Runt-related transcription factor 3 (RUNX3) is a tumor suppressor gene whose reduced expression may play an important role in the development and progression of esophageal squamous cell cancer (ESCC). The aim of this study was to investigate the clinical relevance of RUNX3 in ESCC patients and effects of overexpression on biological behaviour of Eca109 cells in vitro and in vivo. Immunohistochemistry was performed to detect the clinical relevance of RUNX3 and lymph node metastasis in 80 ESCC tissues and 40 non-cancerous tissues using the SP method. RT-PCR and Western blotting were applied to assess the RUNX3 level and verify the Eca109 cell line with stable overexpression. Localization of RUNX3 proteins was performed by cell immunofluorescence. CCK-8 and Scrape motility assays were used to determine proliferation and migration and the TUNEL assay to analyze cell apoptosis. Invasive potential was assessed in cell transwell invasion experiments. In nude mice, tumorigenesis in vivo was determined. Results showed decreased expression of RUNX3 in esophageal tissue to be significantly related to lymph node metastasis (LNM) (P<0.01). In addition, construction of a recombinant lentiviral vector and transfection into the human ESCC cell line Eca109 demonstrated that overexpression could inhibit cell proliferation, migration and invasion, and induce apoptosis. The in vivo experiments in mice showed tumorigenicity and invasiveness to be significantly reduced. Taken together, our studies indicate that underexpression of RUNX3 in human ESCC tissue is significantly correlated with progression. Restoration of RUNX3 expression significantly inhibits ESCC cells proliferation, migration, invasion and tumorigenesis.
Overexpression of DNA methyltransferase 1 (DNMT1) has been detected in many cancers. Tobacco exposure is known to induce genetic and epigenetic changes in the pathogenesis of malignancy. 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) is an important carcinogen present in tobacco smoke; however the detailed molecular mechanism of how NNK induces esophageal carcinogenesis is still unclear. We found that DNMT1 was overexpressed in ESCC tissues compared with paired non-cancerous tissues, the overexpression being correlated with smoking status and low expression of $RAR{\beta}$. The latter could be upregulated by NNK treatment in Het-1A cells, and the increased DNMT1 expression level reflected promoter hypermethylation and downregulation of retinoic acid receptor ${\beta}$($RAR{\beta}$). RNA interference mediated knockdown of DNMT1 resulted in promoter demethylation and upregulation of $RAR{\beta}$ in KYSE30 and TE-1 cells. 3-(4,5-Dimethyl-thiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) and flow cytometric analysis demonstrated that NNK treatment in Het-1A cells could enhance cell proliferation and inhibit cell apoptosis in a dose-dependent manner. In conclusion, DNMT1 overexpression is correlated with smoking status and low expression of $RAR{\beta}$ in esophageal SCC patients. NNK could induce $RAR{\beta}$ promoter hypermethylation through upregulation of DNMT1 in esophageal squamous epithelial cells, finally leading to enhancement of cell proliferation and inhibition of apoptosis.
5-Azacytidine (5-azaC) was originally identified as an anticancer drug (NSC102876) which can cause hypomethylation of tumor suppressor genes. To assess its effects on runt-related transcription factor 3 (RUNX3), expression levels and the promoter methylation status of the RUNX3 gene were assessed. We also investigated alteration of biologic behavior of esophageal carcinoma TE-1 cells. MTT assays showed 5-azaC inhibited the proliferation of TE-1 cells in a time and dose-dependent way. Although other genes could be demethylated after 5-azaC intervention, we focused on RUNX3 gene in this study. The expression level of RUNX3 mRNA increased significantly in TE-1 cells after treatment with 5-azaC at hypotoxic levels. RT-PCR showed 5-azaC at $50{\mu}M$ had the highest RUNX3-induction activity. Methylation-specific PCR indicated that 5-azaC induced RUNX3 expression through demethylation. Migration and invasion of TE-1 cells were inhibited by 5-azaC, along with growth of Eca109 xenografts in nude mice. In conclusion, we demonstrate that the RUNX3 gene can be reactivated by the demethylation reagent 5-azaC, which inhibits the proliferation, migration and invasion of esophageal carcinoma TE-1 cells.
