CN113652487B

CN113652487B - Application of human C11orf86mRNA in evaluating lung adenocarcinoma patient prognosis and detection kit

Info

Publication number: CN113652487B
Application number: CN202111072265.9A
Authority: CN
Inventors: 赵小刚; 李培超; 田忠献; 李令冰; 杨凌霄
Original assignee: Second Hospital of Shandong University
Current assignee: Second Hospital of Shandong University
Priority date: 2021-09-14
Filing date: 2021-09-14
Publication date: 2022-04-05
Anticipated expiration: 2041-09-14
Also published as: CN113652487A

Abstract

The invention relates to application of human C11orf86mRNA in evaluating lung adenocarcinoma patient prognosis and a detection kit. The invention firstly proves that the abnormal high expression of C11orf86mRNA can promote the proliferation and migration of lung adenocarcinoma cells, and the up-regulated expression level of the C11orf86mRNA is obviously related to the shortening of the overall survival period of lung adenocarcinoma patients. Therefore, the C11orf86mRNA is taken as a biomarker of the lung adenocarcinoma, and the application of the C11orf86mRNA in the lung adenocarcinoma is provided, especially the application in the preparation of a lung adenocarcinoma prognosis evaluation product.

Description

Application of human C11orf86mRNA in evaluating lung adenocarcinoma patient prognosis and detection kit

Technical Field

The invention relates to application of human C11orf86mRNA in evaluating lung adenocarcinoma patient prognosis and a detection kit, and belongs to the technical field of biomedicine.

Background

Lung cancer has become the most common human malignancy, with over 200 million new cases diagnosed each year worldwide. Non-small cell lung cancer is the main pathological type of lung cancer, accounting for about 85% of diagnosed cases of lung cancer, and comprises lung adenocarcinoma, lung squamous carcinoma and large cell carcinoma, wherein lung adenocarcinoma accounts for about 40% of primary lung cancer, and is the most common non-small cell lung cancer. In recent years, despite some progress in early diagnosis and individualized treatment of lung adenocarcinoma, the prognosis of lung cancer patients is still not ideal, and the five-year survival rate is less than 20%. Therefore, the discovery of a new regulatory gene closely related to the occurrence and development of the lung adenocarcinoma is of great significance for further exploring the complex tumor biological characteristics of the lung adenocarcinoma and promoting the development of the clinical diagnosis and treatment strategy of the lung adenocarcinoma.

Polymerase Chain Reaction (PCR) is a technique for specifically amplifying a target DNA fragment in vitro by using the base complementary pairing principle. The reverse transcription-real-time fluorescence quantitative PCR comprises two links of reverse transcription synthesis of a first cDNA chain and real-time fluorescence quantitative PCR. Reverse transcription is performed by using mRNA as a template and synthesizing a cDNA strand complementary to the mRNA as the template by reverse transcriptase. The real-time fluorescent quantitative PCR takes cDNA as a template, a target segment between an upstream primer and a downstream primer is specifically synthesized under the action of DNA polymerase, a fluorescent signal embedded in a DNA double strand can carry out real-time detection on the PCR process, and the target mRNA can be quantified according to a Ct value. Designing a specific primer sequence capable of identifying a target fragment is an important link for ensuring the accurate quantification of target mRNA.

Disclosure of Invention

Aiming at the defects of the prior art, the invention provides an application of human C11orf86mRNA in evaluating the prognosis of a lung adenocarcinoma patient and a detection kit.

The technical scheme of the invention is as follows:

application of human C11orf86mRNA in preparing lung adenocarcinoma prognosis evaluation products.

Preferably according to the invention, the C11orf86mRNA in said use is a biomarker for prognostic assessment of lung adenocarcinoma.

Preferably, according to the invention, the nucleotide sequence of the C11orf86mRNA is shown in SEQ NO. 4.

Preferably, according to the invention, the product is used for assessing and predicting the overall survival of patients with lung adenocarcinoma.

Preferably, according to the present invention, the lung adenocarcinoma prognosis evaluation product comprises a substance that specifically recognizes cDNA, which is a reverse transcription product of C11orf86 mRNA.

