CN104111332B - Based on the kit of protein marker PAK4 auxiliary diagnosis Patients with Non-small-cell Lung - Google Patents

Abstract

The invention discloses a kit for auxiliary diagnosis of patients with non-small cell lung cancer based on the protein marker PAK4. The present invention claims the application of the product for detecting the protein marker PAK4 in the preparation of a kit for auxiliary diagnosis of patients with non-small cell lung cancer. In the present invention, the inventors discovered the existence of the protein marker PAK4 in the peripheral blood of NSCLC patients for the first time, and found that the concentration of the protein marker PAK4 in the peripheral blood of NSCLC patients was higher than that of healthy people, that is, the protein marker PAK4 has an auxiliary diagnosis Value for NSCLC patients. In the present invention, the inventors found through immunohistochemistry that the expression of the protein marker PAK4 in the tumor tissue of NSCLC patients was significantly higher than that in the normal tissue next to the tumor tissue, and the expression intensity was related to the prognosis and survival time, which can be used as a candidate prognostic factor. The invention has great value for auxiliary diagnosis and prognosis of non-small cell lung cancer.

Description

Kit for auxiliary diagnosis of patients with non-small cell lung cancer based on protein marker PAK4

technical field

The invention relates to a kit for auxiliary diagnosis of patients with non-small cell lung cancer based on the protein marker PAK4.

Background technique

The incidence of cancer worldwide is still rising, especially lung cancer, which has become the number one cause of cancer death. In 2013, it is estimated that there will be 228,190 new cases of lung cancer in the United States. Although it only accounts for 14% of new cancer cases, the number of deaths caused by it accounts for 27% of all cancer deaths. Pathologically, lung cancer can be divided into non-small cell lung cancer (NSCLC, accounting for about 85%) and small cell lung cancer (SCLC, accounting for about 15%).

The main treatment methods for lung cancer include surgery, radiotherapy and chemotherapy. In recent years, although tumor treatment technology has made great progress and targeted therapy has achieved certain results, the 5-year survival rate of lung cancer is still hovering around 15%. Studies suggest that such a poor prognosis is mainly due to the late detection of tumors. According to reports, up to 56% of newly discovered lung cancer cases are accompanied by distant metastasis, and only 15% of the lesions are confined to the primary tumor; while the corresponding 5-year survival rates are 3.7% and 52.2%, respectively, and the stage of discovery is relatively low. Early lesions have a significantly better prognosis. Especially for stage I non-small cell lung cancer, the 5-year survival rate after radical surgical resection can reach 58-73%. Therefore, one of the most effective ways to improve the prognosis of lung cancer is to further increase the early diagnosis rate of lung cancer.

At present, the detection of lung cancer mainly relies on chest X-ray or CT examination. A large-scale randomized controlled lung cancer screening study involving 53,454 subjects conducted in the United States showed that low-dose spiral CT screening can detect more early lung cancers, and compared with chest X-ray screening, the mortality rate of lung cancer is lower. 20% lower. However, this screening method also has many problems: first, the false positive rate of positive findings in imaging is as high as 95%, and how to make further identification of these positive findings is still a big problem; in addition, low-dose helical CT examinations have the problem of overdiagnosis, leading to some unnecessary examinations and even invasive treatments; and repeated imaging examinations have the risk of radiation-induced tumors. Another study concluded that although low-dose spiral CT screening for lung cancer can reduce its mortality, the cost is relatively expensive, which is not conducive to popularization and application. Therefore, there is still an urgent need for a non-invasive method to assist in the screening and early diagnosis of tumors.

Human blood is rich in various cellular components and molecular substances, which can well reflect the physiological and pathological states of different tissues and organs in the body, and its specimens are easy to obtain. Therefore, it is an ideal non-invasive diagnosis method for tumors. Therefore, markers with good sensitivity and specificity remain to be developed.

Contents of the invention

The purpose of the present invention is to provide a kit for auxiliary diagnosis of patients with non-small cell lung cancer based on the protein marker PAK4.

The present invention claims the application of the product for detecting the protein marker PAK4 in the preparation of a kit for auxiliary diagnosis of patients with non-small cell lung cancer. If the content of the protein marker PAK4 in the plasma of the examinee is above 1.976ng/ml, the examinee is a candidate non-small cell lung cancer patient; if the content of the protein marker PAK4 in the plasma of the examinee is less than 1.976ng/ml, then The subjects were candidate non-small cell lung cancer patients.

