OBJECTIVETo investigate the expression of cytokeratins and ret in thyroid papillary carcinoma (TPC) and their diagnostic value.

METHODSDuring the period of October 1999 to March 2002, 69 cases of TPC (42 cases with adjacent normal thyroid tissue) and 14 cases of nodular goiter with papillary hyperplasia were enrolled into the study. Immunohistochemistry for CK19, CK17, CK8, CK20 and ret was performed in all cases using EnVision and LSAB methods respectively.

RESULTSThe positive rates for CK19 and ret in TPCs were 85.5% and 68.1% respectively, which were significantly (P < 0.01) higher than those in nodular goiter and normal thyroid tissue (25.0% and 5.4% respectively). The expression of CK17 was also observed in a few cases of TPCs (11/69, 15.9%), which was mainly localized in areas of squamous metaplasia, poorly differentiated carcinoma and/or in the small infiltrative foci. The positive rates for CK8 were 75.4% and 26.8% in TPCs and benign thyroid tissue respectively. All cases were negative for CK20.

CONCLUSIONSCK19, CK17 and ret expressions are significantly higher in TPCs than benign thyroid tissue; and this characteristic can have important diagnostic value.

Carcinoma, Papillary ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Keratins ; analysis ; biosynthesis ; Proto-Oncogene Proteins ; biosynthesis ; Proto-Oncogene Proteins c-ret ; Receptor Protein-Tyrosine Kinases ; biosynthesis ; Thyroid Neoplasms ; metabolism ; pathology

The aim of study was to investigate the killing effect of double suicide gene system mediated by retroviral vector on K562 cells in vivo and ex vivo. CDglyTK gene was transfected into PA317 cells by using lipofectamine. K562 cells were infected with viral supernatant. K562/CDglyTK cells were treated with 5-fluorocytosine (5-FC) and/or ganciclovir (GCV). Mice were randomly divided into three groups: tumor formation, tumor inhibition and tumor therapy. Each mouse was implanted with K562/CDglyTK cells or K562 cells. The results indicated that the killing effect of 5-FC in combination with GCV on K562/CDglyTK was more significant than using 5-FC or GCV alone. In vivo study showed that after being injected subcutaneously with K562 cells and K562/CDglyTK cells, there was not obvious difference in tumor formation rate of mice, 5-FC + GCV could suppress tumor formation of the K562/CDglyTK cells. After being treated with 5-FC and GCV, the median tumor volume of mice implanted with K562/CDglyTK cells decreased obviously, compared with the control group. Their median survival was significantly prolonged. It is concluded that double suicide genes are more effective for killing effect on K562 cells in vivo and in ex vivo. It may be applicable to clinical gene therapy.
Cytosine Deaminase ; genetics ; Flucytosine ; pharmacology ; Ganciclovir ; pharmacology ; Genes, Transgenic, Suicide ; genetics ; Genetic Therapy ; Genetic Vectors ; genetics ; Humans ; K562 Cells ; Protein-Tyrosine Kinases ; genetics ; Receptor Protein-Tyrosine Kinases ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; genetics ; Recombination, Genetic ; Retroviridae ; genetics

The purpose of this study was to detect the serum PTK7 level of patients with acute lymphocytic leukemia, and to reveal its clinical value for diagnosis of diseases. A total of 136 patients diagnosed as acute lymphocytic leukemia from May 2012 to April 2014 in our hospital were enroled in this study and were divided into the L1 group (n = 42), L2 (n = 45) and L3 group (n = 49) according cytomorphology, and 48 normal children were selected as control group. Fluorescence quantitative PCR was used to detect mRNA level of PTK7 in peripheral blood mononuclear cells, and Western blot was used to detect PTK7 protein expression. The results showed that the PTK7 mRNA level in L1 group was significantly higher than that in normal group (P = 0.000) . The PTK7 mRNA level in L2 group was significantly higher than that in the L1 group (P = 0.000). The PTK7 mRNA level in L3 group and L2 group had not significantly different between each other (P = 0.123). Serum PTK7 protein level in L1 group was very significantly higher than that in normal group (P = 0.000) . The serum PTK7 protein level in L2 group were very significantly higher than that in the L1 group (P = 0.003) and serum PTK7 protein level in L3 and L2 group had no significance difference (P = 0.312) . It is concluded that the expression level of serum PTK7 protein has a potential clinical value for the diagnosis of acute lymphocytic leukemia, but without specificity for ALL subsets.
Cell Adhesion Molecules ; blood ; genetics ; Humans ; Leukocytes, Mononuclear ; metabolism ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; blood ; diagnosis ; genetics ; RNA, Messenger ; biosynthesis ; blood ; genetics ; Receptor Protein-Tyrosine Kinases ; blood ; genetics

