PURPOSE: Angiogeneisis is essential process for tumor growth and metastasis. Vascular endothelial growth factor (VEGF) is major regulator of angiogenesis. Recently, the incidence of papillary thyroid microcarcinoma (PTMC) increased because of development in diagnostic modality. Several recent reports have documented relationship of VEGF and papillary thyroid cancer. The aims of this study were to determine whether angiogenetic phenotype was changed or not changed and to evaluate the relationship between clinicopathologic features and VEGF, vascular endothelial growth factor receptor-1 (VEGFR-1), hypoxic induced factor-1alpha (HIF-1α) mRNA expression in PTMC. METHODS: VEGF, VEGFR-1, HIF-1α mRNA expression was examined by RT-PCR in 14 patients who had undergone thyroidectomy due to PTMC. The thyroid tumor tissue and adjacent normal thyroid tissue were collected in operation and preserved at -70℃ in RNA later solution. We evaluate the expression of VEGF, VEGFR-1, HIF-1α mRNA by RTPCR. The expression of mRNA was quantititated by densitometer and analyzed the relationship between clinicopathologic features and mRNA expression. RESULTS: Compared to normal tissues, in PTMC we observed higher expression of HIF-1α mRNA (P=0.024) and lower expression of VEGF mRNA (P=0.002). There was no difference in expression of VEGFR-1.The patients with nodal metastasis had higher expression of the VEGF mRNA in tumor tissues than those without nodal metastasis but not significantly. The VEGF mRNA of tumor tissues in patients with thyroid capsule invasion or not were expressed similarly. The lower expression of VEGF mRNA were observed more frequently in younger patients (<40). CONCLUSION: The expression of VEGF mRNA was lower in tumor tissue in spite of higher expression of HIF-1α mRNA. These results suggest that the reason for good prognosis and no progression to clinical cancer in PTMC was related to the unchanged angiogenic phenotype.
Humans ; Incidence ; Neoplasm Metastasis ; Phenotype ; Prognosis ; RNA ; RNA, Messenger* ; Thyroid Gland* ; Thyroid Neoplasms ; Thyroidectomy ; Vascular Endothelial Growth Factor A* ; Vascular Endothelial Growth Factor Receptor-1*

PURPOSE: To gain insight on transcriptional repression of Topo II a in HL-60 cells arrested to G2/M and M phase, the levels of Topo IIa mRNA and the binding activity of ATF have been investigated with Northern blot hybridization and DNA mobility shift assay, respectively. MATERIALS AND METHODS: HL-60 cells were grown in RPMI 1640 medium supplemented with 10% heat-mactivated fetal bovine serum and antibiotics in a humidified 5% CO2 at 37C degree. Total RNA was prepared by a modification of the method of Karlinsey et al. Northern blot hybridization was performed by the method of Virca et al. A Xho I-Mlu I fragment of phTOP2 was used as probe for Northern blot analysis of Topo II a mRNA. DNA mobility shift assay was performed by the method of Lim et al. End labeled DNA oligomer (upper strand, 5-TCTCCGCTATGACGCCGAGTGGTG-3) for ATF binding activity was mixed with nuclear extracts in a 20 pl reaction volume containing 60 mM KC1, 12 mM HEPES, pH 7.9, 5 mM MgCl2, 0.2 mM EDTA, 0.2 mM DTT, 12% glycerol, and 2 ug of poly [dI-dC]. RESULTS: HL-60 cells were arrested at G2/M phase and M phase after taxol or nocodazole treatment. The levels of Topo II a mRNA were reduced at 24 hours after exposure with nocodazole or taxol but the unknotting activities were not changed. DNA mobility shift assay using oligonucleotide containing the ATF binding site showed that ATF binding activity was reduced after pretreatment of nododazole or taxol. CONCLUSIONS: These results suggest that the reduction of ATF binding activity may be important to transcriptional repression of Topo II a gene by nocodazole and taxol in HL- 60 cells.
Anti-Bacterial Agents ; Binding Sites ; Blotting, Northern ; Cell Division* ; DNA Topoisomerases, Type I* ; DNA Topoisomerases, Type II* ; DNA* ; Edetic Acid ; Electrophoretic Mobility Shift Assay ; Genes, vif ; Glycerol ; HEPES ; HL-60 Cells ; Humans ; Hydrogen-Ion Concentration ; Magnesium Chloride ; Nocodazole ; Paclitaxel ; Repression, Psychology ; RNA ; RNA, Messenger

Background: The Coronal Plane Alignment of the Knee (CPAK) classification system has been developed as a comprehensive system that describes 9 coronal plane phenotypes based on constitutional limb alignment and joint line obliquity (JLO). Due to the characteristics of Asian populations, which show more varus and wider distribution in lower limb alignment than other populations, modification of the boundaries of the arithmetic hip-knee-ankle angle (aHKA) and JLO should be considered. The purpose of this study was to determine the knee phenotype in a Korean population based on the original CPAK and modified CPAK classification systems. Methods: We reviewed prospectively collected data of 500 healthy and 500 osteoarthritic knees between 2021 and 2023 using radiographic analysis and divided them based on the modified CPAK classification system by widening the neutral boundaries of the aHKA to 0° ± 3° and using the actual JLO as a new variable. Using long-leg standing weight-bearing radiographs, 6 radiographic parameters were measured to evaluate the CPAK type: the mechanical HKA angle, medial proximal tibial angle (MPTA), lateral distal femoral angle (LDFA), aHKA, JLO, and actual JLO. Results: From 2 cohorts of 1,000 knees, the frequency distribution representing all CPAK types was different between the healthy and arthritic groups. The most common categories were type II (38.2%) in the healthy group and type I (53.8%) in the arthritic group based on the original CPAK classification. The left and upward shift in the distribution of knee phenotypes in the original classification was corrected evenly after re-establishing the boundaries of a neutral aHKA and the actual JLO. According to the modified CPAK classification system, the most common categories were type II (35.2%) in the healthy group and type I (38.0%) in the arthritic group. Conclusions Although the modified CPAK classification corrected the uneven distribution seen when applying the original classification system in a Korean population, the most common category was type I in Korean patients with osteoarthritic knees in both classification systems. Furthermore, there were different frequencies of knee phenotypes among healthy and arthritic knees.