The non-small cell lung cancer patient may be a lung phosphocarcinoma patient. If the content of the protein marker PAK4 in the plasma of the examinee is above 1.885ng/ml, the examinee is a candidate lung phosphocarcinoma patient; if the content of the protein marker PAK4 in the plasma of the examinee is less than 1.885ng/ml, the examinee Those who are candidates for non-pulmonary phosphorus cancer patients.

The non-small cell lung cancer patient may be a lung adenocarcinoma patient. If the content of the protein marker PAK4 in the plasma of the candidate is above 1.976ng/ml, the candidate is a candidate lung adenocarcinoma patient; if the content of the protein marker PAK4 in the plasma of the candidate is less than 1.976ng/ml, the candidate for the test Candidate non-lung adenocarcinoma patients.

The non-small cell lung cancer patient may be a stage I non-small cell lung cancer patient. If the content of the protein marker PAK4 in the plasma of the examinee is above 1.662ng/ml, the examinee is a candidate for stage I non-small cell lung cancer patients; if the content of the protein marker PAK4 in the plasma of the examinee is less than 1.662ng/ml , the examinee is a candidate non-stage I non-small cell lung cancer patient.

The invention also protects a kit for auxiliary diagnosis of patients with non-small cell lung cancer, including a product for detecting the protein marker PAK4. The kit can also have a carrier recorded with the following judgment criteria: if the content of the protein marker PAK4 in the plasma of the examinee is above 1.976 ng/ml, the examinee is a candidate non-small cell lung cancer patient; if the examinee The content of the protein marker PAK4 in plasma is less than 1.976ng/ml, and the candidate is a candidate non-non-small cell lung cancer patient.

The non-small cell lung cancer patient may be a lung phosphocarcinoma patient. The kit can also have a carrier recorded with the following judgment criteria: if the content of the protein marker PAK4 in the plasma of the examinee is above 1.885 ng/ml, the examinee is a candidate lung cancer patient; The content of the protein marker PAK4 in the medium is less than 1.885ng/ml, and the candidate is a candidate non-pulmonary phosphorus cancer patient.

The non-small cell lung cancer patient is a lung adenocarcinoma patient. The kit can also have a carrier recorded with the following judgment criteria: if the content of the protein marker PAK4 in the subject's plasma is 1.976 ng/ml or more, the subject is a candidate lung adenocarcinoma patient; if the subject's plasma The content of the protein marker PAK4 in the medium is less than 1.976ng/ml, and the candidate is a candidate non-lung adenocarcinoma patient.

The non-small cell lung cancer patient may be a stage I non-small cell lung cancer patient. The kit can also have a carrier with the following judgment criteria: if the content of the protein marker PAK4 in the plasma of the subject is 1.662 ng/ml or more, the subject is a candidate stage I non-small cell lung cancer patient; The content of the protein marker PAK4 in the plasma of the examinee is less than 1.662ng/ml, and the examinee is a candidate non-stage I non-small cell lung cancer patient.

Any of the above-mentioned products for detecting protein marker PAK4 can be a kit for detecting protein marker PAK4, specifically "p21 protein-activated kinase 4 (PAK4) detection kit (enzyme-linked immunosorbent assay)" ", purchased from "USCN LifeScience, Wuhan, China", the article number is "SEH464Hu".

The invention also protects the application of the product for detecting the protein marker PAK4 in the preparation of a kit for judging the prognosis of patients with non-small cell lung cancer. The stronger the expression of the protein marker PAK4 in the tumor tissue, the worse the prognosis of the patient. The product used to detect the protein marker PAK4 may be an antibody to the protein marker PAK4, specifically a rabbit-derived polyclonal PAK4 antibody, purchased from abnova.

The full name of the protein marker PAK4 is p21-activated kinase 4 (p21-activated kinase4, serine/threonine-proteinkinase PAK4), which is one of the serine/threonine protein kinases. It was first discovered by Abo et al. in 1998 and can induce filopodia Formation and polymerization of actin, leading to remodeling of the cytoskeleton. The gene encoding the protein marker PAK4 is located on chromosome 19q13.2-13.3. In the present invention, the inventors discovered the existence of the protein marker PAK4 in the peripheral blood of NSCLC patients for the first time, and found that the concentration of the protein marker PAK4 in the peripheral blood of NSCLC patients was higher than that of healthy people, that is, the protein marker PAK4 has an auxiliary diagnosis Value for NSCLC patients. In the present invention, the inventors found through immunohistochemistry that the expression of the protein marker PAK4 in the tumor tissue of NSCLC patients was significantly higher than that in the normal tissue next to the tumor tissue, and the expression intensity was related to the prognosis and survival time, which can be used as a candidate prognostic factor. The invention has great value for auxiliary diagnosis and prognosis of non-small cell lung cancer.