Novel Oncogene with Kinase-domain (NOK) is a novel tumor-related gene, coding receptor like protein with a kinase domain. Overexpression of NOK leads to tumorigenesis and metastasis. To further study NOK function in physiological condition, it is necessary to prepare the anti-NOK antibody. In this report, GST fusion protein was adopted to prepare polyclonal antibodies against hNOK. The result showed that the antibodies we generated is with a very high titriation, and can be used for examination of NOK protein by Westernblot. Furthermore, the antibodies were used for immunohistochemistry in lung cancer tissues, and the results demonstrated high expression of hNOK in the tumor tissues. The antibody of hNOK we generated can serve as a diagnostic method for the lung cancer.
Animals ; Antibodies ; genetics ; metabolism ; Antibodies, Monoclonal ; biosynthesis ; genetics ; Biomarkers, Tumor ; genetics ; metabolism ; Humans ; Lung Neoplasms ; diagnosis ; genetics ; metabolism ; Mice ; Oncogene Proteins ; biosynthesis ; genetics ; immunology ; RNA, Messenger ; biosynthesis ; genetics ; Receptor Protein-Tyrosine Kinases ; biosynthesis ; genetics ; immunology

c-erbB-2 oncogene encodes a growth factor receptor whose amino acid sequence has extensive homology with human epidermal growth factor receptor. It is frequently overexpressed in human breast, ovary, lung, and stomach cancers, where its overexpression is related significantly to the prognosis. Tl investigate the possible role of c-erbB-2 oncogene in the oncogenesis of stomach cancer, we examined the genetic alterations of c-erbB-2 oncogene in 4 stomach cancer cell lines, SNU-1, SNU-5, SNU-16 and KATO III. There were no differences in c-erbB-2 mRNA level as well as c-erbB-2 gene copy number among them. But gp185-erbB-2, c-erbB-2 gene product, was increased from 2- to 4-fold in SNU-1 and SNU-5 cells, compared with that in SNU-16 or KATO III cells. Our results suggest that post-transcriptional regulation of gp185erbB-2 expression may underlie gp185erbB-2 overexpression in cancer cells.
Amino Acid Sequence ; Humans ; Molecular Sequence Data ; Protein-Tyrosine Kinases/*biosynthesis ; Proto-Oncogene Proteins/*biosynthesis/genetics/immunology ; RNA, Messenger/analysis ; Receptor, Epidermal Growth Factor/*biosynthesis/genetics/immunology ; Receptor, erbB-2 ; Receptors, Cell Surface/*biosynthesis ; Stomach Neoplasms/genetics/*metabolism ; Tumor Cells, Cultured

OBJECTIVETo analyze the clinicopathological characteristics of patients with anaplastic lymphoma kinase (ALK) rearrangements in lung adenocarcinoma, and the clinical therapy and prognosis of the patients.

METHODSClinicopathological data of 34 cases of ALK-positive patients treated in the Beijing Chest Hospital from 2005 to 2014 were reviewed. The expression of ALK proteins in the resected tumors was detected by immunohistochemistry, and EGFR mutations were examined by polymerase chain reaction and a direct DNA sequencing method.

