Application of new tissue microarrayer-ZM-1 without recipient paraffin block

The ZM-1 tissue microarrayer designed by our groups is manufactured in stainless steel and brass and contains many features that make TMA (tissue microarray) paraffin blocks construction faster and more convenient. By means of ZM-1 tissue microarrayer, biopsy needles are used to punch the donor tissue specimens respectively. All the needles with the punched specimen cylinders are arrayed into the array-board, with an array of small holes dug to fit the needles. All the specimen cylinders arraying and the TMA paraffin block shaping are finished in only one step so that the specimen cylinders and the paraffin of the TMA block can very easily be incorporated and the recipient paraffin blocks need not be made in advance, and the paraffin used is the same as that for conventional pathology purpose. ZM-1 tissue microarrayer is easy to be manufactured, does not need any precision location system, and so is much cheaper than the currently used instrument. Our method’s relatively cheap and simple ZM-1 tissue microarrayer technique of constructing TMA paraffin block may facilitate popularization of the TMA technology.

Expression and Significance of Survivin mRNA in Lung Cancer of Different Progression Stages by FISH and Tissue Microarray Technology*

Objective： To investigate the expression of Survivin mRNA in lung cancer progression tissue microarray by FISH （fluorescence in situ hybridization） method and determine its role and significance in lung cancer genesis and progress. Methods： The expression of Survivin mRNA was detected by FISH method and tissue microarray technology. 89 cases of primary lung cancer, 12 cases of lymph node metastasis of lung cancer, 12 cases of precancerous lesion and 10 cases of normal lung tissue were examined. Results： 69.7% of primary lung cancer express Survivin mRNA; the positive ratio of primary lung cancer and precancerous lesion were both significantly higher than that of normal lung tissue （P〈0.05）; the expression of Survivin mRNA was related to the differentiation degree, lymph node metastasis and clinical stages （P〈0.05）. Conclusion： FISH has good sensitivity and stability. Tissue microarray technology has many advantages, such as high efficiency, high throughput, etc; it may have good prospect in pathology. Survivin mRNA was highly expressed in lung cancer and precancerous lesion; it was related to the progress and malignant behavior; it may play a promotion role in lung cancer genesis and progress and offer basis to early diagnosis, prognosis estimate and treatment.

Expression of COX-2 in Different Subtypes of Gastric Intestinal Metaplasia and Gastric Carcinoma by Tissue Microarray

Objective: To study the expression of cyclooxygenase-2 (COX-2) protein in different subtypes of intestinal metaplasia (IM) and gastric carcinoma, evaluate the possibility of COX-2 forecasting the risk of malignant potential of IM, and the relationship between COX-2 expression and gastric carcinogenesis. Methods: Forty cases of chronic atrophic gastritis (CAG) with IM, 40 cases of gastric carcinoma and corresponding paracancerous tissues were selected to construct a tissue microarray. High iron diamine/alcian blue (HID/AB) staining and Hematoxylin and Eosin (HE) staining was used to classify IM and gastric carcinoma, and the expression of COX-2 protein detected in different subtypes of IM and gastric cancer by using immunohistochemistry. Results: The positive expression rate of COX-2 was 45.65%, 59.38% and 77.27% in IM foci in CAG, IM foci in paracancerous tissues, and intestinal-type gastric carcinoma, respectively, significantly higher than in diffuse-type gastric cancer (16.67%)(P<0.05, 0.005 and 0.005, respectively), and the expression intensity of COX-2 protein showed a increased tendency gradually in the sequence of IM foci in CAG→IM foci in paracancerous tissues→intestinal-type gastric carcinoma (P<0.005). The positive expression rate of COX-2 protein in type Ⅲ IM was significantly higher than in type Ⅰ and type Ⅱ IM (P<0.005 and 0.05, respectively), and the expression intensity also showed a increased tendency gradually from type Ⅰ to type Ⅲ IM (P<0.005). Conclusion: The expression level of COX-2 was increased gradually along with the increase of the risk of malignancy of IM, and its expression level may be a useful index to forecast the risk of malignant potential of IM. COX-2 expression was associated with intestinal-type gastric carcinoma, but it might also have some role in the carcinogenesis of diffuse-type gastric carcinoma.

