| 1BackgroundThe incidence of lung cancer mortality is increased year by year. What is a enormous problem each pneumologue face to. The incidence of lung cancer in recent decades rose4times in our country.The incidence of lung cancer in Beijing increased60%in the last ten years.most cancer is not detected early and the rising incidence, it remains be the leading cause of cancer death. Non small cell lung cancer, in (a) the majority, is not sensitive to chemotherapy and radiation treatment.So, we choose surgical therapy firstly when we could. But the majority of patients with lung cancer already missed the operation time when the cancer is found.So the more research on lung cancer, improving the early diagnosis rate, looking for a better targeted treatment method are very necessary. The reasons of tumor formation are cell proliferation and differentiation, apoptosis, abnormal, the suppression of cell death. Abnormal apoptosis plays an important role in tumor occurrence, development. Expression of apoptosis inhibiting gene, apoptosis promoting gene and apoptosis effect of genes up-regulated and down-regulated, may break the balance of cell death and survival. When the balance breaks, abnormal cell proliferation and death will cause the tumor. In recent years, the researchs on apoptosis related gene is growing, but most of then are still in the stage of laboratory research. The patients could not enjoy the great benefit of it. Improving cancer diagnosis rate, reduced tumor mortality require us to do more about tumor apoptosis related genes. The non-operation treatment (such as radiotherapy, chemotherapy) principle is mainly induces apoptosis, but it has no obvious purpose, at the same time of killing of tumor cells, it inevitably kills normal human cells, so that the side effects are large, and can not be tolerated. Tumor occurrence, development undoubtedly influenced by a variety of gene regulation, mutual influence, mutual association. This paper mainly studies the relationship of Bax and caspase-3with lung cancer, and the relationship between then. Bax is apoptosis-promoting genes, at present, the researches about it and lung cancer are not uncommon, but most of then are researches about the single gene, or researches about bax and Bcl-2. But the study on caspase-3and bax at home and abroad is very rare.2Meterials and Methods2.1Object of studyIn this experiment, we choose non-small cell lung cancer specimens randomly of2009-2011after surgical resection in pathological Department of the1st Affiliated Hospital of Zhengzhou University. These are60patients, aged between the ages of36-87. Half of then are squamous cell carcinoma, and the ather are adenocarcinoma, all patients in the preoperative do not receive chemotherapy, and have definitive pathologic diagnosis. According to the United States of America TNM staging joint Cancer Classification Committee and the International Union against cancer developed staging criteria, another20cases in control group, as the adjacent normal tissue, all specimens were wax embedding, cuted into slices by professional personnel, each case with3pieces.2.2MethodsTo detect the expression of Bax and Caspase-3in lung cancer and normal tissues use immunohistochemical method (S-P method). To detect apoptotic cells use TUNEL method.2.3Standards of decision outcomesBax positive criteria:Cytoplasm or (and) the cell membrane appear brown granules, the first, the color and background staining showed no significant difference for0points, yellowish-brown, and background color difference,1, dyed brown, between Iand3points are2points, stain dark brown, with a background color difference is remarkable for3points, secondly, the stained cell ratio, high magnification (400times) of each field count of l00cells, each slice number5view, the number of positive cells in total cancer cell percentage.0%1,0%-10%2,10%-50%3,>50%for4points, the final score of first step and second step score results for the total value, total value of less than4for negative, greater than or equal to4as positive.Caspase-3positive standard:staining cytoplasm or (and) nuclei appear brown granules, accurate positioning and without background staining positive staining cytoplasm has brown granules of calm, more than the decision step with bax.2.4ApoptosisApoptosis interpretation standard of apoptotic cells after treatment by TUNEL method, the nucleus was stained brown, using optical microscopy, count of each of the1000tumor cells apoptosis cell number, statistical analysis.2.5statistical treatmentsAll statistics in the experiment are applied SPSS17.0software packet processing, the inspection level is a=0.05.The expression of Bax and Caspase-3and their relationships with clinical pathological characteristics use paired four tables chi-square test, in which the caspase-3expression and relationship of pleural effusions with continuity correction of the chi-square test (as expressed in pleural effusion were positive grid expected frequency of4.4, less than5). Looking for correlation of Bax and Caspase-3in non small cell lung cancer use paired data correlation analysis, its relation to apoptosis use linear correlation analysis.3Results3.1The expression of BaxBy immunohistochemistry, cell membrane and (or) the cytoplasm was stained brownish yellow,60cases of lung cancer, the positive rate was48.3%, adjacent normal tissue have20cases, the positive rate was60%, the statistical analysis, the difference was not statistically significant (x2=0.419, P=0.517), but the cancer tissue staining were deepper in cancer than normal tissue. The results show:the positive expression rate of high differentiated tissues (93.3%) was higher than the low differentiation (37.8%), the difference was statistically significant (X2=13.904, P=0.000). Lymph node metastasis positive rate (33.3%) was lower than those without lymph node metastasis (66.7%), the difference was statistically significant (X2=6.607, P=0.010). TNM stage I and II positive rate (61.5%) t was higher han that of III, IV positive rate (33.3%), the difference was statistically significant (X2=4.348, P=0.037). The expression of bax has no significant relationship with age, gender, pathological type, whether with pleural effusion, tumor diameter, whether smoking, tumor growth in the left lung and right lung showed (P>0.05).3.2The expression of caspase-3By immunohistochemistry, nuclei and (or) the cytoplasm was dyed brown,60cases of lung cancer, the positive rate was40%, adjacent normal tissue in20cases, the positive rate was40%, the statistical analysis, the difference was not statistically significant (X2=0.000, P=1.000). The results show, with lymph node metastasis positive rate (7.4%) was lower than those without lymph node metastasis (66.7%), the difference was statistically significant (X2=21.728, P=0.000). TNM stage I and II positive rate (56.4%) was higher than that of Ⅲ, Ⅳ positive rate (9.5%), the difference was statistically significant{X2=12.50, P=0.000). The expression of Caspase-3has no significant relationship with age, gender, pathological type, degree of differentiation, whether with pleural effusion, tumor diameter, whether smoking, tumor growth in the left lung and right lung (P>0.05).3.3The relation of Bax and Caspase-3in non-small cell lung cancerBax, as an apoptosis-promoting gene, caspase-3as an apoptosis effector gene, with other genes are related to apoptosis mediated by lung cancer cell apoptosis. The experimental results shown that, in lung cancer tissues, Bax was positive in31cases, caspase-3was positive in24cases, both of them were positive in22cases, and bax negative in29cases, caspase-3in36cases were negative, both of them expression and negative in26cases, application of paired data correlation analysis indicated, both have significant correlation, positive correlation.(r=0.593P=0.000))3.4The relationship of Bax, caspase-3and apoptosisBax and apoptosis were positively correlated,(r=0.855,P=0.000), caspase-3and apoptosis were positively correlated,(r=0.834,P=0.000). 4conclusions1When the expression of Bax, caspase-3was low, non small cell lung cancer with high grade malignancy and bad prognosis.2Bax and Caspase-3play a synergistic role in non small cell lung cancer occurrence, development.3Bax, caspase-3is closely related to apoptosis, repairing these genes is expected to promote apoptosis of tumor cells, inhibit tumor development. |
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