Further preferably, the substance specifically recognizing the reverse transcription product cDNA of C11orf86mRNA is selected from a primer pair specifically amplifying the reverse transcription product cDNA of C11orf86 mRNA.

Further preferably, the primer pair for specifically amplifying cDNA of the reverse transcription product of the C11orf86mRNA is an upstream primer shown in SEQ ID NO.1 and a downstream primer shown in SEQ ID NO. 2; or an upstream primer shown in SEQ ID NO.1 and a downstream primer shown in SEQ ID NO. 3.

According to a preferred embodiment of the present invention, the test sample of the lung adenocarcinoma prognosis evaluation product is selected from cells or tissues.

The application of a substance specifically recognizing cDNA of a reverse transcription product of C11orf86mRNA in a lung adenocarcinoma prognosis evaluation product.

Preferably according to the present invention, the substance specifically recognizing the reverse transcription product cDNA of C11orf86mRNA is selected from a primer pair specifically amplifying the reverse transcription product cDNA of C11orf86 mRNA.

A prognostic evaluation kit for lung adenocarcinoma, comprising a substance that specifically recognizes cDNA, which is a reverse transcription product of C11orf86 mRNA.

Preferably, the kit further comprises a detection reagent for real-time fluorescent quantitative PCR.

Has the advantages that:

1. the invention proves that the upregulation of C11orf86mRNA expression can promote the proliferation and migration of lung adenocarcinoma cells for the first time, and the upregulated C11orf86mRNA expression level is obviously related to the shortening of the overall survival period of lung adenocarcinoma patients. Therefore, the C11orf86mRNA is used as a biomarker of the lung adenocarcinoma, and the application of the C11orf86mRNA in the lung adenocarcinoma is provided, especially the application in the preparation of a lung adenocarcinoma prognosis evaluation product.

2. The invention designs a primer pair for specifically amplifying cDNA of a reverse transcription product of C11orf86mRNA, and can effectively detect the expression level of C11orf86 mRNA.

Drawings

FIG. 1 is a graph showing the statistical significance of analysis of expression differences in the Student's t test on the calculation of the relative expression level of C11orf86mRNA in 524 lung adenocarcinoma tissues, based on the relative expression level of C11orf86mRNA in 59 adjacent normal lung tissues, analyzed by RNA sequencing data of lung adenocarcinoma in TCGA database according to the present invention.

FIG. 2 is a graph showing the expression level of C11orf86mRNA in 15 pairs of lung adenocarcinoma tissue and adjacent normal lung tissue, with reference to GAPDH and relative expression level of C11orf86mRNA in adjacent normal lung tissue, according to 2^-△△CTThe relative expression level of C11orf86mRNA in cancer tissues was calculated and a statistical representation of the differences between groups was analyzed using the Student's t test.

FIG. 3 is a graph showing that the kit of the present invention detects the expression level of C11orf86mRNA in normal bronchial epithelial cells (BEAS-2B) and 3 lung adenocarcinoma cell lines (A549, H1650, H1975) with GAPDH as an internal reference and the relative expression level of C11orf86mRNA in BEAS-2B cells as a standard according to 2^-△△CTRelative expression levels of C11orf86mRNA were calculated in 3 lung adenocarcinoma cell lines and a statistical significance profile of differences between groups was analyzed using the Student's t test.

FIG. 4 is a line graph (left) and a bar graph (right) showing the statistical significance of differences between groups analyzed using the Student's t test to examine the effect of exogenous C11orf86mRNA expression on proliferation and migration of BEAS-2B cells using the CCK-8 assay and the Transwell assay.

FIG. 5 is a line graph (left) and a bar graph (right) showing the statistical significance of differences between groups analyzed using the Student's t test to examine the effect of siRNA-specific knockdown of C11orf86mRNA expression levels on the proliferation and migration abilities of A549 cells using the CCK-8 assay and the Transwell assay.

