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P2Y purinergic receptor activated PI-3K/Akt signaling pathway in regulation of growth and invasion of prostatic cancer.

Yu-xiang WANG ; Yong-hong SHI ; Li-hua GONG ; Yan LI ; Wan-jie HENG ; Jiang-feng YOU ; Hao-hao ZHONG ; Wei-gang FANG

Chinese Journal of Pathology.2007;36(10):681-686.

OBJECTIVETo investigate P2Y purinergic receptor activated PI-3K/Akt signaling pathway in the regulation of growth and invasion of prostate cancer in vitro.

METHODSWestern blot was used to detect phosphorylation of Akt (a downstream target molecule of PI-3K) by P2Y receptor agonist in 1E8 cells (a highly metastatic subclone derived from PC-3 prostatic cancer cell line). Cell counts, flow cytometry, Matrigel invasion assay, wound healing assay and gelatin zymography were used to detect changes of biological behaviors of 1E8 cells after P2Y receptor activation.

RESULTSAMP-PNP, one non-hydrolysis ATP analogue and P2Y receptor agonist, induced significant phosphorylation of Akt in a time- and dose-dependent manner in IE8 cells. LY294002, a specific inhibitor of PI-3K, effectively blocked Akt phosphorylation induced by AMP-PNP. Continuous exposure to AMP-PNP induced significant growth inhibition of 1E8 cells (inhibition rate at 50.2% at the 8th day), and this inhibition was mainly due to an arrest at S phase of the cell cycle (the S phase fraction of AMP-PNP treated cells was 22.3% higher than that of the control). Application of LY294002 did not reverse the growth inhibition effect of AMP-PNP. Matrigel invasion assay showed that AMP-PNP stimulation increased invasive ability of 1E8 cells, and this effect was effectively blocked by LY294002. No significant changes in the activation of MMP-2 and MMP-9 were detected by gelatin zymography, although wound healing assay showed 21.2% increase in cell migration after AMP-PNP treatment.

CONCLUSIONSPI-3K/Akt signaling pathway participates in P2Y receptor-stimulated prostate cancer invasion by enhancing cell motility, rather than up-regulating MMP-2 and MMP-9 activities. PI-3K signaling pathway plays an important role in prostate cancer proliferation, but is not involved in P2Y receptor mediated growth inhibition.

Adenylyl Imidodiphosphate ; pharmacology ; Animals ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Chromones ; pharmacology ; Humans ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Mice ; Mice, Nude ; Morpholines ; pharmacology ; Neoplasm Invasiveness ; Phosphatidylinositol 3-Kinases ; antagonists & inhibitors ; metabolism ; Phosphorylation ; Prostatic Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-akt ; metabolism ; Purinergic P2 Receptor Agonists ; S Phase ; drug effects ; Signal Transduction ; drug effects

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Expression of focal adhesion kinase in cardiac myocytes of hypertrophic ventricle.

Zhan-yu LI ; Xian-ping YI ; Ling ZHONG ; Fa-qian LI ; Wen-ying ZHOU ; Wan-wei CAO ; Yan-xuan ZHENG ; Xiao-hong WANG ; Ya-nan WANG

Chinese Journal of Pathology.2007;36(10):677-680.

OBJECTIVESTo investigate the role of focal adhesion kinase (FAK) in the pathogenesis of cardiac hypertrophy induced by hypertension.

METHODSUsing immunofluorescent labeling, confocal microscopy and Western blotting, the expression and subcellular localization of FAK in the cardiac myocytes of left ventricle were determined in 2, 6, 12, and 18 month-old rats with spontaneously hypertensive heart failure (SHHF) along with age-matched control Wistar-Kyoto (WKY) rats.

RESULTSThere was no significant difference of FAK expression between 2 month-old SHHF and WKY rats (50.5+/-6.9 vs. 49.8+/-5.0, n=6, P>0.05). In contrast with the control groups, the expression of FAK significantly increased in 6, 12 and 18 month-old SHHF rats (130.6+/-3.0 vs. 47.3+/-1.3, 144.7+/-5.4 vs. 46.4+/-3.1, 141.4+/-9.8 vs. 48.5+/-2.2, each groups n=6, P<0.05) with FAK protein primarily cumulated in the intercalated disks and nuclei.

