Molecular and Immunohistochemical Expression of LTA4H and FXR1 in Canine Oral Melanoma

Oral melanoma is a common canine tumor whose prognosis is considered ominous, but poorly predicted by histology alone. In the present study the gene and protein expression of Leukotriene A4 hydrolase (LTA4H) and Fragile-X-mental retardation-related protein1 (FXR1), both reported as related to metastatic potential in different tumors, were investigated in canine oral melanoma. The main aim of the study was to confirm and quantify the presence of LTA4H and FXR1 genes and protein in oral melanomas. A secondary aim was to investigate their association with histologic prognostic criteria (mitotic count, Ki-67 index). Formalin-fixed-paraffin-embedded canine oral melanomas (36) were collected and histopathological evaluation carried out. Immunolabelling for LTA4H and FXR1 and Ki-67 were performed. RT-PCR evaluated LTA4H and FXR1 gene expressions. Histologically, most tumors were epithelioid cell melanomas (19/36) and were amelanotic, mildly or moderately pigmented (5, 12 and 13/36 respectively), only 6 were highly pigmented. Mitotic count ranged 1-106, Ki-67 index ranged 4.5–52.3. Thirty-two (32/32) melanomas immunolabelled for LTA4H and 33/34 for FXR1. RT-PCR values ranged 0.76–5.11 ΔCt for LTA4H and 0.22–6.24 ΔCt for FXR1. Molecular and immunohistochemical expression of both LTA4H and FXR1 did not statically correlate with mitotic count or Ki-67 index. The present study demonstrates LTA4H and FXR1 gene and protein in canine oral melanoma, however their expression is apparently unrelated to histopathologic prognostic criteria. Although LTA4H and FXR1 seem unrelated to tumor behavior, their extensive expression in the present cohort of cases suggest that they may play a role in canine oral melanoma oncogenesis.

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Neural differentiation of adipose-derived stem cells by indirect co-culture with Schwann cells

Archives of Biological Sciences ◽

10.2298/abs0904703l ◽

2009 ◽

Vol 61 (4) ◽

pp. 703-711 ◽

Cited By ~ 5

Author(s):

Xiaojie Li ◽

Dapeng Liao ◽

Ping Gong ◽

Quan Yuan ◽

Zhen Tan

Keyword(s):

Stem Cells ◽

Schwann Cell ◽

Neural Differentiation ◽

Adipose Derived Stem Cells ◽

Rt Pcr ◽

Gene Expressions ◽

Qrt Pcr ◽

Gene And Protein Expression ◽

Transwell Culture ◽

S 100

To investigate whether adipose-derived stem cells (ADSCs) could be subject to neural differentiation induced only by Schwann cell (SC) factors, we co-cultured ADSCs and SCs in transwell culture dishes. Immunoassaying, Western blot analysis, and RT-PCR were performed (1, 3, 7, 14 d) and the co-cultured ADSCs showed gene and protein expression of S-100, Nestin, and GFAP. Further, qRT-PCR disclosed relative quantitative differences in the above three gene expressions. We think ADSCs can undergo induced neural differentiation by being co-cultured with SCs, and such differentia?tions begin 1 day after co-culture, become apparent after 7 days, and thereafter remain stable till the 14th day.

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Inhibition of Migration and Invasion of Hepatocarcinoma HepG2 Cells by Dietary Saponins via Targeting HIF-1α

Anti-Cancer Agents in Medicinal Chemistry ◽

10.2174/1871520618666180110141827 ◽

2018 ◽

Vol 18 (7) ◽

pp. 1025-1031

Author(s):

Cheng Luo ◽

Di Wu ◽

Meiling Chen ◽

Wenhua Miao ◽

Changfeng Xue ◽

...

Keyword(s):

Cell Proliferation ◽

Confocal Microscopy ◽

Protein Expression ◽

Mtt Assay ◽

Hepg2 Cells ◽

Molecular Mechanisms ◽

Migration And Invasion ◽

Rt Pcr ◽

Gene And Protein Expression ◽

Simulated Hypoxia

Background: Different saponins from herbs have been used as tonic or functional foods, and for treatment of various diseases including cancers. Although clinical data has supported the function of these saponins, their underlying molecular mechanisms have not been well defined. Methods: With the simulated hypoxia created by 8 hours of Cu++ exposure and following 24 hour incubation with different concentration of saponins in HepG2 cells for MTT assay, migration and invasion assays, and for RT-PCR, and with each group of cells for immunofluorescence observation by confocal microscopy. Results： ZC-4 had the highest rate of inhibition of cell proliferation by MTT assay, and the highest inhibition of migration rate by in vitro scratch assay, while ZC-3 had the highest inhibition of invasion ratio by transwell assay. Under the same simulated hypoxia, the molecular mechanism of saponin function was conducted by measuring the gene expression of Hypoxia Inducible Factor (HIF)-1α through RT-PCR, in which ZC-3 showed a potent inhibition of gene HIF-1α. For the protein expression by immunofluorescence staining with confocal microscopy, HIF-1α was also inhibited by saponins, with the most potent one being ZC-4 after eight hours’ relatively hypoxia incubation. Conclusion: Saponins ZC-4 and ZC-3 have the potential to reduce HepG2 cell proliferation, migration and invasion caused by hypoxia through effectively inhibiting the gene and protein expression of HIF-1α directly and as antioxidant indirectly