FIG. 6 is a graph showing the effect of C11orf86mRNA expression levels on lung adenocarcinoma patient prognosis using the Kaplan-Meier Plotter database and using Log-rank to test whether the difference in prognosis between patients with high and low expression of C11orf86mRNA is statistically significant.

Detailed Description

The technical solution of the present invention is further described below with reference to the experimental examples, but the scope of the present invention is not limited thereto. The reagents and materials used in the examples are, unless otherwise specified, all of which are commonly commercially available products.

The example was approved by the ethical committee of medical science of the second hospital of Shandong university, and all cases received informed consent from the patients.

Human lung adenocarcinoma cell lines A549, H1650, H1975 and normal human bronchial epithelial cell line BEAS-2B were purchased from the cell bank of the culture Collection of the Chinese academy of sciences.

Example 1

The present invention utilizes RNA-seq (Ribo-free) to sequence mRNA in lung adenocarcinoma tissue and samples of adjacent normal lung tissues (7 cases each), and the results show that C11orf86mRNA expression is detected in 4 cases of lung adenocarcinoma tumor tissue, and C11orf86mRNA expression is not detected in 7 cases of adjacent normal lung tissues, and the inventors further analyzed the RNA sequencing data of lung adenocarcinoma patients in TCGA database, and found that the expression level of C11orf86mRNA in lung adenocarcinoma tissue is significantly increased compared with that in adjacent normal tissues, and the specific analysis results are shown in FIG. 1.

The specific implementation process is as follows:

(1) collecting a tissue sample: collecting 7 pairs of lung adenocarcinoma specimens removed by operation, wherein the tumor tissues are all taken from the central non-necrotic part, the adjacent normal tissues are taken from the area which is more than 5cm away from the tumor edge, and the specimens are quickly frozen by liquid nitrogen and preserved at-80 ℃. No radiotherapy and chemotherapy was performed on 7 patients before surgery, and the pathological result after surgery was confirmed to be lung adenocarcinoma. The above case collections were approved by the ethical committee of medical science of the second hospital of Shandong university and were informed consent from the patients;

(2) RNA sequencing: cracking the frozen and stored tissue of the liquid nitrogen by using a Trizol reagent, extracting RNA by using a phenol-chloroform-isopropanol method, detecting the concentration and the purity of the extracted RNA by using a NanoDrop2000 method, evaluating the integrity of the extracted RNA by agarose gel electrophoresis, taking an RNA sample with RIN >7 for library construction and sequencing, and sequencing by using an RNA-seq (Ribo-free) technology by Beijing Nuo Poa genesis bioinformatics science and technology Limited;

(3) bioinformatics analysis: analyzing the original data obtained by sequencing, sequentially performing quality evaluation, eliminating low-quality sequences, comparing with reference genome, performing differential expression analysis according to (Log)₂ Fold change>2，p value<0.05) or (Log)₂ Fold change<-2，p value<0.05) standard screening for differentially expressed mRNA;

the results showed that no C11orf86mRNA expression was detected in 7 adjacent normal lung tissues, but C11orf86mRNA expression was detected in 4 lung adenocarcinoma tumor tissues;

in view of the above findings, the present inventors continued to analyze the data of RNA sequencing of lung adenocarcinoma in the TCGA database, wherein the results of bioinformatics analysis of 524 cases of lung adenocarcinoma tissue and 59 cases of adjacent normal lung tissue showed that the expression level of C11orf86mRNA was significantly increased in lung adenocarcinoma tissue compared to adjacent normal lung tissue.

Example 2

Collecting 15 samples of the lung adenocarcinoma tissue excised by the operation and the adjacent normal tissue, extracting total RNA, and detecting the expression level of C11orf86mRNA in the sample by using the primer sequence of the cDNA of the reverse transcription product of the C11orf86mRNA specifically identified by the invention to perform real-time fluorescent quantitative PCR, wherein the result shows that the expression level of the C11orf86mRNA in the lung adenocarcinoma tissue is obviously increased compared with the adjacent normal tissue, and the specific analysis result is shown in figure 2.

