| Objective: The morbidity and mortality of lung cancer are still high.Furthermore,some unhealthy eating habits,outside atmospheric environment pollution and occupational dusts and chemicals contact and the popularity of the tobacco industry make us more likely to get lung cancer,which constantly are threatening human health.Due to lung cancer has the high grade malignancy and the lack of specificity of clinical symptoms in the early stage,we can't discovery and treat it early,so as to make the delay of it.So that the majority of lung cancer patients have been diagnosed at the late stage,have to give priority to palliative symptomatic treatment,which greatly shorten their lifetime.Therefore,the diagnosis of lung cancer,especially early screening diagnosis is particularly important,which is important to prolong survival time and improve patient quality of life.Pathological diagnosis is still the gold standard in the diagnosis of lung cancer.Low dose CT is the major means of lung cancer screening.But due to a series of problems,such as high false positive rate,radiation effect,excessive diagnosis and high cost of inspection,it is still controversial whether patients really benefit from it.In recent years,serum lung cancer autoantibodies detection methods in the screening and diagnosis of lung cancer showed the good sensitivity and specificity,and the characteristic of its simple,non-invasive,promoting easily increase participants' adherence,for providing a new way of lung cancer diagnosis and screening.Inactivation of p53 protein caused by p53 mutation is an important step in cancer emerging.P53 autoantibodies are usually produced by mutations in the p53 gene product.PGP9.5 is a kind of ubiquitin hydrolase,which is expressed in nerve tissue,and is highly expressed in primary lung cancer and lung cancer cell lines.SOX2 is a transcription factor that induces tumor signalEGFR and BCL2L1,and promotes the proliferation and survival of lung cancer cells.GAGE 7 belongs to the tumor / testis antigen,which is merely expressed in malignant tumor and testis tissue,and can inhibit cell apoptosis.GBU4-5 belongs to the ATP binding RNA helicase,plays an important role in the process of carcinogenesis,and has tumor specificity and immunogenicity.MAGE A1 belongs to human melanoma antigen family,which only expressed in malignant tumors and testicular tissue,and may has relationship with gene transcriptional regulation and transformation or progress of cancer.CAGE belongs to the family of DEAD box helicase,and its expression is related to cell cycle,which activates ERK and p38 protein in cancer cells and makes tumor cells proliferate.This paper collected serum samples from patients with different types,and applied enzyme linked immunosorbent assay(ELISA)to detect seven serum autoantibodies(p53,PGP9.5,SOX2,GAGE7,GBU4-5,MAGEA1,CAGE)level of the patients.We observed sensitivity and specificity of this detection method in lung cancer screening,compared the difference of the levels of autoantibody detection between groups,and discussed the function of autoantibody detection in diagnosis and screening of lung cancer.Methods:From November 2016 to February 2017,we recorded 140 patients in the fourth hospital of hebei medical university.88 cases were as the lung cancer group,which were diagnosed as lung cancer by pathological examination,owning clear histological type and TNM staging of lung cancer,and excluded from other organ malignant tumor;47 cases were as the control group,which excluded with imaging or pathology examination from malignant tumor lung and other organs(including 34 cases of benign lung disease patients and 13 healthy subjects).1 case with vascular immune maternal T cell lymphoma,2cases with hamartoma,1 case with teratoma,1 case with sarcoma were confirmed by pathology.After collecting the whole blood of all patients,we collected the serum specimens by centrifugation.Then the serum specimens were numbered anddivided into two groups.We detected seven kinds of autoantibodies by ELISA method: the solid plates of kits were coated by seven