COX-2 expression in gastric cancer and its relationship with angiogenesis using tissue microarray

AIM: To explore the expression and clinicopathological significance of cyclooxygenase-2 (COX-2) and microvessel density (MVD) in gastric carcinogenesis, and to investigate their roles in the invasion and the relationship between biological behaviors and prognosis of gastric cancer. METHODS: Using Envision immunohistochemistry, COX-2 and CD34 expressions in gastric cancer tissue array were examined. MVD was counted and the relationship between the biological behaviors and prognosis was analyzed. RESULTS: The expression of COX-2 in gastric cancer tissue was significantly higher than that in normal mucosa (χ2 = 12.191, P < 0.05). The over-expression of COX-2 in gastric cancer was obviously related to metastasis and depth of invasion (χ2 = 6.315, P < 0.05), but not related to the histological type and Borrmann type (χ2 = 5.391 and χ2 = 2.228, respectively). Moreover, MVD in gastric cancer tissues was significantly higher than that in the normal mucosa (65.49 ± 20.64 vs 36.21 ± 18.47, t/F = 7.53, P < 0. 05). MVD was related to the histologic type and metastasis (t/F = 3.68 and t/F = 4.214, respectively, P < 0. 05), but not related to the depth of invasion and Borrmann type (t/F = 0.583 and t/F = 0.459, respectively). MVD in COX-2-positive tissues was markedly higher compared to COX-2-negative tissues, indicating a positive correlation between COX-2 expression and MVD (t = 13.12, P < 0. 05). CONCLUSION: Tissue microarray (TMA) is a powerful tool for rapid identifi cation of the molecular alterations in gastric cancer. COX-2 expression, via inducingangiogenesis, may play an important role in gastric carcinogenesis. It could be served as a determinant factor for clinical prognosis and curative effect.

Tissue microarray for high-throughput analysis of gene expression profiles in hepatocellular carcinoma

AIM: To study the expression profiles of HBsAg, HBcAg,p21WAF1/CIP1 (p21), Rb genes in hepatocellular carcinoma (HCC) and to investigate their roles in the hepatocarcinogenesis.METHODS: HCC tissue microarray containing 120-min tissues of 40 HCC cases was constructed. HBsAg, HBc Ag,p21 and Rb proteins were immunohistochemically stained by streptavidin-peroxidase conjugated method (S-P). The expression loss of these genes in cancerous, paracancerous tissues and adjacent normal liver tissues of 40HCCs were comparatively examined.RESULTS: The positive rate of HBsAg expression in cancerous tissues of 40 HCCs was 7.5%, which was lower than that in para-cancerous and adjacent normal liver tissues (x2 =12.774, P＜0.01; x2= 18.442, P＜0.01). The positive rate of HBcAg expression in cancerous tissues of 40 HCCs was 20.0%, which was also lower than that in para-cancerous and adjacent normal liver tissues(x2= 9.482, P＜0.01; x2= 14.645, P＜0.01). p21 protein deletion rate in cancerous tissues of 40 HCCs was 27.5%,which was higher than that in para-cancerous and adjacent normal liver tissues (x2 = 7.439, P＜0.01; x2 = 11.174,P＜0.01). p21 protein deletion correlated remarkably with the pathological grade of HCC (x2 = 0.072, P＜0.05). Rb protein deletion rate in cancerous tissues of 40 HCCs was42.5%, which was also higher than that in para-cancerous and adjacent normal liver tissues (x2 = 10.551, P＜0.01;x2= 18.353, P＜0.01). Rb protein deletion rate did not correlate remarkably with tumor size or pathological grade of HCC (x2 = 0.014, P＞0.05; x2 = 0.017, P＞0.05).CONCLUSION: Expression deletion of HBsAg, HBcAg, p21and Rb proteins in HCCs may play important roles in the carcinogenesis of HCC. Tissue microarray is an effective high-throughput technique platform for cancer research.