RESULTSAmong the 34 patients, 20 were male and 14 were female, the median age was 49, and 11 were smokers and 23 were never smokers. The clinical stages of the patients were stage IA in 5 patients, IB in one patient, IIA in two patients, IIIA in 16 patients, IIIB in 5 patients, IV in 4 patients, and one patient of unknown stage. ALK-positive tumors showed strong granular staining in cell cytoplasm by immunohistochemistry. Forteen patients were solid predominant subtype with mucin production, 10 of acinar predominant subtype, 6 of papillary predominant subtype, 3 of micropapillary predominant subtype, and one was of colloid variant. There were 18 cases with mucin production, 6 cases had signet-ring cell morphology, and 10 cases showed cribriform pattern. Only one patient had coexistence of ALK rearrangement and EGFR mutation (L858R at exon 21). Of the 34 patients, 24 patients were followed up. The median follow up of the 24 patients was 11.0 months (1.7-48.7 months).

CONCLUSIONSALK-positive tumors as a molecular subtype of lung adenocarcinoma have distinct clinicopathological features. The histological findings of ALK-positive tumors are characterized by solid predominant subtype with mucin production, acinar predominant subtype, signet-ring cells and cribriform structures. They were rarely co-mutated with EGFR mutation.

Adenocarcinoma ; enzymology ; pathology ; therapy ; Exons ; Female ; Gene Rearrangement ; Genes, erbB-1 ; Humans ; Immunohistochemistry ; Lung Neoplasms ; enzymology ; pathology ; therapy ; Male ; Middle Aged ; Mucins ; biosynthesis ; Mutation ; Neoplasm Proteins ; genetics ; Polymerase Chain Reaction ; Prognosis ; Receptor Protein-Tyrosine Kinases ; genetics ; Sequence Analysis, DNA

Recently, the rearrangement of RET proto-oncogene has been reported to be the most common genetic change in papillary thyroid carcinoma (PTC). However, its prevalence has been reported variably and its relation to clinical outcome has been controversial. The characteristic nuclear features of PTC usually render the diagnosis, but problem arises with equivocal cytologic features that are present focally. Although there remains some controversy, CK19 has been reported to be a useful ancillary tool for diagnosis of PTC. To evaluate the expression rate of RET/PTC rearrangement and CK19 in PTCs in a Korean population, we studied 115 papillary thyroid carcinomas in 3 mm-core tissue microarray based immunohistochemical analysis. The prevalence of Ret protein expression was 62.6% and the CK19 immunoreactivity was 80.9%. There was no statistically significant asso-ciation between the Ret positivity and CK19 immunoreactivity, although the percent agreement of the two was relatively high. The clinicopathological variables did not correlate with the expression of Ret. In conclusion, the prevalence of Ret protein expression and its clinicopathological implications in a Korean population are not much different from those reported in previous studies. However, its detection via immunohistochemistry can be a useful diagnostic tool for diagnosing papillary thyroid carcinoma in conjunction with CK19.
Adenocarcinoma, Papillary/*metabolism ; Adult ; Carcinoma/pathology ; Cell Line, Tumor ; Cytoplasm/metabolism ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Immunohistochemistry ; Keratin/*biosynthesis ; Korea ; Lymphatic Metastasis ; Male ; Middle Aged ; Oligonucleotide Array Sequence Analysis ; Oncogene Proteins/*biosynthesis ; Receptor Protein-Tyrosine Kinases/*biosynthesis ; Thyroid Neoplasms/*metabolism/pathology