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Dynamic changes of podocytes caused by fibroblast growth factor 2 in culture

Cell and Tissue Research ◽

10.1007/s00441-021-03511-x ◽

2021 ◽

Author(s):

Eishin Yaoita ◽

Masaaki Nameta ◽

Yutaka Yoshida ◽

Hidehiko Fujinaka

Keyword(s):

Growth Factor ◽

Fibroblast Growth Factor ◽

Fibroblast Growth Factor 2 ◽

Quiescent State ◽

Fibroblast Growth ◽

Gene Expressions ◽

Dynamic Changes ◽

Ki 67

AbstractFibroblast growth factor 2 (FGF2) augments podocyte injury, which induces glomerulosclerosis, although the mechanisms remain obscure. In this study, we investigated the effects of FGF2 on cultured podocytes with interdigitating cell processes in rats. After 48 h incubation with FGF2 dynamic changes in the shape of primary processes and cell bodies of podocytes resulted in the loss of interdigitation, which was clearly shown by time-lapse photography. FGF2 reduced the gene expressions of constituents of the slit diaphragm, inflections of intercellular junctions positive for nephrin, and the width of the intercellular space. Immunostaining for the proliferation marker Ki-67 was rarely seen and weakly stained in the control without FGF2, whereas intensely stained cells were frequently found in the presence of FGF2. Binucleation and cell division were also observed, although no significant increase in cell number was shown. An in vitro scratch assay revealed that FGF2 enhanced migration of podocytes. These findings show that FGF2 makes podocytes to transition from the quiescent state into the cell cycle and change their morphology due to enhanced motility, and that the culture system in this study is useful for analyzing the pathological changes of podocytes in vivo.

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Protective role of Sterculia tragacantha aqueous extract on pancreatic gene expression and oxidative stress parameters in streptozotocin-induced diabetic rats

Journal of Complementary and Integrative Medicine ◽

10.1515/jcim-2021-0020 ◽

2021 ◽

Vol 0 (0) ◽

Author(s):

Basiru Olaitan Ajiboye ◽

Babatunji Emmanuel Oyinloye ◽

Jennifer Chidera Awurum ◽

Sunday Amos Onikanni ◽

Adedotun Adefolalu ◽

...

Keyword(s):

Oxidative Stress ◽

Body Weight ◽

Protective Role ◽

Diabetic Rats ◽

Gene Expressions ◽

Ki 67 ◽

Oxidative Stress Parameters ◽

And Oxidative Stress ◽

Stress Parameters

Abstract Objectives The current study evaluates the protective role of aqueous extract of Sterculia tragacantha leaf (AESTL) on pancreatic gene expressions (insulin, PCNA, PDX-1, KI-67 and GLP-1R) and oxidative stress parameters in streptozotocin-induced diabetic rats. Methods Diabetes mellitus was induced into the experimental Wistar animals via intraperitoneal (IP) injection of streptozotocin (35 mg/kg body weight) and 5% glucose water was given to the rats for 24 h after induction. The animals were categorized into five groups of 10 rats each as follows normal control, diabetic control, diabetic rats administered AESTL (150 and 300 mg/kg body weight) and diabetic rats administered metformin (200 mg/kg) orally for two weeks. Thereafter, the animals were euthanized, blood sample collected, pancreas harvested and some pancreatic gene expressions (such as insulin, PCNA, PDX-1, KI-67, and GLP-1R)s as well as oxidative stress parameters were analyzed. Results The results revealed that AESTL significantly (p<0.05) reduced fasting blood glucose level, food and water intake, and lipid peroxidation in diabetic rats. Diabetic rats administered different doses of AESTL showed a substantial upsurge in body weight, antioxidant enzyme activities, and pancreatic gene expressions (insulin, PCNA, PDX-1, KI-67, and GLP-1R). Conclusions It can therefore be concluded that AESTL has the ability to protect the pancreas during diabetes mellitus conditions.

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Immunohistochemical expression of HER ‐ 2 and Ki ‐ 67 in granulosa cell tumor in bitches

Reproduction in Domestic Animals ◽

10.1111/rda.13903 ◽

2021 ◽

Author(s):

Ana Carina Heil da Silva Matos ◽

Angélica Consalter ◽

Bárbara Paula Santos Batista ◽

Ana Beatriz Monteiro Fonseca ◽

Ana Maria Reis Ferreira ◽

...