Tissue array for Tp53,C-myc,CCND1 gene over-expression in different tumors

AIM:To rapidly detect molecular alterations in different malignancies and investigate the possible role of Tp53,C-myc,and CCND1 genes in development of tumors in human organs and their adjacent normal tissues,as well as the possible relation between well-and poorly-differentiated tumors.METHODS:A tissue array consisting of seven different tumors was generated.The tissue array included 120 points of esophagus,120 points of stomach,80 points of rectum,60 points of thyroid gland,100 points of mammary gland,80 points of liver,and 80 points of colon.Expressions of Tp53,C-myc,and CCND1 were determined by RNA in situ hybridization.3' terminal digoxin-labeled anti-sense single stranded oligonucleotide and locked nucleic acid modifying probe were used.RESULTS:The expression level of Tp53 gene was higher in six different carcinoma tissue samples than in paracancerous tissue samples with the exception in colon carcinoma tissue samples(P < 0.05).The expression level of CCND1 gene was significantly different in different carcinoma tissue samples with the exception in esophagus and colon carcinoma tissue samples.The expression level of C-myc gene was different in esophagus carcinoma tissue samples(c2 = 18.495,P = 0.000),stomach carcinoma tissue samples(c2 = 23.750,P = 0.000),and thyroid gland tissue samples(c2 = 10.999,P = 0.004).The intensity of signals was also different in different carcinoma tissue samples and paracancerous tissue samples.CONCLUSION:Over-expression of the Tp53,CCND1,and C-myc genes appears to play a role in development of human cancer by regulating the expression of mRNA.Tp53,CCND1 and C-myc genes are significantly correlated with the development of different carcinomas.

Expression of p53, p16 and COX-2 in pancreatic cancer with tissue microarray

BACKGROUND: Pancreatic cancer development and progression is driven by the accumulation of genetic changes. In this study we constructed tissue microarray containing specimens from pancreatic cancer, adjacent non-cancer tissue and normal tissue to survey the expression of p53, p16 and cyclooxyganase-2 (COX-2). METHODS: Tissue microarray containing 337 specimens from different stages of pancreatic cancer, adjacent noncancer tissue and normal tissues was constructed, and the expression of p53, p16 and COX-2 was assayed by immunohistochemistry to consecutive formalin-fixed tissue microarray sections. RESULTS: The expression of p53, p16 and COX-2 was significantly higher in tumorous tissues than in non-tumorons ones. A significant relationship was observed between p53 and COX-2, or p16 and COX-2. But no obvious correlation was seen between p53 and p16 expressions. Logistic regression analysis showed p53 and COX-2 as dependent predictors in pancreatic carcinogenesis, and a reciprocal relationship to neoplastic progression between p53 and COX-2. CONCLUSION: Combination analysis of p53 and COX-2 may be useful in predicting pancreatic carcinogenesis.

EXPRESSION AND SIGNIFICANCE OF SURVIVIN mRNA IN LUNG CANCER TISSUE MICROARRAY DETECTED BY FISH

Objective To investigate the expression of Survivin mRNA in lung cancer tissue microarray （TMA） by fluorescence in .situ hybridization （FISH） method, and determine the role and significance of it in lung cancer genesis and progress. Methods The expression of Survivin mRNA was detected by FISH method and TMA technology. Fifty-four cases of lung cancer and 10 cases of normal lung tissue were examined. Survivin mRNA was expressed in 66.7% （36/54） of lung cancer; the positive ratio of lung cancer was significantly higher than that of normal lung tissue （0/10;X^2= 15.238, P 〈 0.05）. The positive ratio of Survivin mRNA was significantly higher in poor differentiated cancer （20/24, 83.3% ） than moderate and well differentiated cancer （16/30, 53.3%; X^2 = 5.40, P 〈 0.05）. The positive ratio of Survivin mRNA was significantly higher in group with lymph node metastasis （27/32, 84.4%） than without lymph node metastasis （9/22, 40.9%; X^2= 11.084, P 〈 0.05）. The positive ratio of Survivin mRNA was significantly higher in stage Ⅲ-Ⅳ（12/13, 92.3%） than stage Ⅰ- Ⅱ （24/41,58.5%; X^2=5.066, P〈 0.05）. Conclusion Survivin mRNA highly expresses in lung cancer, which is related to the progress and malignant behavior. Survivin may play a promoting role in lung cancer genesis and progress and provide a basis for estimating prognosis and treatment.

