre；

2) building of cDNA library is carried out using Illumina TruseqTM RNA sample Prep Kit, it is specific to grasp Make by specification progress.

4, upper machine sequencing

Qualified library is detected, NaOH denaturation is added at single-stranded, it is anticipated that upper machine data volume, is diluted to certain upper machine Concentration.Library after denaturation dilution is added in FlowCell, hybridizes with the connector on FlowCell, bridge-type is completed on cBot PCR amplification is finally sequenced using Illumina Hiseq x-ten platform.

5, high-throughput transcript profile sequencing data analysis

Bioinformatic analysis is carried out to sequencing result, linear by is carried out to sequencing data using tool R-3.3.3 Linear association test analysis, being divided into each sample according to the quartile of the expression quantity of each gene is 4 Then the correlation in expression quantity section and tumor grade is detected in a expression quantity section.When FDR value < 0.05, it is believed that gene Significant difference expression.

6, result

Significant difference is presented in expression quantity of the IQCD gene in the oral squamous cell carcinoma tissues of different differentiation degrees, with G1 phase Expression than IQCD in, G2 and G3/G4 is significantly raised, and compared with G2, the expression of IQCD is significantly raised in G3/G4, prompts IQCD Can be used as possible marker is the Patients With Oral Squamous Cell Carcinoma for distinguishing different differentiation grades.

The differential expression of 2 QPCR sequence verification IQCD gene of embodiment

1, the QPCR verifying for the sample that IQCD gene differential expression is all collected.

2, RNA extraction step is as described in Example 1.

3、QPCR

According to the gene order design primer of IQCD and GADPH, wherein select common between the different transcripts of IQCD Region carries out design of primers, and primer sequence is as shown in table 1.

1 primer sequence of table

Use Quant one-step method reverse transcription-fluorescence quantitative kit (SYBR Green) kit of TIANGEN company (catalog number (Cat.No.): NG105) carries out PCR reaction, and reaction system and reaction condition is as shown in table 2.In Thermal Cycler PCR amplification is carried out on Real Time System amplification instrument, confirms the amplification curve of Real Time PCR and molten after reaction Solution curve, Δ Δ CT method carry out relative quantification.

2 QPCR reaction system and reaction condition of table

5, result

As a result as shown in Figure 1, significant difference is presented in IQCD in the oral squamous cell carcinoma tissues of different development processes, and break up Degree is lower, and grade malignancy is higher, and the expression of IQCD is higher, and IQCD is prompted to can be used as molecular marker applied to oral cavity squama Cancer breaks up the judgement of grade, the formulation and curative effect monitoring of accurate, objective prediction tumor prognosis, guiding treatment scheme.

The explanation of above-described embodiment is only intended to understand method and its core concept of the invention.It should be pointed out that for this For the those of ordinary skill in field, without departing from the principle of the present invention, several improvement can also be carried out to the present invention And modification, these improvement and modification will also be fallen into the protection scope of the claims in the present invention.

Sequence table

<110>No.2 Hospital, Hebei Medical Univ.

<120>oral cavity squamous carcinoma diagnosis target gene and its application

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ttattgacag cctgatag 18

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atgatttgtt ggttattact a 21

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<211> 21

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aatcccatca ccatcttcca g 21

<210> 4

<211> 19

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gagccccagc cttctccat 19

Claims (10)

1. detecting application of the reagent of IQCD in the product of preparation assessment oral squamous cell carcinomas differentiation grade.

2. application according to claim 1, which is characterized in that the differentiation grade of oral squamous cell carcinomas is selected from the oral cavity well broken up The oral squamous cell carcinomas of squamous carcinoma, the oral squamous cell carcinomas of medium differentiation or bad differentiation.

3. application according to claim 1, which is characterized in that the reagent include by RT-PCR, real-time quantitative PCR, The reagent of IQCD expression in immune detection, in situ hybridization or chip detection sample.

4. application according to claim 3, which is characterized in that the reagent is selected from: the probe of specific recognition IQCD, spy The primer of specific amplification IQCD or the antibody or ligand for specifically binding IQCD.

5. application according to claim 4, which is characterized in that the primer sequence of specific amplification IQCD such as SEQ ID Shown in NO.1~2.

6. application according to claim 1-5, which is characterized in that oral squamous cell carcinomas of the IQCD in different differentiation grades In expression it is different, wherein differentiation degree is lower, and the expression of IQCD is higher.

7. a kind of kit of assessment oral squamous cell carcinomas differentiation grade, which is characterized in that the kit includes to carry out the micro- battle array of DNA Column, oligonucleotide microarray, protein arrays, RNA blotting, RNase protection test, immunoblotting and reverse transcription polymerase DNA chip, oligonucleotide chip necessary to chain reaction, protein chip, probe or primer, to detect the expression of IQCD.

8. kit according to claim 7, which is characterized in that the kit is polymerize by DNA microarray, reverse transcription The method of enzyme chain reaction or protein arrays is detected.

9. kit according to claim 8, which is characterized in that the examination detected by reverse transcriptional PCR Agent includes the primer of specific amplification IQCD.

10. kit according to claim 9, which is characterized in that the sequence of the primer such as institute of SEQ ID NO.1~2 Show.