Lysophosphatidic acid (LPA) is a bioactive phospholipids and involves in various cellular events, including tumor cell migration. In the present study, we investigated LPA receptor and its transactivation to EGFR for cyclooxygenase-2 (COX-2) expression and cell migration in CAOV-3 ovarian cancer cells. LPA induced COX-2 expression in a dose-dependent manner, and pretreatment of the cells with pharmacological inhibitors of Gi (pertussis toxin), Src (PP2), EGF receptor (EGFR) (AG1478), ERK (PD98059) significantly inhibited LPA- induced COX-2 expression. Consistent to these results, transfection of the cells with selective Src siRNA attenuated COX-2 expression by LPA. LPA stimulated CAOV-3 cell migration that was abrogated by pharmacological inhibitors and antibody of EP2. Higher expression of LPA2 mRNA was observed in CAOV-3 cells, and transfection of the cells with a selective LPA2 siRNA significantly inhibited LPA-induced activation of EGFR and ERK, as well as COX-2 expression. Importantly, LPA2 siRNA also blocked LPA-induced ovarian cancer cell migration. Collectively, our results clearly show the significance of LPA2 and Gi/Src pathway for LPA-induced COX-2 expression and cell migration that could be a promising drug target for ovarian cancer cell metastasis.
Butadienes/pharmacology ; Cell Line, Tumor ; Cell Movement/drug effects/*physiology ; Cyclooxygenase 2/*biosynthesis ; Extracellular Signal-Regulated MAP Kinases/antagonists & inhibitors/metabolism ; Female ; Flavonoids/pharmacology ; GTP-Binding Protein alpha Subunits, Gi-Go/antagonists & inhibitors/*metabolism ; Humans ; Lysophospholipids/pharmacology ; Nitriles/pharmacology ; Ovarian Neoplasms/metabolism/*pathology ; Pertussis Toxin/pharmacology ; Protein-Tyrosine Kinases/antagonists & inhibitors/*metabolism ; Proto-Oncogene Proteins/antagonists & inhibitors/*metabolism ; Pyrimidines/pharmacology ; Receptor, Epidermal Growth Factor/antagonists & inhibitors/metabolism ; Receptors, Lysophosphatidic Acid/*metabolism ; Receptors, Prostaglandin E/metabolism ; Signal Transduction ; Transcriptional Activation ; Tyrphostins/pharmacology

Leptin, the product of ob gene, is an endocrine hormone that regulates adipose tissue mass. Recently, leptin has been found to generate a growth signal involving a tyrosine kinase-dependent intracellular pathway and promote angiogenic processes via activation of leptin receptor (Ob-R) in endothelial cells. However, it is not clear how leptin functions to promote multi-step processes involved in the neovascularization at the atherosclerotic plaque. We have examined the expression of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) and Ob-R in human atherosclerotic lesions, leptin-mediated angiogenesis in vivo and in vitro. Immunohistochemical analysis of human atherosclerotic aorta revealed an increased expression of Ob-R in the intima of neorevascularized regions and of both MMPs and TIMPs predominantly in the endothelial lining of intimal neovessels and macrophages/foam cells. In the rat corneal angiogenesis assay, leptin elicited a comparable sensitivity of angiogenic activity to those of vascular endothelial growth factor (VEGF). The immunohistological analysis of the leptin-treated rat cornea showed definitive rises in Ob-R, MMPs and TIMPs expression as well as those of VEGF receptor (VEGFR-1). Leptin (10-40 ng/ml) induced proliferation of the human umbilical vein endothelial cells (HUVECs) and elevation of MMP-2, MMP-9, TIMP-1, and TIMP-2 expression in a dose-dependent manner. Leptin also induced increases of MMP-2, MMP-9, TIMP-1, and Up-regulated the human coronary artery smooth muscle cells (HCASMCs). These findings suggest that leptin, a hormone with pluralistic properties including a mitogenic activity on vascular endothelial cells, plays a role in matrix remodeling by regulating the expression of MMPs and TIMPs. Taken together, our findings further provide evidences for leptin's role as an angiogenesis inducer in the normal organ (rat cornea) and in aberrant vasculature under duress like atherosclerosis.
Animal ; Arteriosclerosis/metabolism ; Blotting, Western ; Cell Division ; Cells, Cultured ; Dose-Response Relationship, Drug ; Endothelial Growth Factors/metabolism ; Endothelium, Vascular/*cytology/*enzymology ; Enzyme-Linked Immunosorbent Assay ; Fibroblast Growth Factor 2/metabolism ; Immunohistochemistry ; Leptin/*chemistry/metabolism/*physiology ; Lymphokines/metabolism ; Matrix Metalloproteinases/*biosynthesis ; *Neovascularization, Pathologic ; Rats ; Receptor Protein-Tyrosine Kinases/metabolism ; Receptors, Growth Factor/metabolism ; Recombinant Proteins/metabolism ; Support, Non-U.S. Gov't ; Tissue Inhibitor of Metalloproteinases/metabolism ; Umbilical Veins/metabolism ; Up-Regulation