Expression and Clinical Significance of REGy in Gastric Cancer Tissue and Variously Differentiated Gastric Cancer Cell Lines

OBJECTIVE To evaluate the REGy expression in gastric cancer tissue and gastric cancer cell lines of various differentiation levels and its clinical significance.METHODS Immunohistochemistry was used to detect the expression of REGy protein in 70 specimens of gastric cancer and 30 specimens of normal gastric mucosa. The relationship between the expression of REGy protein and the biological behaviors of gastric cancer was analyzed. RT-PCR and Western blot were used to detect the mRNA level and the protein expression of REGγ in normal gastric cell line GES-1, well differentiated gastric cancer cell line MKN-28, moderately differentiated gastric cancer cell line SGC-7901 and poorly differentiated gastric cancer cell line BGC-823.RESULTS The expression rate of REGγprotein in gastric cancer tissue (52/70, 74.29%) was significantly higher than that in normal gastric tissue (12/30, 40%) (P<0.01). The expression rate of REGywas correlated with tumor size (P<0.01), lymph node metastasis (P<0.05), differentiation degree (P<0.01), infiltration depth (P<0.01)and distant metastasis (P<0.05). RT-PCR analysis showed that theexpression of REGγ mRNA was 0.459±0.079 in the normal gastric mucosa cell line, 0.588±0.118 in the well differentiated gastric cancer cell line, 0.715±0.066 in the moderately differentiated gastric cancer cell line, and 0.873±0.099 in the poorly differentiated gastric cancer cell line, showing a negative correla- tion between REGγmRNA expression and differentiation level (P <0.05). Western blot analysis showed that the expression of REGy protein was 0.712±0.065 in the normal gastric mucosa cell line, 1.176±0.185 in the well differentiated gastric cancer cell line, 1.533 ±0.127 in the moderately differentiated gastric cancer cell line, and 2.061±0.398 in the poorly differentiated gastric cancer cell line, showing a negative correlation between REGγprotein expression and differentiation level (P<0.05).CONCLUSION REGγ is expressed in gastric cancer tissue and normal gastric tissue. In gastric cancer tissues, REGγexpression is positively correlated with the tumor size, lymph node metastasis,differentiation degree, infiltration depth and distant metastasis. Detecting the expression of REGy mRNA and protein is helpful for early diagnosis and predicting prognosis of gastric cancer.     

Significance of the Expression of Rho-GDP Dissociation Inhibitorβ,γin Lung Squamous Cell Carcinoma and Adenocarcinoma

Objective:To study the expression of Rho-GDP dissociation inhibitor β,γ(Rho-GDIβ,Rho-GDIγ)in lung squamous cell carcinoma and adenocarcinoma and its relationship with the expression of RhoC(Ras homologus oncogenes C)and clinicopathologic parameters.Methods:Western blot assay was employed for Rho-GDIβ,Rho-GDIγ and RhoC in lung squamous cell carcinoma and adenocarcinoma and non-neoplastic lung tissues of 37 cases with fresh specimens.Results:The study showed that Rho-GDIβ,Rho-GDIγ and RhoC were expressed in lung cancer and non-neoplastic lung tissues,the level in lung cancer tissue was much higher than that in non-neoplastic tissues(P<0.001).In lung cancer,the expression of Rho-GDIβ was much higher in patients with lymph node metastasis(P=0.021),and the expression of Rho-GDIγ was much higher in poorly differentiated tumor than in well-differentiated and moderately differentiated tumor,but both of them were not correlated with other clinicopathologic parameters.The expressions of Rho-GDIβ and Rho-GDIγ were not correlated with the expression of RhoC.Conclusion:In lung cancer,Rho-GDIβ and Rho-GDIγ may play a role in the tumorigenesis,Rho-GDIβ may promote metastasis,and Rho-GDIγ may have some relationship with differentiation.     

DETECTING EXPRESSION OF MRP-1/CD9 mRNA IN LUNG CANCERS USING TISSUE MICROARRAYS AND FLUORESCENCE IN SITU HYBRIDIZATION METHODS

Objective： The aim of this study was to investigate the MRP-1/CD9mRNA expression in lung cancer and normal lung tissues and the relationship between its expression and pathologic grades, clinical stages, metastasis and prognosis. Methods： To observe MRP-1/C9mRNA expression, tissue microarray （TMA） containing 54 lung cancers and 10 normal lung tissues was prepared and Fluorescence in situ hybridization was used. Results： The positive rate of MRP-1/CD9 expression was 48.1% in lung cancer, lower than that of normal lung tissues. The statistical difference was significant （P〈0.05）. Its protein expression had no relationship with the patients＇ ages, sex and the macroscopic type of tumor, but had relationships with the histological type, clinical stage, differentiated degree and metastasis. The expression in non-small cell lung cancer （NSCLC） was higher than that in small cell lung cancer （SCLC）; in well-moderately differentiated group was higher than that in poorly differentiated group; Earlier period group （I＋II） was higher than in later period group （Ⅲ＋Ⅳ）; and in group without lymphoid metastasis was higher than in patients with lymphoid metastasis. Conclusion： The progression of the lung cancer maybe related with the descended MRP-1/Cd9 expression, which may be useful in evaluating the prognosis of cancer patients.     