CONCLUSIONSFAK may play a role in the cell signaling transduction leading to cardiac hypertrophy, presumably through regulations of hypertrophic gene transcription and RNA processing.

Animals ; Focal Adhesion Kinase 1 ; metabolism ; Heart Ventricles ; pathology ; Hypertension ; complications ; Hypertrophy, Left Ventricular ; enzymology ; etiology ; Male ; Microscopy, Confocal ; Myocytes, Cardiac ; metabolism ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Signal Transduction

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Overexpression of tumor metastasis suppressor gene 1 suppresses proliferation and invasion, but enhances apoptosis of human breast cancer cells MDA-MB-231 cells.

Jing SU ; Jiang-feng YOU ; Jie-liang WANG ; Xiang-lin CUI ; Wei-gang FANG ; Jie ZHENG

Chinese Journal of Pathology.2007;36(10):672-676.

OBJECTIVETo investigate the effects of tumor metastasis suppressor gene 1 (TMSG-1) overexpression on the proliferation, invasion and apoptosis of breast cancer cells and to determine possible correlations of TMSG-1 and metastasis of breast cancer.

METHODSFull-length human TMSG-1 coding sequences were cloned into plasmid pcDNA3.0-FLAG. The recombinant plasmids constructs were transfeced into MDA-MB-231, a highly malignant breast cancer cell line. Parental, vector-only stable transfectant and TMSG-1 stable transfectant clones were tested by MTT, soft agar colony formation and Boyden chamber assays. At twenty-four hours and forty-eight hours post transient transfection, double staining with Annexin-V-FITC and PI were employed to distinguish apoptotic cells from living cells by flow cytometry analysis.

RESULTSThree TMSG-1 overexpression clones were selected. Compared with the control cells, TMSG-1 overexpression MDA-MB-231 cells showed strong inhibition of proliferation and decreased clonogenicity in soft agar (P<0.05). Transfection of TMSG-1 into MDA-MB-231 cells significantly suppressed the cell invasion ability in vitro (decreased numbers of cells trespassing the matrigel in three experiments: 72.3+/-8.1, 85.0+/-4.2, and 73.5+/-7.8) in comparison with nave cells without transfection (187.5+/-2.1) and cells transfected with the control vector (162.3+/-6.8) (P<0.01). Transient transfection of TMSG-1 into MDA-MB-231 cells could promote cell apoptosis at 24 and 48 hours after transfection (P<0.05).

CONCLUSIONSTMSG-1 protein may have multiple functions in the regulation of proliferation, invasion and apoptosis of metastatic breast cancer cells, likely as a metastasis suppressor gene.

Apoptosis ; Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Membrane Proteins ; genetics ; metabolism ; physiology ; Neoplasm Invasiveness ; Plasmids ; Recombinant Proteins ; metabolism ; Sphingosine N-Acyltransferase ; genetics ; metabolism ; physiology ; Transfection ; Tumor Suppressor Proteins ; genetics ; metabolism ; physiology

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Clinicopathologic and prognostic study of pediatric immature teratoma.

Wen-ping YANG ; Yin ZOU ; Chuan-sheng HUANG ; Shu-zheng ZHANG ; Qiang XIAO ; Kang-lin DAI ; Hua-sheng ZHONG ; Xiao-jun XIONG

Chinese Journal of Pathology.2007;36(10):666-671.

OBJECTIVETo study the clinicopathologic features and biologic behavior of pediatric immature teratoma.

METHODSThe clinical data, pathologic features, immunohistochemical findings (for cyclin D1, P27 and Ki-67) and follow-up information of 39 cases of pediatric immature teratoma were analyzed.

