| Ovarian cancer is hackneyed gynecological sarcomata which can not be diagnosed out easily at the early period, however, can cause patients'death ultimately. The main reason why survival rate of the patients can't reach 30% within five years goes that 75% of these patients have been found involved in late ovarian cancer when they received the diagnosis. If the patients could be diagnosed in the early period of the disease, the survival rate for five years would reach 90%. Therefore, it is much of significance to carry out the research on diagnosis of the early period of ovarian cancer, in order to increase the survival rate.The current widely-used CA125, the biological marker of ovarian cancer found in 20 years ago, has been widely accepted for its diagnosis value. However, nearly 20% of the patients of ovarian cancer take on the low-level expression in CA125, and especially the CA125 shows negative at early period of ovarian cancer. The diagnosis rate of early period of ovarian cancer cannot still be up heaved when made reference to the enhancement of CA125, the unique norm of the diagnosis of ovarian cancer. It seems possible to improve the diagnosis rate of early period of ovarian cancer by combining CA125 and other biological markers of ovarian cancer. In this sense, great significance lies in researching the new biological markers. The current research, home and abroad, reveals that occurrence and development of ovarian cancer, to some extent, are related to some known or unknown genes. If we ascertain the variety of gene chart, and prove the nosogenesis of ovarian cancer, we will find it propitious to unfold the pathogeny of the disease, and supply the basic research for the tumor markers of ovarian cancer.The study of gene chip technology started from the 1990s. It's the research method which is combined with molecular biology, cytogenetics, biochemistry, bioinformatics and other high-techs. It's a large– scale, high-flux method for gene order and its functions. It's widely used in the study of tumor allied gene, the analysis of oncogene expression, the research of molecular tumor typing and tumor transgenation. In recent years, the gene chip technology is also applied to the study of ovarian cancer. It can analyze the features of overian cancer expression and find the early diagnosis molecular symbol, analyze the feature of drug resistant gene of ovarian cancer, judge the sensitivity of chemotherapy, make an analysis of genetic features of ovarian cancer of all periods and predict the recrudescent tendency of ovarian cancer at its early period.The research employs gene expression spectrum to analyze the ovarian serous tumor and normal group .Compare the diversity of the mRNA expression of genes in the two groups, with the hope to filtrate the diversity-expressed genes of the ovarian tumor. Meanwhile, the research also employs the PCR and technique of immunohistochemistry to check respectively in mRNA and protein-level test of gene P13K, hk10 and the expression of prostasin in ovarian tumor tissue, aiming at accumulating experience for finding new marker of ovarian tumor and the diagnosis of early ovarian tumor.Using cDNA microarray to analyze the gene expression spectrum of ovarian tumor and we gets the following findings: 58 diversity-expressed genes, among which up-regulated genes are 30, and down-expressed genes are 28. The genes which play significant role in the generate and development of tumor and ovarian cancer are as follows: AURKA, PTP4A2, PIK3CA, KLK10, PRSS8 (prostasin), and COX-2 are up-regulated genes; and BTG2, BRCA1, CAV1, ST13 are down-expressed genes.hk (KLK 10) is a member of the kallikrein family, which is also called normal epithelial cell-specific-1 gene. One molecular weight is 30KD, including 276 serine proteases of aminophenol secretion. It is combined with some nutrilit, hormone or extracellular matrix and participates in the process of apoptosis,vein genesis and metastatic tumour. Recent years the domestic and the overseas experts study the expression of hk 10 in ovarian cancer tissue and body fluid. They find that hk 10 is significantly high in the serum of ovarian cancer patients, and conclude that hk10 may be a marker of ovarian cancer which more significant than CA125.Phosphatidylinositol 3-Kinase (PI3K), the important signal-transmitting molecules inside lymphocyte and the second lipid messenger related to transduction of cell signal, has the function to improve the cell proliferation and polarization, restrain the apoptosis, vesicle transit, and glucose metabolism. PI3K conducts the signal concerning the growing and survival of cells by various means. PIK3CA, the oncogene, mutates in malignant tumor in human's history. The overseas research reports that PIK3CA may be a new kind of oncogene because it is intensified in the expression of ovarian cancer.Protasin is a kind of trypsinase-like serine proteases which composed of 343 amino acid, and molecular mass is 40kDa. Protasin, highly expressed in prostasin tissues and least expressed in normal testicle and ovarian tissue, is produced mainly in prostate and achieved by distilling sperm. The