EXPRESSION OF SURVIVIN,PTEN AND BFGF IN LUNG CANCER PROGRESSION TISSUE MICROARRAY

To investigate the expressions of survivin, PTEN and bFGF in lung cancer and precancerous lesion by tissue microarray technology. Methods: The expressions of Survivin, PTEN and bFGF were detected in 89 primary lung cancers, 12 lung cancers with lymph node metastasis, 12 precancerous lesions of lung by an immunohistochemical method, and 10 normal lung tissues were used as controls. Results: The expression of Survivin and bFGF protein were 57.3% and 66.3% in 89 primary lung cancer respectively, significantly higher than the control group (P<0.05). The positive ratio of PTEN was 42.7% in primary lung cancer, significantly lower than that of nornmal tissue (90.0%)(P<0.05). The expressions of Survivin and bFGF were significantly related to lymph node metastasis and clinical stages. The expression of PTEN was related to differentiations, lymph node metastasis and clinical stages (P<0.05). There was a negative correlation between expressions of Survivin and PTEN, a positive correlation between expressions of Survivin and bGFG (P<0.01). Conclusion: Survivin, PTEN and bFGF proteins may be related to the pathogenesis, progression and malignant behavious of lung cancer, and there are some relationships between expressions of Survivin, PTEN and bFGF. Survivin, PTEN and bFGF may play a role in prognosis assessment of lung cancer.

Significance and expression of Bax, Survivin and p53 in gastric carcinoma and precancerous lesions using tissue microarray

Objective： To explore the relationship between expressions of apoptosis-related protein Bax, Survivin and p53 and the molecular mechanisms of carcinogenesis and progression of gastric carcinoma. Methods： Tissue microarray and immunohistochemistry were used in this study. Results： The positive rate of Bax protein in gastric cancer （17.7%, 17/96） was significantly lower than those in adjacent normal mucosa （51%）, intestinal metaplasia （69.2%） and dysplasia （75%）, P 〈 0.01. The positive rate of Survivin expression in gastric cancer （80.6%, 89/98） was significantly higher than that in adjacent normal mucosa （3.9%）, P 〈 0.01. The positive rates of Survivin expression in tumors with different organ metastases （in lymph node metastasis 86.2%, liver 100% and ovarian 100%） were statistically higher than in tumors without metastasis （64.3%）, P 〈 0.05. Bax expression was correlated with Survivin but not with rap53 that was closely related to Survivin expression （P 〈 0.05） in gastric cancer. Conclusion： The abnormal expressions of Bax, Survivin and rap53 were correlated with the tumorigenesis and progression of gastric carcinoma. P53 and Survivin genes may share the similar mechanism in regulating cell apoptosis, and because of the mutation, p53 gene may lower its down-regulation to Survivin expression.

STAT1 and Survivin Expression in Full Lymph Node Examined Gastric Cancer by Using Tissue Microarray Technique

Objective： To characterize the relationship between STAT1 and Survivin expression, and the relationship between them and lymph node metastasis, depth of invasion and prognosis in full lymph node examined gastric cancer patients of China. Methods： Specimens of curative dissection between 1988 and 2003 were collected from the affiliated hospital of Jianghan University. All 140 patients had complete examination data. All lymph nodes were found by clearing fat method. The interrupted serial 4 μm sections, routine hematoxylin and eosin staining and immunohistochemical methods were used to detect the lymph node metastases. Gastric cancer tissue microarray was formed and the expression of survivin and STAT1 in gastric cancer was detected by immunohistochemical method. All data were processed using Spearman rank correlation analysis, Kaplan-Meyer Log-rank method and Cox multivariate analysis （SPSS 12.0 software）. Results： Among 140 gastric cancer tissue microarrays constructed, 110 could be used （utilization rate was 78.6%）. 7079 lymph nodes were found in 110 cases （64.4/case）. Metastases were found in 89 cases and 1679 lymph nodes. Positive expression rate of survivin and STAT1 was 52.7% （58/110） and 40% （44/110） respectively. There was a significant negative correlation between STAT1 expression and survivin expression （r=-0.19, P=0.04）. STAT1 expression had a negative correlation with depth of invasion （r=0.21, P=0.04）. Survivin expression had a negative correlation with UICC N stage （r=-0.24, P=0.01） and histological classification （r=-0.21, P=0.03） by Spearman rank correlation analysis. But survivin and STAT1 expression was not related with prognosis. A significant correlation between lymph node metastasis and prognosis was demonstrated by Cox multivariate analysis （X^2=4.85, P=0.028）. Conclusion： STAT1 has a negative correlation with survivin expression in gastric cancer. Both of them have no correlation with prognosis in gastric cancer. STAT1 expression can be a molecular marker to predict advanced gastric cancer and survivin a molecular marker of lymph node metastasis in gastric cancer. UICC N stage is the most important prognostic factor in gastric caner in China.