Description of drawings

Fig. 1 is the ROC curve in embodiment 1.

Fig. 2 is the ROC curve in embodiment 2.

Fig. 3 is the ROC curve in embodiment 3.

Fig. 4 is the ROC curve in embodiment 4.

Fig. 5 is the partial immunohistochemical photograph obtained in embodiment 5.

Figure 6 shows the relationship between the expression intensity of the protein marker PAK4 and the prognosis in 154 patients with non-small cell lung cancer.

detailed description

The following examples facilitate a better understanding of the present invention, but do not limit the present invention. The experimental methods in the following examples are conventional methods unless otherwise specified. The test materials used in the following examples, unless otherwise specified, were purchased from conventional biochemical reagent stores. Quantitative experiments in the following examples were all set up to repeat the experiments three times, and the results were averaged.

Specimens were collected and preserved in accordance with the recommendations provided by the Human Proteome Organization (HUPO) and the Early Detection Research Network (EDRN). All participants provided written informed consent.

Statistical analysis was completed by GraphpadPrism5 software (SanDiego, CA, USA) and SPSS15.0 software (IBM, Chicago, USA). The differences among groups of protein markers in plasma were analyzed by Mann-Whitney U test. The difference of immunohistochemical staining between tumor tissue and normal tissue was tested by Wilcoxon rank sum test. The Kaplan-Meier method was used for survival analysis, the Log-rank test was used for survival differences between groups, and the Cox proportional hazards model was used for multivariate analysis. A P value ≤ 0.05 was considered statistically significant.

Example 1. Application of the protein marker PAK4 in the auxiliary diagnosis of patients with non-small cell lung cancer

The patients used in this embodiment were 153 clinically diagnosed non-small cell lung cancer patients (peripheral venous blood was obtained before surgery, and the patients had not received chemotherapy, radiotherapy, intervention, etc.) and 108 healthy people (healthy volunteers) , lung cancer or other diseases were ruled out before blood collection), all volunteers with informed consent.

Blood samples were collected in EDTA anticoagulant tubes, and centrifuged at 3000 rpm for 20 minutes within 1 hour of blood collection to obtain plasma, which was frozen at -80°C. After dissolving at room temperature, Jiang Ce detected the concentration of protein marker PAK4, protein marker Cyfra21-1, protein marker SCC and protein marker CEA in plasma respectively.

The full name of the kit for detecting the protein marker PAK4 is "p21 Protein-Activated Kinase 4 (PAK4) Detection Kit (Enzyme-Linked Immunosorbent Assay)", purchased from "USCN LifeScience, Wuhan, China", and the article number is "SEH464Hu". The full name of the kit used to detect the protein marker Cyfra21-1 is " CYFRA21-1)", purchased from "Roche Applied Chemicals Company", the article number is "11820966". The full name of the kit for detecting the protein marker SCC is "i2000SRSCC detection reagent", purchased from "Abbott Laboratories", the article number is "8D18 ". The full name of the kit used to detect the protein marker CEA is " Carcinoembryonicantigen (CEA)" was purchased from "Roche Applied Chemicals" with the article number "11731629".

The concentration results of each protein marker in the plasma of 153 patients with non-small cell lung cancer are shown in Table 1. The results of the concentration of each protein marker in the plasma of 108 healthy people are shown in Table 2.

Table 1 Concentration of each protein marker in the plasma of patients with non-small cell lung cancer (+ represents positive diagnosis, - represents negative diagnosis)

Table 2 Concentration of each protein marker in healthy human plasma (+ represents positive diagnosis, - represents negative diagnosis)

Draw the ROC (receiveroperatingcharacteristiccurve) curve (see Figure 1).