The Effect of Helicobacter pylori Infection on hMSH2 and P53 Proteins in Gastric Carcinogenesis

OBJECTIVE To investigate the effect of Helicobacter pylori （H pylon） infection on expression of hMSH2 and P53 and its possible carcinogenic mechanism. METHODS H pylori infection was detected using a rapid urease test and haematoxylin and eosin （H＆E） staining. The hMSH2 and P53 proteins in gastric cancer （GC） tissue, its adjacent mucosa, gastritic mucosa and normal gastric mucosa were detected by the immunohistochemical SP method. RESULTS （1） The positive rate of hMSH2 expression in GC tissue （62.7%）was higher than those in non-cancer tissues （P〈0.001）; the positive rate of hMSH2 expression in poorly differentiated adenocarcinoma （76.4%） was higher than that in other carcinomas （P〈0.05）. The positive rate of P53 expression in GC tissue （51.9%） was higher than that in noncancer tissues （P〈0.001）; the positive rate of P53 expression in well-differentiated adenocarcinoma （32.6 %） was lower than that in other carcinomas （P〈0.01）. （2） The positive rate of hMSH2 expression in GC tissue with H pylori infection was lower than that without the infection （52.8% vs. 74.5%; P〈0.05）. The positive rate of P53 expression in GC tissue with H pylori infection was higher than that without the infection （61.4% vs. 40.6%; P〈0.05）.（3） The expression of hMSH2 and P53 in GC tissue correlated positively （r=0.457, P〈0.01）. CONCLUSION High expression of hMSH2 and P53 as well as their interaction may be involved in gastric carcinogenesis; H pylori infection affecting expression of hMSH2 and P53 may be one of its carcinogenic mechanisms.     

血管内皮生长因子-D在胃癌中表达的意义

Objective： To investigate the expression of vascular endothelial growth factor D （VEGF-D） in gastric cancer and its relationship with lymph node metastasis. Methods： 100 cases of gastric carcinoma tissues （50 cases with lymph node metastasis, 50 cases negative） and 30 cases of normal gastric tissues were gathered to detect the expressions of VEGF-C and D proteins by immunohistochemistry. Results： VEGF-C and D were revealed in cytoplasm of gastric carcinoma tissues and normal gastric tissues. The positive rates of VEGF-C and D expressions were significantly higher in gastric cancer tissues than those in the normal ones （51%, 60% vs 10%, 20% respectively; both P 〈 0.05）. There were significant correlations between the positive expression of VEGF-D and lymph node metastasis, lymphatic invasion, positive expression of VEG F-C, but not with tumour size, tissue differentiation, and venous invasion. Conclusion： The expression of VEGF-D is closely related to lymph node metastasis in gastric carcinoma.     

EXPRESSION OF FRAGILE HISTIDINE TRIAD AND P53 IN NON-SMALL CELL LUNG CANCER

Objective： To investigate the expression offragile histidine triad （FHIT） and p53 protein in non-small cell lung cancer （NSCLC） and explore the relationship between their expressions and the clinicopathological features. Methods： FHIT protein and p53 protein were detected by immunohistochemistry in 76 cases of NSCLCs and matched normal lung tissues. Results： Fifty-one cases （67.1%） showed negative expression of FHIT （apparent reduction or loss） and thirty-seven cases （48.7%） showed p53 positive expression （overexpression）. The difference was significant （P=0.04）. However, there was no significant difference in FHIT expression between the p53-positive group and the p53-negative group （64.9% versus 69.2%, P=0.686）. The negative rate of FHIT protein expression was higher in squamous cell carcinoma than in adenocarcinoma, in moderately and poorly differentiated carcinoma than in well-differentiated carcinoma, and in cases with smoking history than in cases without smoking history （P〈0.05）. There was no relationship between FHIT expression and clinical stage or lymph node metastasis. The negative FHIT expression was not an independent predictor of overall survival （P=0.338）. Conclusion： The frequency of negative expression of FHIT protein is higher than that of positive expression of p53 in NSCLCs. The negative expression of FHIT is independent of the expression of p53. The change of expression of FHIT may play a role in the smoking related lung tumorigenesis while it may have no relationship with the progress of NSCLC or prognosis of the patients.     