Assessment of XAF1 as A Biomarker to Differentiate Hepatocellular Carcinoma from Nonneoplastic Liver Tissues

Objective： XIAP-associated factor 1 （XAF1） expression has been shown to be related with apoptosis in hepatocellular carcinoma （HCC）. However, the correlation of XAF1 expression with HCC tumor grade has not been intensively assessed. XIAP-associated factor-1 （XAF1） is an important apoptosis inducer in human HCC. The aim of this study is to determine the correlation between XAF1 expression and HCC histopathological grades. Methods： The mRNA levels of XAF1 in 24 paired HCC-nonneoplastic specimens were quantified by real-time reverse transcription PCR （RT-PCR）. Protein levels of XAF1 in 110 paired HCC-noncancer tissues were investigated by immunostaining specimens on a tissue microarray （TMA）. Correlations between XAF1 mRNA levels or protein expression and clinicopathological features were assessed by statistical analysis. Results： Both XAF1 mRNA and protein were significantly under-expressed in HCC tissues compared to their non-neoplastic counterparts. No significant relationship was found between XAF1 mRNA or protein expression and histological tumor grade. Conclusion： All these data suggest that XAF1 is a potential biomarker for differentiating HCC with noncancerous tissues.

ADVANTAGES AND APPLICATIONS OF TISSUE MICROARRAY TECHNOLOGY ON CANCER RESEARCH

S To provide evidences for exploiting tissue microarray (TMA) technology, we reviewed advantages and applications of TMA on tumor research. TMA has many advantages, including (1) section from TMA blocks can be utilized for the simultaneous analysis of up to 1,000 different tumors at DNA, RNA or protein level; (2) TMA is highly representative of their donor tissues; (3) TMA can improve conservation of tissue resources and experimental reagents, improve internal experimental control, and increase sample numbers per experiment, and can be used for large-scale, massively parallel in situ analysis; (4) TMA facilitates rapid translation of molecular discoveries to clinical applications. TMA has been applied to tumor research, such as glioma, breast tumor, lung cancer and so on. The development of novel biochip technologies has opened up new possibilities for the high-throughput molecular profiling of human tumors. Novel molecular markers emerging from high-throughput expression surveys could be analyzed on tumor TMA. It is anticipated that TMA, a new member of biochip, will soon become a widely used tool for all types of tissue-based research. TMA will lead to a significant acceleration of the transition of basic research findings into clinical applications.

Reliability of a Tissue Microarray in Detecting Thyroid Transcription Factor-1 Protein in Lung Carcinomas

OBJECTIVE To compare the expression of the thyroid transcription factor-1 （TTF-1） in human normal adult type Ⅱ alveolar epithelial cells, embryonic pneumocytes and cancer cells of lung carcinoma and metastatic lymph nodes using a tissue microarray （TMA） along with paired conventional full sections, and to investigate the reliability of tissue microarrays in detecting protein expression in lung carcinoma. METHODS A lung carcinoma TMA including 765 cores was constructed. TTF-1 protein expression in both TMA and paired conventional full sections were detected by the immunohistochemical SP method using a monoclonal antibody to TTF-1. A PU （Positive Unit） of TTF-1 protein was assessed quantitatively by the Leica Q500MC image analysis system with results from the paired conventional full sections as controls. RESULTS There was no significance between TMA and paired conventional full sections in TTF-1 expression in different nuclei of the lung tissue. CONCLUSION TTF-1 protein expression in lung carcinoma detected by TMA was highly concordant with that of paired full sections. TMA is a reliable method in detecting protein expression.