In the diagnosis of 153 NSCLC patients and 108 healthy controls: the area under the ROC curve (AUC, area under the curve) of PAK4 is 0.985 (95% CI is 0.962-0.996), if the concentration of PAK4 at the maximum Youden index is selected as the cut-off value (1.976ng/ml), its sensitivity was 88.24%, its specificity was 100%, Youden's index was 0.8824; the AUC value of Cyfra21-1 was 0.837 (95% CI was 0.787-0.880), and the critical value was 3.3ng/ml ml, its sensitivity, specificity and Youden index were 50.33%, 94.44% and 0.4477 respectively; the AUC value of SCC was 0.569 (95% CI was 0.507-0.630), and when the critical value was 1.5ng/ml, its sensitivity , specificity and Youden index were 28.10%, 95.37% and 0.2347 respectively; the AUC value of CEA was 0.711 (95% CI was 0.652-0.766), and when the critical value was 5ng/ml, the sensitivity, specificity and Youden index were respectively are 29.41%, 99.07%, and 0.2755. The AUC value of PAK4 was significantly higher than that of Cyfra21-1, SCC and CEA (p<0.01), indicating that PAK4 was more effective in the diagnosis of NSCLC than Cyfra21-1, SCC and CEA.

Example 2. Application of the protein marker PAK4 in the auxiliary diagnosis of patients with stage I non-small cell lung cancer

The patients used in this embodiment were 27 clinically diagnosed patients with stage I non-small cell lung cancer (peripheral venous blood was obtained before surgery, and the patients had not received chemotherapy, radiotherapy, intervention, etc.) and 35 healthy people (healthy Volunteers, lung cancer or other diseases have been ruled out before blood collection), all volunteers with informed consent.

Blood samples were collected in EDTA anticoagulant tubes, and centrifuged at 3000 rpm for 20 minutes within 1 hour of blood collection to obtain plasma, which was frozen at -80°C. After dissolving at room temperature, Jiang Ce detected the concentration of protein marker PAK4, protein marker Cyfra21-1 and protein marker CEA in plasma respectively.

The full name of the kit for detecting the protein marker PAK4 is "p21 Protein-Activated Kinase 4 (PAK4) Detection Kit (Enzyme-Linked Immunosorbent Assay)", purchased from "USCN LifeScience, Wuhan, China", and the article number is "SEH464Hu". The full name of the kit used to detect the protein marker Cyfra21-1 is " CYFRA21-1)", purchased from "Roche Applied Chemicals Company", the article number is "11820966". The full name of the kit for detecting protein marker CEA is " Carcinoembryonicantigen (CEA)" was purchased from "Roche Applied Chemicals" with the article number "11731629".

The concentration results of each protein marker in the plasma of 27 clinically diagnosed patients with stage I non-small cell lung cancer are shown in Table 3. The results of the concentration of each protein marker in the plasma of 35 healthy people are shown in Table 2.

Table 3 Concentration of each protein marker in plasma of patients with stage I non-small cell lung cancer (+ represents positive diagnosis, - represents negative diagnosis)

Table 4 Concentration of each protein marker in healthy human plasma (+ represents positive diagnosis, - represents negative diagnosis)

Draw the ROC (receiveroperatingcharacteristiccurve) curve (see Figure 2).

In the diagnosis of 27 patients with stage I NSCLC and 35 healthy controls: the area under the ROC curve (AUC, area under the curve) of PAK4 was 0.989 (95% CI was 0.922-0.996), if the concentration of PAK4 at the maximum Youden index was selected as At the critical value (1.662ng/ml), the sensitivity was 96.30%, the specificity was 94.29%, and the Youden index was 0.9059; the AUC value of Cyfra21-1 was 0.711 (95% CI was 0.582-0.819), and the critical value was 3.3 ng/ml, its sensitivity, specificity and Youden index were 29.63%, 88.57% and 0.182; The specificity, specificity and Youden's index were 11.11%, 100% and 0.1111, respectively. The AUC value of PAK4 was significantly higher than that of Cyfra21-1 and CEA (p<0.01), indicating that the diagnostic efficiency of PAK4 for stage I NSCLC was better than that of Cyfra21-1 and CEA.

Example 3. Application of the protein marker PAK4 in the auxiliary diagnosis of lung adenocarcinoma

The patients used in this embodiment were 64 clinically diagnosed lung adenocarcinoma patients (peripheral venous blood was obtained before surgery, and the patients had not received chemotherapy, radiotherapy, intervention, etc.) and 108 healthy people (healthy volunteers, Lung cancer or other diseases have been ruled out before blood collection), all volunteers with informed consent.