Zhou, Min;Yang, Li Jie;Yang, Wei Ren;Huang, Li Bo;Zhou, Xue Mei;Jiang, Shu Zhen;Yang, Zai Bin
Asian-Australasian Journal of Animal Sciences
/
v.31
no.1
/
pp.32-39
/
2018
Objective: In this study, we investigated the adverse effects of dietary zearalenone (ZEA) (0.5 to 1.5 mg/kg diet) on the localization and expression of the growth hormone receptor (GHR) in the uteri of post-weaning gilts and explored alternative mechanism of the reproductive toxicity of ZEA on piglets. Methods: A total of forty healthy piglets (Duroc${\times}$Landrace${\times}$Large White) aged 28 d were selected for study. Piglets were transferred to single cages after 10 days' adaptation on an obstetric table. The animals were allocated to one of four treatments: a normal basal diet supplemented with 0 (Control), 0.5 (ZEA0.5), 1.0 (ZEA1.0), or 1.5 (ZEA1.5) mg/kg purified ZEA, and fed for 35 d after the 10-d adaptation. Analyzed ZEA concentrations in the diets were 0, $0.52{\pm}0.07$, $1.04{\pm}0.03$, and $1.51{\pm}0.13mg/kg$, respectively. At the end of the feeding trial, piglets were euthanized after being fasted for 12 h. Two samples of uterine tissue from each pig were rapidly collected, one of which was stored at $-80^{\circ}C$ for analysis of the relative mRNA and protein expression of GHR, and the second was promptly fixed in Bouin's solution for immunohistochemical analysis. Results: The relative weight of the uteri and thickness of the myometrium and endometrium increased linearly (p<0.001) and quadratically (p<0.001) with an increasing level of ZEA. The results of immunohistochemical analysis indicated that GHR immunoreactive substance was mainly localizated in the cytoplasm of uterine smooth muscle, glandular epithelial, luminal epithelial, stromal, and vascular endothelial cells. In contrast, nuclear staining was rarely observed. The immunoreactive integrated optic density of GHR in the myometrium, luminal epithelium, glandular epithelium, and whole uteri of weaning gilts increased linearly (p<0.001) and quadratically (p<0.05) with an increasing level of ZEA. The mRNA and protein expression of GHR in the uteri of weaning gilts increased linearly (p<0.001) and quadratically (p<0.05) with an increasing level of ZEA. Conclusion: In conclusion, ZEA at a concentration of 0.5 mg/kg was sufficient to significantly thicken the myometrium and endometrium, and at a concentration of 1.0 mg/kg induced a high level of GHR expression to promote growth and development of the uteri. This revealed an alternative molecular mechanism whereby ZEA induces growth and development of the uteri and provides a theoretical basis for the revision of Chinese feed hygiene standards.
Objective: We previously found that the incidence of sarcopenia increased with declining glucose metabolism of muscle in patients with treatment-naïve diffuse large B-cell lymphoma (DLBCL). This study aimed to investigate the relationship between sarcopenia and muscle glucometabolism using 18F-FDG PET/CT at baseline and end-of-treatment, analyze the changes in these parameters through treatment, and assess their prognostic values. Materials and Methods: The records of 103 patients with DLBCL (median 54 years [range, 21-76]; male:female, 50:53) were retrospectively reviewed. Skeletal muscle area at the third lumbar vertebral (L3) level was measured, and skeletal muscle index (SMI) was calculated to determine sarcopenia, defined as SMI < 44.77 cm2/m2 and < 32.50 cm2/m2 for male and female, respectively. Glucometabolic parameters of the psoas major muscle, including maximum standardized uptake value (SUVmax) and mean standardized uptake value (SUVmean), were measured at L3 as well. Their changes across treatment were also calculated as ΔSMI, ΔSUVmax, and ΔSUVmean; Δbody mass index was also calculated. Associations between SMI and the metabolic parameters were analyzed, and their associations with progression-free survival (PFS) and overall survival (OS) were identified. Results: The incidence of sarcopenia was 29.1% and 36.9% before and after treatment, respectively. SMI (P = 0.004) was lower, and sarcopenia was more frequent (P = 0.011) at end-of-treatment than at baseline. The SUVmax and SUVmean of muscle were lower (P < 0.001) in sarcopenia than in non-sarcopenia at both baseline and end-of-treatment. ΔSMI was positively correlated with ΔSUVmax of muscle (P = 0.022). Multivariable Cox regression analysis showed that sarcopenia at end-of-treatment was independently negatively associated with PFS (adjusted hazard ratio [95% confidence interval], 2.469 [1.022-5.965]), while sarcopenia at baseline was independently negatively associated with OS (5.051 [1.453-17.562]). Conclusion: Sarcopenic patients had lower muscle glucometabolism, and the muscular and metabolic changes across treatment were positively correlated. Sarcopenia at baseline and end-of-treatment was negatively associated with the prognosis of DLBCL.