RESULTSAmongst the 39 cases studied, 12 arose in the sacrococcygeal region, 12 in testis, 5 in retroperitoneum, 4 in ovary, 4 in abdomen and 2 in mediastinum. Histologically, 16 cases were of grade 1, 8 cases of grade 2 and 15 cases of grade 3. Seven of the cases contained foci of yolk sac tumor. Immature neuroepithelial features used in histologic grading included the presence of primitive neural tubules, immature rosettes, undifferentiated neuroblastoma cells and primitive neuroectodermal structures. Immunohistochemical study showed that cyclin D1 was positive in 3 cases of grade 1 tumors, 4 cases of grade 2 tumors and 9 cases of grade 3 tumors. The positivity rates for p27 were 8, 3 and 6 cases respectively, while those for Ki-67 were 3, 4 and 13 cases respectively. Follow-up data were available in 30 cases. Three of them, including 2 cases with histologic grade 3 (with or without yolk sac tumor component), recurred after operation.

CONCLUSIONSThe expression of cyclin D1 and Ki-67 is a useful adjunct in histologic grading. On the other hand, p27 overexpression shows little correlation with tumor grade. The prognosis of immature teratoma in children is different from that in adults. Sacrococcygeal immature teratoma occurring in patients younger than 1 year old and with low histologic grade do not require postoperative chemotherapy if the tumor is completely excised. Similarly, for testicular immature teratoma occurring in patients below 1 year of age, regardless of tumor grading, need no adjunctive therapy. On the other hand, ovarian immature teratoma with high histologic grade requires postoperative chemotherapy, regardless of age of the patients. The presence of microscopic foci of yolk sac tumor is a useful predictor of recurrence in pediatric immature teratoma.

Adolescent ; Cyclin D1 ; metabolism ; Endodermal Sinus Tumor ; drug therapy ; metabolism ; pathology ; surgery ; Female ; Follow-Up Studies ; Humans ; Infant ; Infant, Newborn ; Ki-67 Antigen ; metabolism ; Male ; Mediastinal Neoplasms ; drug therapy ; metabolism ; pathology ; surgery ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Ovarian Neoplasms ; drug therapy ; metabolism ; pathology ; surgery ; Proliferating Cell Nuclear Antigen ; metabolism ; Retroperitoneal Neoplasms ; drug therapy ; metabolism ; pathology ; surgery ; Sacrococcygeal Region ; Survival Rate ; Teratoma ; drug therapy ; metabolism ; pathology ; surgery ; Testicular Neoplasms ; drug therapy ; metabolism ; pathology ; surgery ; alpha-Fetoproteins ; metabolism

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Clinicopathologic study of intraabdominal extranodal follicular dendritic cell sarcoma.

Xiao-yu TU ; Wei-qi SHENG ; Hong-fen LU ; Jian WANG

Chinese Journal of Pathology.2007;36(10):660-665.

OBJECTIVETo study the clinicopathologic features and immunophenotype of intraabdominal extranodal follicular dendritic cell sarcoma (FDCS) and the relationship with Epstein-Barr virus (EBV).

METHODSThe clinical and histologic features of 4 cases of FDCS were evaluated. Immunohistochemical study was performed using standard EnVision method for CD21, CD23, CD35, S-100 protein, CD68, HLA-DR, vimentin, epithelial membrane antigen, desmin, CD34 and CD117. In-situ hybridization for EBV-encoded RNA (EBER) was carried out in 2 cases.

RESULTSThe age of patients ranged from 28 to 63 years (mean=42 years). The male-to-female ratio was 3:1. The clinical presentation was abdominal discomfort, pain or mass. Radiologic examination revealed concurrent lesions in stomach and left lobe of liver in 1 patient, while non-specific intraabdominal masses were detected in the remaining cases (in which the tumor was later found to be located in the appendix, mesentery of jejunum and omentum). Two cases were misdiagnosed as gastrointestinal stromal tumor before operation. Grossly, the tumors appeared as large solid nodules, with a mean diameter of 10.8 cm. Three of the cases showed areas of necrosis. Histologically, there were plump spindle, ovoid to epithelioid cells associated with scattered multinucleated giant cells. The tumor cells were arranged mostly in storiform pattern, whorls, fascicles or solid sheets. Lymphocytic infiltrates with perivascular cuffing were noted in all cases, resulting in a distinctive biphasic pattern. Two tumors showed significant cytologic atypia, with mitotic figures (including atypical mitotic figures) readily demonstrated. The remaining case (occurring in liver) was composed of scattered large atypical cells embedded in a dense inflammatory background, mimicking inflammatory pseudotumor. Immunohistochemical study showed that all cases were positive for CD21, CD23 and vimentin. There was focal expression of CD35, S-100 protein, CD68, HLA-DR and epithelial membrane antigen. The staining for CD34 and CD117 was negative. In-situ hybridization for EBER was negative in 2 cases tested.