Blood samples were collected in EDTA anticoagulant tubes, and centrifuged at 3000 rpm for 20 minutes within 1 hour of blood collection to obtain plasma, which was frozen at -80°C. After dissolving at room temperature, Jiang Ce detected the concentration of protein marker PAK4, protein marker Cyfra21-1 and protein marker CEA in plasma respectively.

The full name of the kit for detecting the protein marker PAK4 is "p21 Protein-Activated Kinase 4 (PAK4) Detection Kit (Enzyme-Linked Immunosorbent Assay)", purchased from "USCN LifeScience, Wuhan, China", and the article number is "SEH464Hu". The full name of the kit used to detect the protein marker Cyfra21-1 is " CYFRA21-1)", purchased from "Roche Applied Chemicals Company", the article number is "11820966". The full name of the kit for detecting protein marker CEA is " Carcinoembryonicantigen (CEA)" was purchased from "Roche Applied Chemicals" with the article number "11731629".

The concentration results of each protein marker in the plasma of 64 clinically diagnosed patients with lung adenocarcinoma are shown in Table 5. The results of the concentration of each protein marker in the plasma of 108 healthy people are shown in Table 6.

Table 5 Concentration of each protein marker in plasma of patients with lung adenocarcinoma (+ represents positive diagnosis, - represents negative diagnosis)

Table 6 The concentration of each protein marker in the plasma of healthy people (+ represents positive diagnosis, - represents negative diagnosis)

Draw the ROC (receiveroperatingcharacteristiccurve) curve (see Figure 3).

In the diagnosis of 64 lung adenocarcinoma patients and 108 healthy controls: the area under the ROC curve (AUC, area under the curve) of PAK4 was 0.987 (95% CI was 0.957-0.998), if the concentration of PAK4 at the maximum Youden index was selected as At the critical value (1.976ng/ml), the sensitivity was 90.62%, the specificity was 100%, and the Youden index was 0.9062; the AUC value of Cyfra21-1 was 0.745 (95% CI was 0.673-0.808), and the critical value was 3.3 ng/ml, its sensitivity, specificity and Youden index were 32.81%, 95.37% and 0.2818; The specificity, specificity and Youden index were 45.31%, 99.07% and 0.4438 respectively. The AUC value of PAK4 was significantly higher than that of Cyfra21-1 and CEA (p<0.01), indicating that the diagnostic efficiency of PAK4 for lung adenocarcinoma was better than that of Cyfra21-1 and CEA.

Example 4. Application of the protein marker PAK4 in the auxiliary diagnosis of pulmonary phosphocarcinoma

The patients used in this embodiment were 89 patients with clinically diagnosed pulmonary phosphocarcinoma (peripheral venous blood was obtained before surgery, and the patients had not received chemotherapy, radiotherapy, intervention, etc.) and 108 healthy people (healthy volunteers, Lung cancer or other diseases have been ruled out before blood collection), all volunteers with informed consent.

Blood samples were collected in EDTA anticoagulant tubes, and centrifuged at 3000 rpm for 20 minutes within 1 hour of blood collection to obtain plasma, which was frozen at -80°C. After dissolving at room temperature, Jiang Ce detected the concentration of protein marker PAK4, protein marker Cyfra21-1 and protein marker CEA in plasma respectively.

The full name of the kit for detecting the protein marker PAK4 is "p21 Protein-Activated Kinase 4 (PAK4) Detection Kit (Enzyme-Linked Immunosorbent Assay)", purchased from "USCN LifeScience, Wuhan, China", and the article number is "SEH464Hu". The full name of the kit used to detect the protein marker Cyfra21-1 is " CYFRA21-1)", purchased from "Roche Applied Chemicals Company", the article number is "11820966". The full name of the kit for detecting the protein marker SCC is "i2000SRSCC detection reagent", purchased from "Abbott Laboratories", the article number is "8D18 ". The full name of the kit used to detect the protein marker CEA is " Carcinoembryonicantigen (CEA)" was purchased from "Roche Applied Chemicals" with the article number "11731629".

The concentration results of each protein marker in the plasma of 89 patients with clinically diagnosed pulmonary phosphocarcinoma are shown in Table 7. The results of the concentration of each protein marker in the plasma of 108 healthy people are shown in Table 8.

Table 7 The concentration of each protein marker in the plasma of patients with lung phosphocarcinoma (+ represents positive diagnosis, - represents negative diagnosis)

Table 8 The concentration of each protein marker in the plasma of healthy people (+ represents positive diagnosis, - represents negative diagnosis)

Draw the ROC (receiveroperatingcharacteristiccurve) curve (see Figure 4).