Biological Significance and the Related Molecular Mechanism of Ets1 mRNA Expression in Lung Cancer by Tissue Microarray(TMA)

Objective： To investigate the expressions and proteins in the pathogenesis, progression of lung molecular mechanism of Ets-1 mRNA, and TGFβ1 and c-Met cancer by tissue microarray （TMA） method. Methods： The expressions of Ets-1 mRNA, and TGFβ1 and c-Met proteins were detected in 89 primary lung cancers, 12 lung cancer with lymph-node metastasis and 12 precancerous lesions by FISH（fluorescence in situ hybridization） and immunohistochemical method, and 10 normal lung tissues were used as controls. Results： The expressions of Ets-1 rnRNA, and TGFβ1 and c-Met proteins were significantly higher in 89 primary lung cancer than in the control group （P〈0.05）. The expressions of Ets-1 mRNA, and TGFβ1 and c-Met proteins were related to lymph node metastasis and clinical stages. There was a positive correlation between the Ets-1 mRNA expression and TGFβ1 and c-Met proteins （P〈0.05）. Conclusion： Ets-1 mRNA, TGFβ1 and c-Met proteins may be related to the pathogenesis, progression and malignant behavior of lung cancer. They may play an important role in prognosis assessment of lung cancer.

Expression of Elk-1 in Non-Small Cell Lung Cancer Detected by Western Blot and Tissue Microarray

Objective： The aim of this study was to investigate the Elk-1 （Ets like transcription factor-1） expression in non-small cell lung cancer （NSCLC） and normal lung tissues and the relationship between its expression and clinicopathological characters. Methods： To observe Elk-1 expression, western blot and immunochemistry （IHC） on tissue microarray （TMA） containing 118 lung cancers and their corresponding normal tissues were used. Results： In western blot and IHC on TMA, Elk-1 was highly expressed in NSCLC, while its expression was almost undetectable in normal lung tissues. Elk- 1 expression in NSCLC had no relationship with the patients＇ age, gender, smoking status and histological type, but had relationship with the differentiation degree, clinical stages and lymphonode metastasis. The expression was lower in early stage group （Ⅰ＋Ⅱ） than in advanced stage group （Ⅲ）, and lower in well-moderately differentiated group than in poorly differentiated group. The same trend was seen with lymphonode metastasis. Conclusion： The progression of NSCLC may be related with the increased Elk-1 expression, and Elk-1 may be regarded as a prognostic factor for NSCLC tissues.

Identification of p63 expression in human lung cancer: analysis by complementary DNA and tissue microarray

Objective: To evaluate p63 expression at mRNA transcripts and protein levels in lung squamous cell cancer (SCC), adenocarcinoma, large cell lung cancer (LCLC) and small cell lung cancer (SCLC) and their matched metastatic tumors. The association between p63 expression and p63 locus at chromosomal 3q27 q29 was also investigated. Methods: p63 mRNA expression levels in a large series of lung cancers including SCC, adenocarcinoma, LCLC, SCLC and their matched metastatic tumors were analyzed by cDNA microarray technology. A tissue microarray from 150 primary lung cancer specimens was constructed and used for immunohistochemical detection of p63 protein expression. Chromosomal imbalances at the p63 locus in 70 primary lung cancers samples were studied by comparative genomic hybridization (CGH) technology. Results: mRNA levels were 10 fold in SCC compared to LCLC, SCLC, and adenocarcinoma. Interestingly, the mRNA expression of p63 in metastatic carcinomas was significantly higher than that in their matched primary tumors ( P <0 001). Immunohistochemistry demonstrated that p63 expression was 94.64% in SCC but only 1 79% in lung adenocarcinoma and 2 of 4 LCLC were positive staining. All the results in of SCLC were negative. There was a statistically significant difference for p63 positivity between pT1 tumors and those of higher stage ( P =0 035). The CGH results indicated that p63 locus at chromosomal 3q27 q29 was overrepresented in SCC. p63 immunopositivity correlated significantly with pronounced gains of the p63 locus at chromosomal 3q27 q29 (P=0.0001), indicating that strong expression of p63 in lung SCC correlated with increased gene amplification. Conclusion: p63 might play an important role not only in squamous differentiation of lung cancer but also in tumor development and progression.