利用组织芯片技术研究胃癌及其癌前病变中Bax、p53、Survivin表达的关系及意义

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.     

Level of circulating PD-L1 expression in patients with advanced gastric cancer and its clinical implications简

Objective：The programmed cell death-1 receptor/programmed cell death-1 ligand （PD-1/PD-L1） pathway plays a crucial role in tumor evasion from host immunity.This study was designed to evaluate the association between circulating PD-L1 expression and prognosis in patients with advanced gastric cancer.Methods：Totally 80 advanced gastric cancer patients and 40 health controls from Beijing Cancer Hospital were enrolled in the present study.Circulating PD-L1 expression was tested by enzymelinked immunosorbent assay （ELISA）.The associations between the expression level of PD-L1 and clinicopathological features and prognosis were analyzed statistically.Results：Expression of PD-L1 in advanced gastric cancer patients was significandy up-regulated compared with health people （P=0.006）.The expression of PD-L1 was significantly correlated with differentiation and lymph node metastasis （P=0.026 and P=0.041,respectively）.Although we didn＇t find significant difference in all advanced gastric cancer patients with different PD-L1 expression,the adenocarcinoma patients with higher up-regulated PD-L1 expression had much better prognosis than low expression patients （65.6％ vs.44.7％,P=0.028）.Conclusions：PD-L1 was elevated in advance gastric cancer patients and may play an important role in tumor immune evasion and patients prognosis.     

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.     

Expression of CD44s mRNA in Gastric Carcinoma

Objective： To study the expression of CD44s mRNA in the occurrence, development and invasion of gastric carcinoma （GC）. Methods： The expressions of CD44s mRNA in 66 cases of GC, 25 cases of superficial gastritis and 25 cases of atypical hyperplasia were examined by in situ hybridization （ISH）. Results： There was no expression of CD44s mRNA in the group of superficial gastritis; the positive rate was 20%（5/25） in the group of atypical hyperplasia and 62.12%（41/66） in the group of gastric carcinoma. The positive rate in poor differentiation group was significantly higher than that in well differentiation group （P〈0.05）, and the positive rate of lymph node metastasis group was significantly higher than that in negative lymph node metastasis group（P〈0.05）. Conclusion： The expression of CD44s mRNA was related to cell differentiation degree and lymph node metastasis, the activation of CD44s gene was related to strong invasion of cancer cells and poor prognosis.     

FISH结合组织芯片技术检测不同进展阶段肺癌组织中Survivin mRNA的表达和意义

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.     

Down-regulated expressions of PPARγ and its coactivator PGC-1 are related to gastric carcinogenesis and Lauren＇s classification in gastric carcinoma

Objective： To explore the relationship between peroxisome proliferator activated receptor-gamma （PPARγ） and peroxisome proliferator-activated receptor-gamma coactivator-1 （PGC-1） expression in gastric carcinoma （GC）, and analyze their correlations with clinicopathological features and clinical outcomes of patients. Methods：The two-step immunohistochemical method was used to detect the expression of PPARγ and PGC-1 in 179 cases of GC, and 108 cases of matched normal gastric mucosa. Besides, 16 cases of fresh GC specimens and corresponding normal gastric mucosa were detected for PGC-1 expression with Western blotting. Results： The positive rates of PPART and PGC-1 expression were significantly lower in GC （54.75%, 49.16%） than in normal gastric mucosa （70.37%, 71.30%）, respectively （P〈0.05）. The decreased expression of PGC-1 in GC was confirmed ha our Western blot analysis （P=0.004）. PPAR7 and PGC-1 expressions were related to Lauren＇s types ofGC （P〈0.05）. Positive correlation was found between PPART and PGC-1 expression in GC （rk=0.422, P〈0.001）. The survival time of PPART negative and positive patients was 36.6±3.0 vs. 38.5_＋2.7 months, and no statistical difference was found between the 5-year survival rates of two groups （34.4% vs. 44.1%, P=0.522, log-rank test）; the survival time of PGC-1 negative and positive patients was 36.2±2.8 vs. 39.9±2.9 months, while no statistical difference was found between the 5-year survival rates of the two groups （32.0% vs. 48.2%, P=0.462, log-rank test） Conclusions＇. Decreased expression of PPARγand PGC-1 in GC was related to the Lauren＇s classification. Their expressions in GC were positively correlated, indicating that their fimctions in gastric carcinogenesis may be closely related.     