In the diagnosis of 89 lung squamous cell carcinoma patients and 108 healthy controls: the area under the ROC curve (AUC, area under the curve) of PAK4 was 0.984 (95% CI was 0.954-0.996), if the concentration of PAK4 at the maximum Youden index was selected as At the critical value (1.885ng/ml), the sensitivity was 88.76%, the specificity was 98.15%, and the Youden index was 0.8691; the AUC value of Cyfra21-1 was 0.903 (95% CI was 0.853-0.941), and the critical value was 3.3 ng/ml, its sensitivity, specificity and Youden index were 62.92%, 94.44% and 0.5736; The sensitivity, specificity and Youden index were 46.07%, 95.37% and 0.4144 respectively; the AUC value of CEA was 0.681 (95% CI was 0.612-0.746), and when the cut-off value was 5ng/ml, the sensitivity, specificity and Youden index The indices are 17.98%, 99.07% and 0.1705 respectively. The AUC value of PAK4 was significantly higher than that of Cyfra21-1, SCC and CEA (p<0.01), indicating that the diagnostic efficiency of PAK4 for lung squamous cell carcinoma was better than that of Cyfra21-1, SCC and CEA.

Example 5, Expression Intensity of Protein Marker PAK4 in Tumor Tissue and Normal Lung Tissue of NSCLC Patients

154 patients with clinically diagnosed non-small cell lung cancer (volunteers who gave informed consent), respectively took tumor tissues and normal lung tissues to make paraffin sections and performed immunohistochemistry. The primary antibody for immunohistochemical staining was polyclonal PAK4 antibody derived from rabbit, which was purchased from abnova (PAB-2301, diluted 1:70).

Some photos are shown in Figure 5. The protein marker PAK4 is mainly expressed in the cytoplasm of tumor cells, but rarely expressed in normal tissues or rarely expressed in respiratory bronchiolar epithelial cells. In NSCLC, the positive expression of protein marker PAK4 in lung tumor tissue was significantly increased (81.2%), which was higher than that in corresponding normal lung tissue (11.0%), and the difference was statistically significant (p<0.001).

Staining scores under the microscope were independently completed by two experienced pathologists in a single-blind situation. They were divided into 0, 1, 2, and 3 grades according to the staining intensity, and 0 (1-5% according to the number of stained cells). ), 1 (6-25%), 2 (26-50%), 3 (51-75%) and 4 (76-100%). The final score is based on the comprehensive evaluation of the staining intensity and the number of positive cells, and is divided into - (negative), + (weak positive, 1-4 points), ++ (positive, 5-8 points) and +++ (strong positive, 8 points). points or more). The relationship between the expression intensity of the protein marker PAK4 and the prognosis in 154 patients with non-small cell lung cancer is shown in Figure 6. In patients with survival time ≤ 5 years, the proportion of positive protein marker PAK4 expression (++/+++) was significantly higher than in patients > 5 years (39.5% vs 21.6%, p = 0.012). Kaplan-Meier survival analysis showed that the survival of patients with negative expression of protein marker PAK4 (n=29) and patients with weak positive expression (n=77) was better than that of patients with positive expression (n=48), (p=0.009,0.056 ). Among them, the 5-year survival rate of patients with negative protein marker PAK4 expression was 65.3%, the 5-year survival rate of weakly positive group was 50.1%, and the 5-year survival rate of positive group was 32.2%. The results suggest that the expression of the protein marker PAK4 is significantly increased in NSCLC, and the stronger the expression, the worse the prognosis, so it can be used as an independent factor to judge the prognosis of NSCLC (p=0.019, HR1.541, 95%CI1.073-2.211) .

Claims (5)

1. for detecting the application of product in the kit for the preparation of auxiliary diagnosis Patients with Non-small-cell Lung of protein marker PAK4.

2. apply as claimed in claim 1, it is characterized in that: described Patients with Non-small-cell Lung is lung phosphorus cancer patient.

3. apply as claimed in claim 1, it is characterized in that: described Patients with Non-small-cell Lung is patients with lung adenocarcinoma.

4. apply as claimed in claim 1, it is characterized in that: described Patients with Non-small-cell Lung is I phase Patients with Non-small-cell Lung.

5. for the product that detects protein marker PAK4 for the preparation of the application carried out Patients with Non-small-cell Lung in the kit of Index for diagnosis.