The specific implementation process is as follows:

(1) collecting a tissue sample: collecting 15 pairs of lung adenocarcinoma specimens, collecting tumor tissue from central non-necrotic region, collecting the region more than 5cm away from the tumor edge adjacent to normal lung tissue, and quick freezing with liquid nitrogen and storing at-80 deg.C. The pathological results of 15 patients after operation proved to be lung adenocarcinoma, and the patients did not receive radiotherapy and chemotherapy before the operation. The above case collections were approved by the ethical committee of medical science of the second hospital of Shandong university and were informed consent from the patients.

(2) Extracting total RNA of tissues: total RNA was extracted from adenocarcinoma lung tissue and adjacent normal lung tissue using RNA-Quick Purification Kit (RN 001), exactly as described in the specification, and the integrity of the extracted RNA was checked by agarose gel electrophoresis and the concentration and purity of the RNA was checked using NanoDrop 2000.

(3) RT-qPCR: removing genomic DNA by using an lnRcute lncRNA cDNA first strand synthesis kit (TIANGEN, KR202), and performing reverse transcription to synthesize cDNA, specifically according to the steps of the specification, using Power SYBR^TMGreen PCR premix (Thermo Fisher Scientific, 4367659) and specific primers of the present invention shown in SEQ No.1, SEQ No.2 and SEQ No.3 were subjected to fluorescent quantitative PCR using GAPDH as an internal control and 2^-ΔΔCTThe method calculates the relative expression level of C11orf86mRNA, and the result shows that the expression level of C11orf86mRNA is obviously increased in lung adenocarcinoma tissues compared with the adjacent normal lung tissues.

Wherein, the nucleotide sequence of the primer pair is as follows:

upstream primer # 1: 5'-CACAGAGCAGCTGATCCAA-3' (SEQ ID NO.1)

Downstream primer # 1: 5'-GCTCTCCCACCTTCTTCTTAC-3' (SEQ ID NO.2)

Downstream primer # 2: 5'-CACCTTCTTCTTACCTGCTGT-3' (SEQ ID NO.3)

Genomic DNA removal system: RNA (250 ng/. mu.L) 2. mu.L, 5 XgDNA Buffer 2. mu.L, RNase-Free ddH₂O6 μ L; reaction conditions are as follows: incubate at 42 ℃ for 3 minutes and place on ice until ready for use.

Reverse transcription PCR System: 10 XlnR RT Buffer 2. mu.L, lnR RT Enzyme Mix 1. mu.L, lnR-RT Primer Mix 2. mu.L, RNase-Free ddH₂O5 mu L, genome DNA removal system product 10 mu L; reaction conditions are as follows: incubate at 42 ℃ for 15 minutes, incubate at 95 ℃ for 3 minutes, and store at 4 ℃ for future use.

Real-time fluorescent quantitative PCR system: reverse transcription System product 1. mu.L, Power SYBR ^TM5. mu.L of Green PCR premix, 1. mu.L of primer diluent (1. mu.M), RNase-Free ddH₂O3. mu.L. Reaction conditions are as follows: incubating at 95 ℃ for 10 minutes; incubating at 95 ℃ for 15 seconds and 60 ℃ for 30 seconds for 40 cycles; incubate at 95 ℃ for 15 seconds, 60 ℃ for 60 seconds, and 95 ℃ for 15 seconds.

The above reagents are all from the first strand synthesis kit (TIANGEN, KR202) and Power SYBR of lnRCUTE lncRNA cDNA^TMGreen PCR premix kit (Thermo Fisher Scientific, 4367659).

Example 3

The expression levels of C11orf86mRNA in normal bronchial epithelial cells (BEAS-2B) and 3 lung adenocarcinoma cell lines (A549, H1650 and H1975) were respectively detected by using the primer sequences of the cDNA specifically recognizing the reverse transcription product of C11orf86mRNA shown in SEQ NO.1, SEQ NO.2 and SEQ NO.3 of the present invention, and the results showed that the expression levels of C11orf86mRNA in A549, H1650 and H1975 cells were significantly increased compared to the BEAS-2B cells, and the specific analysis results are shown in FIG. 3.