CONCLUSIONSIntraabdominal extranodal FDCS is extremely rare. Familiarity with its characteristic histologic features and immunophenotype is important in distinguishing the tumor from other intraabdominal spindle cell lesions (such as gastrointestinal stromal tumor). Hepatic FDCS may show inflammatory pseudotumor-like features, resulting in misinterpretation. Non-hepatic intraabdominal FDCS seems to have little association with EBV infection.

Abdominal Neoplasms ; metabolism ; pathology ; virology ; Adult ; Dendritic Cell Sarcoma, Follicular ; metabolism ; pathology ; virology ; Diagnosis, Differential ; Epstein-Barr Virus Infections ; Female ; Gastrointestinal Stromal Tumors ; pathology ; Granuloma, Plasma Cell ; pathology ; Humans ; Immunophenotyping ; Male ; Middle Aged ; Receptors, Complement 3d ; metabolism ; Receptors, IgE ; metabolism ; Vimentin ; metabolism

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Subgrouping and outcome prediction of diffuse large B-cell lymphoma by immunohistochemistry.

Zi-yin YE ; Ya-bing CAO ; Tong-yu LIN ; Han-liang LIN

Chinese Journal of Pathology.2007;36(10):654-659.

OBJECTIVETo categorize diffuse large B-cell lymphoma (DLBCL) into germinal center B cell-like (GCB) and non-germinal center B cell-like (non-GCB) subgroups by immunohistochemistry; and to investigate the underlying prognostic significance.

METHODSImmunohistochemical study for CD10, bcl-6 and MUM1 was performed on 133 cases of DLBCL. The cases were then categorized into GCB and non-GCB subgroups. The 5-year overall survival and 5-year progression-free survival rates were compared between the GCB and non-GCB groups, and among the cases with different immunohistochemical expression or with different IPI.

RESULTSAmongst the 133 case studied, CD10 was expressed in 33.1%, while bcl-6 was positive in 34.6% and MUM1 in 45.1%. CD10 expression had a favorable impact on 5-year overall survival (P=0.041) and 5-year progression-free survival (P=0.031). On the other hand, bcl-6 expression had a favor able impact on 5-year progression-free survival (P=0.044). Expression of MUM1 carried an adverse effect on 5-year overall survival (P=0.031) and 5-year progression-free survival (P=0.028). GCB immunophenotype was demonstrated in 40.6% of the cases, while 59.4% showed a non-GCB profile. GCB DLBCL had a significantly longer 5-year overall survival (P=0.004) and 5-year progression-free survival (P=0.003), as compared with the non-GCB group. When dividing the cases into two groups according to their IPI score (IPI=0 to 1 and IPI=2 to 5), it turned out that the 5-year overall and progression-free survival rates of the GCB group were significantly higher than those of the non-GCB group (P=0.019 and 0.014 respectively in cases with IPI of 0 to 1 and P=0.006 and 0.009 respectively in cases with IPI of 2 to 5). The non-GCB cases with a IPI of 2 to 5 had the poorest prognosis.

CONCLUSIONDLBCL subgrouping by immunohistochemistry and analysis of the subgrouping with IPI is feasible and useful in predicting clinical outcome.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; B-Lymphocytes ; pathology ; Child ; Child, Preschool ; DNA-Binding Proteins ; metabolism ; Disease-Free Survival ; Female ; Follow-Up Studies ; Germinal Center ; pathology ; Humans ; Immunohistochemistry ; Interferon Regulatory Factors ; metabolism ; Kaplan-Meier Estimate ; Lymphoma, Large B-Cell, Diffuse ; classification ; immunology ; pathology ; Male ; Middle Aged ; Neprilysin ; metabolism ; Prognosis ; Proto-Oncogene Proteins c-bcl-6 ; Survival Rate ; Young Adult

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Application and evaluation of biologic potential grading criteria for localized gastrointestinal stromal tumors.

