| Background and ObjectivesCurrently, the incidence and mortality of lung cancer are raising, lung cancer becomes the most common malignant tumor in the world. In the present society, with the rapid development of industrial, environment pollution problems increasingly. No doubt it will bring with it the means for prolonging life through improved medical care and living standards. The progress achieved in social development had also generated new problems, such as an ageing population. For these reasons, the incidence of lung cancer is rising yearly. Based on data from 2013, every year about1.5 million people die from lung cancer in the world, 5-year survival rate of the tumor is still low, only a quarter of people who have lung cancer survive for 5years after diagnosis. In China, Lung cancer is also the highest incidence of malignant tumor, the data shown at the China International Lung Cancer Seminar indicate that the population with lung cancer in China is the largest in the world, it is noteworthy that the incidence of lung cancer in China grows rapidly at 26.9% per year in recent years.Among which, non-small cell lung cancer(NSCLC) accounts for about 80% of all lung cancers, this disease leading to shorten the survival time and diminished quality of life for the patients. In recent years, human beings have indefatigable exploration about the pathogenesis, prevention, diagnosis and treatment of tumors. Therapeutic methods to malignant tumors has constantly improved and developed, novel agents and treatment approaches have yet to emerge, however it could only alleviate the recurrence and metastasis temporarily, and there has been an increasing trend in the mortality of malignant tumors. Therefore, it is particularly important to accurately determine the prognosis of each patient, and then take all necessary treatments in advance. The factor that effect prognostics is highly complex, now the pathological staging is still means to guide the conditions of prognosis, included tumor size,pleural metastasis, regional lymph node metastasis, distant metastasis and so on.However, according to the current TNM staging, it is not possible to explain why similarly staged patients have different prognoses. Recently, with the rapid development of molecular biology and genetics, people at an angle of molecular biology in to find out the factor that effect on prognosis, and discovered many essential elements of prognosis. And in fact, there is no unified view about prognostic indicator of lung cancer.Materials and Methods1 Specimen collection33 specimens of tumor tissues and para-cancerous tissues with complete clinical data were collected from Henan cancer hospital between December 2008 and January 2010.2 Specimen processing All the lung cancer tissues and para-cancerous tissues were put in enzyme free freezing deposit tube, and preserved it at-80℃ refrigerator, and for the m RNA and protein extraction.3 Methods3.1 At first we through literature reviewing, summarizing and deep analysising and make a deeply excavation of GWAS to screening candidate genes. Such as CD133、CD24、CD34、OCT4、SOX3、PLAGL2、CD176、SALL4、CD105、TAZ、KIT、ABCG2、ALDH1、NANOG、SOX2、TERT, among them CD133 has been verified3.2 Extraction and detection of m RNA Firstly use Trizol to extract the total RNA from 10 specimens of lung cancer tissues and para-cancerous tissues, next the RNA was used to convert to synthesize first-strand c DNA, the last detected the expression of candidate genes by real time PCR. Analysis and screen the gene which have a difference in expression between in tumor tissue and in non-tumor tissue.3.3 Extraction and detection of m RNA A second time, we verified the selected gene in a large sample. Firstly use Trizol to extract the total RNA from lung cancer tissues and para-cancerous tissues, next the RNA was used to convert to synthesize first-strand c DNA,the last detected the expression of candidate genes by real time PCR. Once again analysis and screen the gene which have a difference in expression between in tumor tissue and in non-tumor tissue, and then analysis its correlations to clinicopathologic features.3.4 Extraction and detection of protein First, proteins are extracted from lung cancer tissues and para-cancerous tissues. Second, measured the selected gene at protein level by means of Western blotting. The relationship of each expressions and the correlation between the expressions and clinical pathological features and prognosis were analyzed.4 Statistics analysis Statistical analysis was executed by SPSS9.0 software, t-test was used to analyze the difference of m RNA relative expression quantity between lung cancer tissues and para-cancerous tissues. Then detected these proteins by western blot.Kaplan-Meier was performed on survival analysis, and a value less than 0.05 was considered as significance.Results1 The expression of m RNA in tissues In 10 cases lung cancer tissues, the expression of 16 genes: the relative expression quantity of ABCG2,ALDH1,CD34,CD24,CD105,CD133,CD176,OCT4,PLAG2,SALL4,SOX3,TAZ in lung cancer tissues is higher than para-cancerous tissues. The relative expression quantity of NANOG,KIT,SOX2 in lung cancer tissues is lower than para-cancerous tissues. The relative expression of CD24 m RNA in lung cancer tissues is 3.15 folds compared to para-cancerous tissues, p=0.002.2 The expression of CD24 m RNA in tissues and the relationship with clinicopathologic features2.1 In 33 samples, the expression of CD24 m RNA in lung cancer tissues is significantly higher than the adjacent normal tissues, the relative expression is 2.62 folds compared to para-cancerous tissues, p=0.0019.2.2 It indicated that the the relative expression of CD24 m RNA was no correlated with ages, p=0.52; The expression of CD24 m RNA in the different genders, pathological types degree of differentiation, TNM stages and the metastasis of lymph nodes was no significant difference, P values were 0.56, 0.16, 0.99, 0.76, 0.55.3 The relationship between the expression of CD24 m RNA and prognosis3.1 The results indicate that the relative expression of CD24 m RNA is negatively correlated with Progression-free survival(PFS), Pearson’s product-moment correlation coefficient r=-0.47,R2=0.221,p=0.0058.3.2 The relative expression of CD24 m RNA is negatively correlated with overall survival(OS), Pearson’s product-moment correlation coefficient r=-0.4653,R2=0.216,P=0.006.4 The expression of CD24 protein in tissues and the relationship with clinicopathologic features4.1 The expression of CD24 protein in lung cancer tissues is higher than the para-cancerous tissues, CD24 expression of lung cancer in protein was 2.46 times to para-cancerous tissues, p=0.0034.4.2 It indicated that the the expression of CD24 protein was no correlated with ages, p=0.37; The expression of CD24 protein in the different genders,pathological types degree of differentiation, TNM stages and the metastasis of lymph nodes was no significant difference, P values were0.65, 0.16, 0.58,0.74, 0.26.5 The relationship between the expression of CD24 protein and prognosis5.1 The results indicate that the relative expression of CD24 protein is negatively correlated with PFS, Pearson ’ s product-moment correlation coefficient r=-0.36, 95% confidence interval(CI) =-0.63 to 0.02, p=0.04.5.2 The expression of CD24 protein is negatively correlated with OS, Pearson’s product-moment correlation coefficient r=-0.3937, CI=-0.65 to-0.06,p=0.02.5.3 The median survival time in the CD24 high expression group was 46.43 months, the median survival time in the CD24 middle expression group was50.83 months, the median survival time in the CD24 low expression group was 69.67 months, p=0.0005.Conclusions1 CD24 m RNA is over-expressed in lung cancer tumor tissues, which is negatively correlated with the PFS and OS.2 CD24 protein is over-expressed in lung cancer tumor tissues, which is negatively correlated with the PFS and OS. CD24 low expression can lengthen a patient’s life cycle.3 CD24 can be used as a potential marker to estimate the prognosis of patients. |
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