The specific implementation process is as follows:

(1) extracting total RNA of cells: total RNA was extracted from normal human airway epithelial cells (BEAS-2B) and 3 lung adenocarcinoma cells (A549, H1650, H1975) using RNA-Quick Purification Kit (Shanghai Yiniao, RN001), integrity of the extracted RNA was checked by agarose gel electrophoresis, and concentration and purity of the RNA were checked using NanoDrop 2000.

(2) RT-qPCR: using a first strand synthesis kit (TIANGEN, KR202) of lnRCUTE lncRNA cDNA, firstly removing genome DNA, then carrying out reverse transcription to synthesize cDNA, specifically according to the steps of the specification, using specific primers shown as SEQ NO.1, SEQ NO.2 and SEQ NO.3 and Power SYBR in the invention^TMGreen PCR premix (Thermo Fisher Scientific, 4367659) was subjected to quantitative fluorescence PCR using GAPDH as an internal reference and 2^-ΔΔCTThe method calculates the relative expression level of C11orf86mRNA, and the result shows that the expression level of C11orf86mRNA in 3 lung adenocarcinoma cell lines is significantly up-regulated compared with BEAS-2B cells.

The PCR system was the same as in example 2.

Example 4

The method comprises the steps of constructing a BEAS-2B cell strain with stably expressed exogenous C11orf86mRNA by using lentivirus, detecting the influence of exogenous C11orf86mRNA expression on the proliferation and migration of the BEAS-2B cell by using a CCK-8 experiment and a Transwell experiment, and showing that the proliferation and migration capacity of the BEAS-2B cell can be remarkably enhanced by increasing the expression of the C11orf86mRNA, wherein the specific result is shown in figure 4.

The specific implementation process is as follows:

(1) construction of BEAS-2B cell line stably expressing exogenous C11orf86 mRNA: the C11orf86 plasmid and the empty vector plasmid were synthesized by Biotech, Inc., of Okagaceae, Beijing. The PMD2G, PSPAX2 and C11orf86 plasmids or the empty vector plasmids were transfected into 293T cells using Lipo293 transfection reagent (Beyotime, C0521), the medium was collected after 48 hours, filtered using a 0.45 μm filter, 5 × PEG8000 solution was added to the filtrate, centrifuged after standing overnight at 4 ℃, the pellet was retained, and the lentiviral pellet was resuspended using serum-free DMEM medium. The BEAS-2B cell strain (BEAS-2B-C11orf86) and the stable expression empty Vector cell strain (BEAS-2B-Vector) which are stably expressed by exogenous C11orf86mRNA can be obtained by infecting the BEAS-2B cell with the obtained lentivirus and screening drug-resistant cell strains by using a culture medium containing puromycin (1 mu g/mL).

(2) Verification of the mRNA table of BEAS-2B-C11orf86 and C11orf86 in BEAS-2B-Vector cellsReaching the level: the total RNA of the cells was extracted using RNA-Quick Purification Kit (RN 001, Shanghai) and cDNA was synthesized by reverse transcription using lnRCute lncRNA cDNA first strand synthesis Kit (TIANGEN, KR202), and Power SYBR was used^TMGreen PCR premix (Thermo Fisher Scientific, 4367659) and the primer sequences of the present invention shown in SEQ NO.1, SEQ NO.2 and SEQ NO.3 that specifically recognize cDNA of mRNA reverse transcription product of C11orf86 were subjected to fluorescent quantitative PCR using GAPDH as an internal reference and 2^-ΔΔCTThe method calculates the relative expression level of C11orf86 mRNA.

(3) CCK-8 experiment: when the cell growth density reached about 80%, cells were digested and collected using trypsin, the cell pellet was retained by centrifugation, cells were resuspended by adding fresh medium, the cell count and cell concentration was adjusted to 15,000/mL medium, 100 μ L of the above cell suspension was pipetted into each culture well (96-well plate), 6 replicate wells were set for each cell, 10 μ L of CCK-8 reagent (TargetMol, C0005) was added to each well every 24 hours, the 96-well plate was incubated at 37 ℃ for 1 hour, and the absorbance value at 450nm was measured and recorded using a microplate reader.