purified antigens.Add respectively diluted serum samples into microporous enveloped by antigens;make autoantibodies in serum samples bind specifically with the antigen of microporous by the first incubation.Wash out uncombined samples,and add the enzyme tag anti-human IgG antibody(enzyme combination).In the second incubation,enzyme tag anti-human IgG antibody adsorbed on autoantibodies of the solid phase carrier,forming the Antigen antibody enzyme labeled antibody complex.Wash out enzyme uncombined IgG antibodies,and add Chromogenic substrate into microporous.After the reaction,we measured its absorbance by Microplate Reader under 450 nm wavelength,and calculate the relative concentrations of autoantibodies through standard curve.And according to the "positive judgment value",we judge detection of antibody concentration "negative" or "positive".The upper limit values of the seven autoantibodies were as follows: p53: 13.1U/ml;PGP9.5: 11.1U/ml;SOX2:10.3U/ml;GAGE7: 14.4U/ml;GBU4-5:7.0U/ml;MAGEA1: 11.9U/ml;CAGE:7.2U/ml.(Note: this data was validated by 2008 large sample multicenter clinical trials).Then calculate the sensitivity and specificity of this method and compare among antibody levels pf seven groups.We use the Mann-Whitney U test to measure difference between non-normal measurement data.Chi square test was used to compare the positive rates between groups.All tests were two-sided test,P<0.05 was statistically significant difference.Results:1 Clinical characteristics of the study:Lung cancer group included 88 cases of lung cancer patients as the research object.Including 56 cases of male patients,accounting for 63.6%,distributed in the age range from 35 to 80 years old,the average age was 61.2± 9.4 years,60 years of age or older in 55 cases,accounting for 62.5%;54cases with adenocarcinoma,accounted for 61.4%,18 cases with squamous cell carcinoma,accounting for 20.4%,16 cases with other types of lung cancer,accounting for 18.2%;42 cases with smoking the history,accounting for47.7%;53 cases of I and II patients with lung cancer,accounting for 60.2%,35 cases of stage III and IV patients with lung cancer,accounting for 39.8%.34 patients with benign lung disease were included in the study.Including 18 cases with male patients,accounting for 52.9%;age range distribution from 25 to 86 years old,the average age was 54.8 ± 16.2 years,11 cases with 60 years of age or older,accounting for 32.4%;10 patients with smoking history,accounting for 29.4%;16 pneumonia patients,accounted for47%;9 cases with pulmonary tuberculosis,accounting for 26.5%,5 cases with chronic obstructive pulmonary disease,accounting for 14.7%,4 cases of other types of patients,accounting for 11.8%.13 healthy subjects were included in the study.Including 6 cases of male patients,accounting for 46.2%;age range distribution from 39 to 76 years old,the average age was 56.5 ± 11.5 years,4 cases were older than 60 years old,accounting for 30.8%.There were 1 case of patients with vascular immune maternal T cell lymphoma,2 case of hamartoma,1 case of teratoma,and 1 case with sarcoma.2 Compare the positive rate of seven autoantibodies in lung cancer group and control group:The positive rates of p53 and GBU4-5 antibody in lung cancer group were higher than those in control group(p53:17.0% vs 4.3%,P<0.05;GBU4-5:15.9% vs 0%,P<0.05).There was no significant difference in the positive rate between the five other autoantibodies in lung cancer group and control group(P>0.05).3 The sensitivity,specificity,diagnostic coincidence rate and correct index of the seven kinds of autoantibodies in the diagnosis of lung cancer:Sensitivity: 9.1%-17%;specificity: 89.4%-100%;diagnostic coincidence rate: 38.5%-45.2%;correct index: 0.03-0.16.4 The sensitivity,specificity,diagnostic coincidence rate and correct index of combined detection of autoantibodies in the diagnosis of lung cancer:The sensitivity of combined detection of p53 and GBU4-5 antibody was27.2%,specificity was 95.7%,the diagnostic coincidence rate was 51.1%,the correct index is 0.23;the sensitivity of combined detection of