Evaluate the Expression of uPA,PAI-1 in Human Gastric Cancer and its Correlation with the Angiogenesis by the Application of Tissue Microarray

OBJECTIVE To investigate the expression of urokinase-type plasminogen （uPA）, its inhibitor-1 （PAI-1） mRNA and its protein in human gastric cancer and to find out the relationship among the tumor differentiation, angiogenesis, and other clinical pathologic factors. METHODS In situ hybridization （ISH） was used to get the uPA, PAI-lmRNA in 110 cases with human gastric cancer in 2-tissue microarray （TMA）. Immunohistochemical staining （S-P method） for uPA, PAI-1 protein and CD34 were performed in the 110 cases in 2 TMA. RESULTS The expression of the uPA, PAI-lmRNA and their protein happened in the cytoplasm of gastric cancer cells were induced by the poor differentiation of the GC, and the expression of uPA had an increasing trend while the expression of the PAI-1 had a decreasing trend. The microvessel density （MVD） had a positive correlation with the clinical stages and the significant relationship with the lymph node metastasis （P 〈 0.05）. The MVD in uPA positive group was significantly higher than those in uPA negative group （P 〈 0.05）. The expression of PAI-1 has no correlation neither with the clinical stages nor the lymph node metastasis. CONCLUSION The uPA play an important role in invasion and metastasis of GC through promoting angiogenesis. Interdicting the secretion and function of the uPA may allow the target therapy against the tumor invasion. As a new high-throughput technology, the tissue microarray is a valuable way to be used in clinical treatment.

Construction of metastatic spinal cancer tissue microarrays

Objective： To explore the construction of metastatic spinal cancer （MSC） tissue microarrays and validate its value in immunohistochemical study of MSC. Methods： Paraffin-embedded specimens from 71 MSC cases and 6 primary tumor cases were selected as donor blocks and prepared into MSC tissue microarrays by tissue array arrangement, the steps of which included location, punching, sampling, sample seeding, and re-diagnosis by hematoxylin-eosin （HE） as well as MMP-9 and MMP-14 immunohistoehemical staining. Results： The MSC tissue microarrays thus constructed were intact and craekless, containing 154 complete and well arranged microarray points. None of the sectioned tissue microarrays was lost, and the results of HE staining was consistent with the primary pathologic diagnoses. Immunohistochemical staining was also good without non-specific or marginal effect. Conclusion： The MSC tissue microarrays have a high value in the immunohistochemical study of MSC.

Study on the Expression and Significance of TGF-β1, p-ERK1/2and K-ras in Colorectal Cancer Using Tissue Microarray Technique

OBJECTIVE This study aimed to explore the expression and significance of transforming growth factor β1（TGF-β1）,extracellular signal-regulated kinases 1/2 （ERK1/2）, and K-ras in colorectal cancer （CRC） using tissue microarray technology.METHODS The expressions of TGF-β1, ERK1/2, and K-ras in colon cancer cells taken from the specimens of 92 CRC patients （stage Ⅰ： 16 cases, stage Ⅱ： 28 cases, stage Ⅲ： 24 cases, and stage Ⅳ：24 cases） were analyzed using tissue microarray technology and immunohistochemistry, and compared with those of 20 normal colon tissue samples.RESULTS High immunoreactive scores （IRS） of TGF-β1,p-ERK1/2, and K-ras protein in CRC were obtained, which were 66.3% （61/92）, 59.8% （55/92）, and 48.9% （45/92）, respectively, and those in normal epithelial cells of colon were 10% （2/20）, 20% （4/20）, and 30% （6/20）, respectively （P 〈 0.05）. The expressions of TGF-β1 and ERK1/2 in CRC at stage Ⅰwere 37.5% and 31.3%,respectively, and those in CRC at stage Ⅳ were 83.3% and79.3%, respectively, with statistically significant differences. No significant relationship was found between K-ras expression and tumor stages （P〉0.05）.CONCLUSION High level expressions of TGF-β1 and ERK1/2 are closely related to the clinical stages of colon cancer and crosstalk may exist between the 2 signal pathways.