(4) Transwell experiment: when the cell growth density reached about 80%, cells were digested and collected using trypsin, a cell suspension was prepared using serum-free DMEM medium, the cells were counted and adjusted to a cell concentration of 100,000 per mL of the medium, 600 μ L of complete medium was added to each well in a 24-well plate, and a Transwell cell (Corning, 3422) was placed in the 24-well plate to avoid generation of air bubbles, 200 μ L of the above cell suspension was aspirated and added to the cell, the cells in the cell were placed in a 37 ℃ incubator for 48 hours, the medium was discarded, the cells were perforated on the outer side of the cell using 4% paraformaldehyde fixed at room temperature for 15 minutes, washed once with PBS, the cells were treated at room temperature with methanol for 25 minutes, stained with 0.1% crystal violet for 30 minutes, the cells that were not perforated on the inner side of the cell were carefully scraped off with a cotton swab, the perforated cells were observed using an inverted microscope and counted.

Example 5

The siRNA technology is used for specifically knocking down the expression level of C11orf86mRNA in A549 cells, CCK-8 experiments and Transwell experiments are used for detecting the influence of the expression level down-regulation of C11orf86mRNA on the proliferation and migration of the A549 cells, and the results show that the C11orf86mRNA expression down-regulation obviously inhibits the proliferation and migration capacity of the A549 cells, and the specific results are shown in FIG. 5.

The specific implementation process is as follows:

(1) specific knock-down of a549 cells C11orf86mRNA expression levels using siRNA technology: the RNAi Designer website was used to design C11orf86 sirnas that specifically interfere with C11orf86mRNA expression and were synthesized by the biotechnology company, kyoto.

Wherein the C11orf86siRNA targeting nucleotide sequence is: 5'-atgattctatgaaggtttaga-3', respectively;

(2) c11orf86siRNA or negative control (si-NC) were transfected into a549 cells and tested for knockdown efficiency: c11orf86siRNA or si-NC was transfected into A549 cells using Lipofectamine RNAImax Reagent (Life technologies, 13778-^TMGreen PCR premix (Thermo Fisher Scientific, 4367659) and the primer sequences of the present invention shown in SEQ NO.1, SEQ NO.2 and SEQ NO.3 that specifically recognize cDNA of mRNA reverse transcription product of C11orf86 were subjected to fluorescent quantitative PCR using GAPDH as an internal reference and 2^-ΔΔCTThe method calculates the relative expression level of C11orf 86.

(3) CCK-8 experiment: c11orf86siRNA or si-NC was transfected into A549 cells using Lipofectamine RNAImax Reagent (Life technologies, 13778-.

(4) Transwell experiment: transfection of C11orf86siRNA or si-NC into A549 cells using Lipofectamine RNAImax Reagent (Life technologies, 13778-.

Example 6

The Kaplan-Meier Plotter database was used to analyze the effect of C11orf86mRNA expression levels on lung adenocarcinoma patient prognosis, and the results showed that: compared with the prognosis of lung adenocarcinoma patients with low expression of C11orf86mRNA, the overall survival of lung adenocarcinoma patients with high expression of C11orf86mRNA was significantly shortened, and the specific results are shown in FIG. 6.

The specific implementation process is as follows:

downloading prognosis information and relative expression level of C11orf86mRNA of lung adenocarcinoma patients in a Kaplan-Meier Plotter database, wherein 672 cases of lung adenocarcinoma patients are total, and analyzing the prognosis difference of C11orf86mRNA high-expression and low-expression patients by applying a Kaplan-Meier method, wherein the C11orf86mRNA is highly expressed for 242 cases, and the median survival time is 79.87 months; 430 cases of C11orf86mRNA were underexpressed, median survival 119.87 months, and statistical significance of survival differences among groups was analyzed using the Log-rank test, showing a P value of less than 0.0001.