researcher finds that prostasin expressed more strongly in cancerous epithelium and stroma than in normal ovarian tissue. The prostasin level of the serum of the patients is obviously higher than control group. Joint application with CA125 in the diagnosis of ovarian cancer can achieve sensitivity and specificity, respectively 92% and 94%.Using RT-PCR techniques to check the expression of P13K, hk 10 and prostasin mRNA in ovarian tumor obtains the following findings: (1) P13K mRNA expressed in normal group and ovarian tumor group, and the expression is weaker in ovarian tumor group than that in normal group. There is no obvious difference as for the expression level in the two groups(P>0.05), however, the expression level of P13K mRNA in ovarian cancer group is significantly higher than that in ovarian tumor group and normal group,and there is statistical significance in the results (P<0.01). (2) hk10mRNA shows no obvious difference between ovarian tumor group and normal ovarian(P>0.05), however, the expression amount of hk10 mRNA in ovarian cancer group is significantly higher than that in ovarian tumor group and normal group,and there is statistical significance in the results (P<0.01).(3) Prostasin mRNA expresses in a small amount in the ovarian tissue of the normal group, expresses more in ovarian tumor group, without statistical significance(P>0.05). However, the expression of prostasin in ovarian cancer group is significantly higher than that in the other two groups.The results of examination of PI3K, hk10 and prostasin in ovarian cancer tissue by using technique of immunohistochemistry are: (1) PI3K protein positive expression mainly pitch in cytoplasm and karyon or karyon. PI3K protein has no or weak expression in normal ovarian epithelial cells and ovarian serous tumor epithelial cells, but it has strong expression in ovarian cancer cells and mainly pitches in karyon; the rate of positive expression of PI3K protein in ovarian cancer group is significantly higher than that in normal group and benign tumor group(P<0.01), and the worse the differentiation of the ovarian cancer, the higher the level and the stronger the expression. (2) hk10 protein positive expression part mainly pitches in cytoplasm. Most normal ovarian epithelial cells have no expression, benign ovarian tumor cells have weak expression but ovarian cancer cells have strong expression. The rate of positive expression of ovarian cancer is significantly higher than that in normal group and benign tumor group(P<0.01). And the expression is more significant with the increase of the level of ovarian cancer. (3) Prostasin protein positive expression part mainly pitches in cytoplasm. Normal ovarian epithelial cells and ovarian cyst expresses weak, some cases expresses negative; while ovarian cancer cells express obviously, its intensity of expression is significantly stronger than the previous two groups (P<0.01), the expression intensity of the person with the higher level of ovarian cancer is lower than the person with the higher differentiation.We draw the following conclusions through the research:1.The technique of cDNA microarray can be used to analyze ovarian cancer gene expression spectrum, screen differential expression gene and give a research foundation for further study of gene in the development of ovarian caner.2.In the study of cDNA microarray, PI3K, hk 10, prostasin gene are up– regulated genes.3.PI3K,hk10 and prostasin mRNA have minim expression in ovarian tissues. The expression level of PI3K, hk10 and prostasin mRNA in ovarian serous tumor tissues is close to normal ovarian tissues. But the expression quantity of PI3K, hk10 and prostasin mRNA in ovarian serous bursal cancer tissues is significantly higher than that of in normal ovarian group and ovarian tumor group.4.The expression of PI3K protein, prostasin protein and hk 10 protein is mainly pitch in ovarian epithelial cytoplasm, but PI3K protein in ovarian cancer has significant expression in karyon.5.The expression of PI3K protein, prostasin protein and hk 10 protein in ovarian cancer cells is significantly higher than that of in nomal ovary and benign ovarian tumor tissues, and it has negative correlation with the degree of differentiation of ovarian cancer., that is the worse the differentiation, the stronger the expression.6. Using cDNA microarray to screen the gene of differentiate expression, which can afford new basis about special marker of ovary cancer,which also can afford the new method of ovary cancer early diagnosis. To sum up, the research employs gene expression spectrum to analyze the ovarian serous tumor and compare the diversity of the mRNA expression of genes in the two groups, with the hope to filtrate the diversity-expressed genes of the ovarian tumor. Meanwhile, the research also employs the PCR and technique of immunohistochemistry to check respectively in mRNA and protein-level test of gene P13K, hk10 and the expression of prostasin in ovarian tumor tissue, aiming at accumulating experience for finding new marker of ovarian tumor and the diagnosis of early ovarian tumor. |
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