KiSS-1和S100A4在胃癌中的表达变化以及与转移的关系

Objective： To detect the expression of KISS-1 and S100A4 in the primary tumor tissues and lymphatic and visceral metastases and investigate its role in tumorigenesis and metastasis of gastric cancer. Methods： The protein expression of KISS-1 and S100A4 in lymphatic and visceral metastases from advanced gastric cancer specimens was mainly examined by immunohistochemical staining and tissues microarray. Results： Immunohistochemical staining revealed reduced expression of KISS-1 and up-regulated expression of S100A4 in lymph node and visceral metastases. Rates of KISS-1 expression in normal tissues, primary tumor tissues, lymph node and visceral metastases were 95.8%, 74.6%, 60.9% and 57.5%. $100A4 expression in associated cases was 43.6%, 71.8%, 70.3% and 90.0%, respectively. Significant differences in KISS-1 expression was significantly higher in normal tissues than that in primary tumor tissues （P〈0.001）. While significant differences of S100A4 expression could be seen between normal and cancer tissues （P〈0.001） and between visceral and primary tumors （P〈0.05）. Conclusion： Tumor metastasis results from gradual accumulation of abnormal genetic alterations. Down-regulation of KISS-1 and up-regulation of S100A4 play a critical role in metastasis of gastric carcinoma.     

Expression of MMP-13 and its correlation with prognosis in non-small cell-lung cancer

Objective： To investigate the expression of MMP-13 in non-small cell lung cancer （NSCLC）, so as to analyze its correlation with prognosis of NSCLC. Methods： MMP-13 expression was detected in 99 NSCLC tissues and 32 normal lung tissues by immunohistochemical method. Results： （1） Expression of MMP-13 （51.5%, 51/99） in cancer tissues was significantly higher than that in normal tissues 0% （P 〈 0.05）. Expression level of MMP-13 was significantly related to lymph node metastasis and clinical stage （P 〈 0.01）. （2） Kaplan-Meier analysis showed that expression level of MMP-13 was closely cor- relate with the prognosis of NSCLC. Multivariate Cox model analysis suggested that the survival time was significantly related to clinical stage and the expression of MMP-13. Conclusion： MMP-13 is an independent factor that affect prognosis.     

P53、C-erbB-2和VEGF在非小细胞肺癌中的表达及其临床意义

Objective: To investigate the expressions and the clinical significance of P53, C-erbB-2 and vascular endothelial growth factor(VEGF)in non-small cell lung cancer(NSCLC).Methods: 121 specimens of NSCLC were examined for P53, C-erbB-2 and VEGF by immunohistochemical staining.Results: The positive rates of P53, C-erbB-2 and VEGF in the carcinomatous tissue were 43%, 39% and 31% respectively.P53 gene protein expression in lung cancer was significantly related to histological type and P-TNM staging of lung cancer patients(P＜0.05), and was not associated with the sex, age, the size of primary cancer, lymph node metastasis and cell differentiation(P＞0.05).C-erbB-2 gene protein expression in lung cancer was closely related to histological type and cell differentiation(P＜0.05), and was not associated with the sex, age, the size of primary cancer, lymph node metastasis and P-TNM staging of lung cancer patients(P＞0.05).VEGF in lung cancer was only closely related to cell differentiation(P＜0.05), and was not associated with the sex, age, the size of primary cancer, lymph node metastasis, histological type and P-TNM staging of lung cancer patients(P＞0.05).Conclusion: It is possible for P53, C-erbB-2 and VEGF to play an important role in the oncogenesis and development of non-small cell lung cancer.     

肝细胞癌组织中MMP-2的表达与侵袭转移的关系

OBJECTIVE：To investigate the association between MMP-2 expression of hepatocellular carcinoma （HCC） and its invasion and metastasis. METHODS： Semi-quantitative RT-PCR and immunohistochemical analysis technology were used to detect MMP-2 mRNA expression and MMP-2 protein expression respectively in HCC, non-HCC liver tissue samples and normal liver tissue samples. RESULTS： The level of MMP-2 expression in HCC was higher than that in adjacent non-HCC liver tissue and in normal liver tissue and the difference was statistically significant, P〈0.05. and the difference of MMP-2 expression between adjacent non-HCC liver tissue and normal liver tissue had no statistical significance, P〉0. 05. MMP-2 expression in HCC had an inverse correlation with recurrence after surgical treatment, P=0.013, R^2=0.842, and a posi- tive correlation with pathological grade, P = 0. 000, R^2 = 0. 797. There was a positive correlation between MMP-2 expression in HCC and extrahepatie metastasis, P=0. 003, R^ =0. 834. Significant inverse correlation was found between MMP-2 expression in adjacent non-HCC liver tissue and extrahepatie metastasis, P=0. 021, R^2= 0. 877. CONCLUSIONS： MMP-2 has an association with degree of tumor differentiation, capability of invasion and metastasis and tendency of recurrence. MMP-2 way be one of indexes of differentiation, recurrence and metastasis.