p53,GBU4-5and MAGEA1 was 34.1%,specificity was 91.5%,the diagnostic coincidence rate was 54.1%,the correct index is 0.26;the sensitivity of combined detection of seven kinds of autoantibodies was 46.6%,the specificity was70.2%,the diagnostic coincidence rate was 54.8%,the correct index was 0.17.5 The sensitivity and specificity of combined detection of seven kinds of antibodies in the diagnosis of different types of lung cancer:The sensitivity and specificity of the seven antibodies in the diagnosis of lung adenocarcinoma was 35.2%,and the specificity was 70.2%.The sensitivity and specificity for the diagnosis of lung squamous cell carcinoma was 72.2% and the specificity was 70.2%.6 Comparison of the expression levels of the seven autoantibodies between lung cancer group and control group:The expression level of p53 antibody in lung cancer group was higher than that in control group(2.30(0.83,7.18)vs1.60(0.20,2.80),P<0.05).There was no significant difference in the expression level of the other six autoantibodies in lung cancer group and control group(P>0.05).7 Comparison of the expression levels of the seven autoantibodies in different pathological types lung cancer of lung cancer group and control group:7.1 Comparison of serum levels of seven autoantibodies between patients with lung adenocarcinoma and lungsquamous cell carcinoma:The expression level of PGP9.5 antibody in patients with lung squamous cell carcinoma was higher than that in lung adenocarcinoma(5.05(2.13,19.85)vs2.45(1.60,3.73),P<0.05).There was no significant difference in the expression level of the other six autoantibodies in lung adenocarcinoma and squamous cell carcinoma(P>0.05).7.2 Comparison of serum levels of seven autoantibodies in patients between lung adenocarcinoma and control group:There was no significant difference in the expression level of seven autoantibodies between lung adenocarcinoma patients and controls(P>0.05).7.3 Comparison of serum levels of seven kinds of autoantibodies in patients between squamous cell carcinoma of lung and control group:The expression level of p53,PGP9.5 and MAGEA1 antibody in serum of patients between lung squamous cell carcinoma were higher than the control group,the difference was statistically significant(p53:2.95(1.63,19.70)vs1.60(0.20,2.80),P<0.05;PGP9.5:5.05(2.13,19.85)vs2.60(1.90,4.00),P<0.05;MAGEA1:2.15(0.20,13.58)vs0.50(0.10,1.70),P<0.05).There was no significant difference in the expression level of the other four autoantibodies in patients with squamous cell carcinoma of the lung(P>0.05).8 Comparison of the levels of serum autoantibodies in patients with stage I,II lung cancer and stage III,IV lung cancer:The expression level of patients with stage III and IV lung cancer patients serum PGP9.5,GBU4-5 and CAGE antibodies were higher than that in stage I,II lung cancer,the difference was statistically significant(PGP9.5:3.10(2.30,18.10)vs2.30(1.55,3.75),P<0.05;GBU4-5:1.50(0.20,22.00)vs0.40(0.00,1.15),P<0.05;CAGE:1.70(1.20,8.10)vs1.40(0.20,2.05),P<0.05).There was no significant difference in the expression level of the other four autoantibodies in patients with stage I,II and stage III,IV(P>0.05).Conclusions:1 Detection of single antibody(p53,PGP9.5,SOX2,GAGE7,GBU4-5,MAGEA1,CAGE)of lung cancer screening high specificity(89.4%-100%),low sensitivity(9.1%-17.0%),still can not meet the indicators for lung cancer screening.2 Two(p53 and GBU4-5)or three(p53,GBU4-5,and MAGEA1)lung cancer autoantibodies in combined detection of single antibody can improve the sensitivity of detection.3 The sensitivity of combined detection of seven kinds of serum autoantibodies in lung cancer screening was 46.6%,and the specificity was70.2%.It is recommended that the combined detection of the sevenautoantibodies can be used in the clinical screening of lung cancer.4 P53,PGP9.5 and MAGEA1 antibodies can be used as the differential markers between lung squamous cell carcinoma and benign lung disease.5 The expression levels of PGP9.5,GBU4-5 and CAGE were correlated with tumor burden. |
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