Keyword(s):

Granulosa Cell ◽

Granulosa Cell Tumor ◽

Cell Tumor ◽

Immunohistochemical Expression ◽

Ki 67 ◽

Her 2

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Analysis of the immunohistochemical expressions of p53, bcl-2 and Ki-67 in colorectal adenocarcinoma and their correlations with the prognostic factors

Arquivos de Gastroenterologia ◽

10.1590/s0004-28032010000200005 ◽

2010 ◽

Vol 47 (2) ◽

pp. 141-147 ◽

Cited By ~ 19

Author(s):

Hunaldo Lima de Menezes ◽

Mário Jorge Jucá ◽

Edmundo Guilherme de A. Gomes ◽

Benício L. Bulhões B. P. Nunes ◽

Henrique Oliveira Costa ◽

...

Keyword(s):

Prognostic Factors ◽

Cell Differentiation ◽

Colorectal Adenocarcinoma ◽

Sao Paulo ◽

Immunohistochemical Expression ◽

Ki 67 ◽

Disease Free Interval ◽

Free Interval ◽

Specific Mortality ◽

Disease Free

CONTEXT: Search of tumors markers that allow treatment with higher survival rates, and indicate the response to treatment and recurrence of cancer OBJECTIVE: To analyze the immunoexpression of the proteins p53, bcl-2 and Ki-67 in colorectal adenocarcinoma and correlate them with the clinical-pathological prognostic factors. METHOD: Tissue microarray paraffin blocks were made from colorectal adenocarcinoma tissue resected from 82 patients who had undergone surgery but not chemotherapy or radiotherapy, at "Hospital São Paulo", São Paulo, SP, Brazil, between 2002 and 2005. Thin sections (4 µm) were subjected to immunohistochemical reactions, and immunoexpression staining scores were obtained. The scores were correlated with the degree of cell differentiation, staging, disease-free interval, recurrence, survival and specific mortality. The study variables were analyzed using the chi-square and Kaplan-Meier tests to investigate associations with the markers. The significance of the differences between the curves of the disease-free interval and survival was analyzed using the Logrank and Wilcoxon tests. RESULTS: The immunohistochemical expression of p53 was positive in 70 tumors (85.4%) and negative in 12 (14.6%). The expression of bcl-2 was positive in 26 (31.7%) and negative in 56 (68.3%). The expression of Ki-67 was positive in 62 (75.6%) and negative in 20 (24.4%). There was no statistically significant correlation between the expressions of these markers separately or in conjunction, in relation to the degree of cell differentiation, staging, disease-free interval, survival and specific mortality. In relation to recurrence, there was a statistically significant correlation with positive expression of Ki-67 (P = 0.035). CONCLUSION: The immunohistochemical expression of Ki-67 in colorectal cancer is associated with recurrence of this disease.

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Expression of Grb7 growth factor receptor signaling protein in kidney development and in adult kidney

AJP Renal Physiology ◽

10.1152/ajprenal.1998.275.5.f770 ◽

1998 ◽

Vol 275 (5) ◽

pp. F770-F776 ◽

Cited By ~ 2

Author(s):

Sean F. Leavey ◽

Lois J. Arend ◽

Heidi Dare ◽

Gregory R. Dressler ◽

Josie P. Briggs ◽

...

Keyword(s):

Gene Expression ◽

Growth Factor ◽

Kidney Development ◽

Basic Function ◽

Growth Factor Signaling ◽

Receptor Signaling ◽

Rt Pcr ◽

Signaling Protein ◽

Gene And Protein Expression ◽

Grb7 Gene

Grb7, a signaling protein whose physiological function is unknown, binds receptor tyrosine kinases important for normal kidney development. By investigating and correlating Grb7 gene expression with that reported for Grb7-binding receptors, we provide clues to Grb7 function(s). RT-PCR and immunoblot were used to demonstrate Grb7 gene and protein expression in the mature kidney. Additional RT-PCR studies detected gene expression in all microdissected adult nephron segments examined, except glomeruli, and in the mouse metanephric kidney from embryonic day 11( E11) through to day 17( E17). In situ hybridization at E14 demonstrated the following cellular pattern of localization: Grb7 mRNA in metanephric epithelia of mesenchymal and ureteric bud origin; no expression in the undifferentiated mesenchyme; and little expression in podocyte-destined cells or primitive glomeruli. Grb7 mRNA was also present in the epithelia of the lung and gut at E14. Thus Grb7 may have a basic function in growth factor signaling in terminally differentiated epithelia along the nephron and in developing epithelia in the kidney, lung, and gut. It is localized in a pattern permissive for a role in Her2 and Ret receptor signaling.

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