The above-mentioned embodiments are merely preferred embodiments of the present invention, and are not intended to limit the present invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

SEQUENCE LISTING

<110> secondary Hospital of Shandong university

Application of <120> human C11orf86mRNA in evaluating lung adenocarcinoma patient prognosis and detection kit

<160> 4

<170> PatentIn version 3.5

<210> 1

<211> 19

<212> DNA

<213> Artificial sequence

<400> 1

cacagagcag ctgatccaa 19

<210> 2

<211> 21

<212> DNA

<213> Artificial sequence

<400> 2

gctctcccac cttcttctta c 21

<210> 3

<211> 21

<212> DNA

<213> Artificial sequence

<400> 3

caccttcttc ttacctgctg t 21

<210> 4

<211> 1168

<212> RNA

<213> Homo sapiens

<400> 4

agagggccag ugucuugcug aggggccgga gcaguccugu gccugcagcc uccggagcca 60

ugggaacagg gcugcgaagu caguccuugc gagagccccg acccuccuau ggaaagcugc 120

aggagcccug ggggaggccc caggagggcc aacuccgcag ggcgcuaagc cucagacagg 180

ggcaagagaa guccaggucc cagggccucg agagaggcac agaagggcca gaugccacug 240

cccaggagcg ggugccgggg agccuggggg acacagagca gcugauccaa gcccagcgaa 300

gaggcagccg gugguggcug aggcgguacc aacagcaggu aagaagaagg ugggagagcu 360

uugucgccau cuuccccagc gugacucuga gucagccggc cuccccguag cccacacugg 420

gcaccaucag cuaaagaugc uggaagccau cguggccccc uugcugugcc uggaccagcc 480

caccgugucu gcugaccuac cucuucacag cucucuggac uuuggcuggg uggcccuagg 540

aggcuucugc caccagcuca cuggagucgc cacguggcug ccacucuuag cucuggcccu 600

uccacuccag cacuagcucu cuuugaagau cccagcaggg ucaaaaagaa gggggcucag 660

cugucugccc ucugggcuug gguaggggcc uuggacuaug auucuaugaa gguuuagaag 720

aagccugccc ggaauggggg cuguggacgc uccugcccca cacgaccagc ugucaacuuc 780

caggacggag cuugaugaag ccagacccau ggccgagagg cucacggacg uugccugguc 840

acugggccau gacccaacug cccuucuugu caguugcuga uggugggcca gggcgcaggu 900

cucccuccca gauggugagc ccucuggggc aaggaccaca gcuggguggc ccuccuggcu 960

gagcacucug ugcugaaaac cuuugaaccu cacggugucc ugaugaagga agcagaggaa 1020

gucuuccucc cugugugaca gagggggaaa cugaggccca gaguggggaa gggauuucuu 1080

gguuuugaca ucuccgaccc caccccaccc cauccagugu ccagcacugg agucgaacac 1140

aguaauaaag augcugagaa aaagucaa 1168

Claims

1. Application of human C11orf86mRNA in preparing lung adenocarcinoma prognosis evaluation products.

2. The use of claim 1, wherein the C11orf86mRNA is a biomarker for prognostic assessment of lung adenocarcinoma.

3. The use of claim 1, wherein the nucleotide sequence of C11orf86mRNA is set forth in SEQ No. 4.

4. The use of claim 1, wherein the product is used to assess and predict overall survival in a patient with lung adenocarcinoma.

5. The use according to claim 1, wherein the lung adenocarcinoma prognosis evaluation product comprises a primer pair specifically amplifying C11orf86mRNA reverse transcription product cDNA; the primer pair for specifically amplifying cDNA of a reverse transcription product of C11orf86mRNA is an upstream primer shown in SEQ ID NO.1 and a downstream primer shown in SEQ ID NO. 2; or an upstream primer shown in SEQ ID NO.1 and a downstream primer shown in SEQ ID NO. 3.

6. The use of claim 1, wherein the test sample of a product for the prognosis evaluation of lung adenocarcinoma is selected from the group consisting of cells and tissues.