En-yi SHI ; Ying-yong HOU ; Yun-shan TAN ; Xiao-yan ZHOU ; Xu CAI ; Yong-ming LU ; Xiong-zeng ZHU

Chinese Journal of Pathology.2007;36(10):649-653.

OBJECTIVETo evaluate the implication of Fletcher and Miettinen biologic potential grading criteria in native localized gastrointestinal stromal tumors (GISTs).

METHODSTwo hundred and twenty localized GISTs with complete clinicopathologic and follow-up data were evaluated for their biologic potential by Fletcher and Miettinen grading criteria. The implication of the two grading criteria were compared by survival analysis.

RESULTSEvaluated by Fletcher grading criteria, the overall and disease-free survival rate of high risk GISTs was lower than that of very-low, low and intermediate GISTs; while the overall and disease-free survival rate of very-low, low and intermediate risk GISTs had no statistical diffence. In the high risk GISTs, the overall and disease-free survival rate of small intestinal and rectal GISTs was lower than that of gastric GISTs; while in the intermediate risk GISTs, the disease-free survival rate of small intestinal GISTs was lower than that of gastric GISTs. Evaluated by Miettinen grading criteria, the overall and disease-free survival rate of high risk GISTs was lower than that of very-low, low and intermediate GISTs; while the overall and disease-free survival rate of very-low, low and intermediate risk GISTs had no statistical difference. In the risk subgroup of GISTs, the overall and disease-free survival rate of gastric, small intestinal and rectal GISTs had no statistical difference.

CONCLUSIONSFletcher grading criteria is simple and easy to use; while Miettinen grading criteria for evaluating biological potential by anatomic site is more critical and has important reference implication for the selection of high risk patients for targeted adjuvant treatment.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Disease Progression ; Disease-Free Survival ; Female ; Follow-Up Studies ; Gastrointestinal Stromal Tumors ; pathology ; Humans ; Ileal Neoplasms ; pathology ; Jejunal Neoplasms ; pathology ; Kaplan-Meier Estimate ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Rectal Neoplasms ; pathology ; Risk Assessment ; methods ; standards ; Stomach Neoplasms ; pathology ; Survival Rate ; Young Adult

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Medulloblastoma with extensive nodularities: report of a case.

Qiu-ping GUI ; Xin SONG ; Huai-yu TONG

Chinese Journal of Pathology.2007;36(9):644-645.

Cerebellar Neoplasms ; diagnosis ; pathology ; radiotherapy ; surgery ; Follow-Up Studies ; Humans ; Infant ; Magnetic Resonance Imaging ; Male ; Medulloblastoma ; diagnosis ; pathology ; radiotherapy ; surgery

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Microglandular adenosis of breast: report of a case.

Gui-mei QU ; Zhi-qiang LANG ; Wei-dong YAO ; Guo-hua YU ; Wen-fang YU

Chinese Journal of Pathology.2007;36(9):643-644.

Anion Exchange Protein 1, Erythrocyte ; metabolism ; Breast ; metabolism ; pathology ; surgery ; Cadherins ; metabolism ; Female ; Fibrocystic Breast Disease ; metabolism ; pathology ; surgery ; Follow-Up Studies ; Humans ; Middle Aged ; S100 Proteins ; metabolism

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CD30-positive of diffuse large B-cell lymphoma of small intestine co-existing with tubular adenocarcinoma of rectum: report of a case.

Wei-hua LIU ; Xiao-ge ZHOU

Chinese Journal of Pathology.2007;36(9):641-642.

Adenocarcinoma ; metabolism ; pathology ; surgery ; Aged ; Humans ; Intestinal Neoplasms ; metabolism ; pathology ; surgery ; Intestine, Small ; Ki-1 Antigen ; metabolism ; Lymphoma, Large B-Cell, Diffuse ; metabolism ; pathology ; surgery ; Male ; Neoplasms, Multiple Primary ; metabolism ; pathology ; surgery ; Rectal Neoplasms ; metabolism ; pathology ; surgery

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