Seo, Jeong-Sik;Son, Su-Gyu;Kim, Ki-Sik;Seo, Sang-Myung;Kim, Dong-Han
Korean Journal of Medicinal Crop Science
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v.2
no.3
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pp.187-192
/
1994
This study was conducted to evaluate the effect of herbIcides on weed control, growth characteristics and yield in Angelica gigas NAKAI, after transplanting it to the field. All herbicids treated had no effect on the emergency period, bolting rate, and growth characteristics of A. gigos. Simazin wp, methabenzthiazuron wp were slightly harmful, but linuron wp pendimethalin ec, triflurain ec had no injury on the A. gigas, even with double dosage level. Under the treatment of pendimethalin ec, linulon wp, effect of weed control valuae at 55 days after transplanting was 89.6, and 88.3 respectively with 5 or 7 species of weeds appearing. Broadleaf weeds f?ere abundant than grasses, and the major weeds were C. albam, C. flexuos, D.sanguinalis, and E. arvense. Dry yield were increased somewhat more with herbicides than by hand weeding, even with double dosage level.
Kim, Doo-Youl;Kim, Kwang-Ho;Hong, Cheong-Ki;Lee, Seong-Yeul;Lee, Han-Bum;Huh, Beam-Lyang
KOREAN JOURNAL OF CROP SCIENCE
/
v.35
no.4
/
pp.342-351
/
1990
Experiments were conducted to find out the possibilities of producing good quality seed of summer type soybean at Chuncheon, 74m above sea level and Jinboo, 600m above sea level in 1987. Local variety, Cheonan Jaelae was used. Seeds were sown June 25 harvested five times from 35 days after flowering with five days interval to 55 days. Vegetative period was longer in the early planting and high cool area, Jinboo, which resulted longer stem length than the late planting and in the plain area, Chuncheon. Pod number and 100 grain weight were shown more and heavier in Jinboo. Earlier planting and later harvesting resulted more pod number and heaveier 100grain weight. Grain yield was heigher in Jinboo, and earlier plainting shown more grain yield and its difference was greater in Jinboo. Long grain filling duration resulted more grain yield. Unripened seeds were produced more by earlier planting with earlier harvesting and earlier planting with late harvesting showed the high rates of pod and stem blight (Diaporthe Phaseolorum). Seed With high moisture content by early harvesting showed more rotten seeds in all planting dates production was more advantageous in the high cool area, Jinboo. Optimum planting date was between May 15 and June 5. Harvesting was desirable about 45-50 days after flowering with heigher germination rate seed yield.
Man Lu;Liwei Xie;Sijie Yin;Jing Zhou;Lingmei Yi;Ling Ye
Journal of Microbiology and Biotechnology
/
v.34
no.9
/
pp.1769-1777
/
2024
Chemotherapy-induced nausea and vomiting (CINV) is a debilitating side effect related to activation of substance P (SP). SP activation can result from dysregulation of the gut-brain axis, and also from activation of protein kinase A signaling (PKA) signaling. In this study, we connected these factors in an attempt to unveil the mechanisms underlying CINV and develop new therapeutic strategies. Female rats were injected with cisplatin (Cis) to induce pica. Fecal samples were collected before/after injection, and subjected to lipid metabolomics analysis. In another portion of pica rats, the PKA inhibitor KT5720 was applied to investigate the involvement of PKA signaling in CINV, while fecal microbiota transplantation (FMT) was implemented to verify the therapeutic effect of the lipid metabolite 14(15)-EpETE. Pica symptoms were recorded, followed by ileal histological examination. The targeting relationship between 14(15)-EpETE and glucagon was determined by bioinformatics. SP and glucagon/PKA signaling in rat ileum, serum, and/or brain substantia nigra were detected by immunohistochemistry, enzyme-linked immunosorbent assay, and/or western blot. The results showed a significantly lower level of 14(15)-EpETE in rat feces after Cis injection. KT5720 treatment alleviated Cis-induced pica symptoms, ileal injury, SP content increase in the ileum, serum, and brain substantia nigra, and ileal PKA activation in rats. The ileal level of glucagon was elevated by Cis in rats. FMT exerted an effect similar to that of KT5720 treatment, relieving the Cis-induced changes, including ileal glucagon/PKA activation in rats. Our findings demonstrate that FMT restores 14(15)-EpETE production, which inhibits SP release by targeting GCG/PKA signaling, ultimately mitigating CINV.
This study was conducted to investigate the effects of Pleurotus eryngii fruiting body on blood glucose and cholesterol levels in streptozotocin-induced diabetic rats. Male Sprague-Dawley rats were used as experimental animals and experimental groups were divided into three groups, consisting of two control groups (diabetic and non-diabetic) and one P. eryngii group. Animals were fed experimental diets for 2 weeks. The values of died and water intake of P. eryngii group were lower than those of diabetic control group. The body weights of both groups after 2 weeks decreased by 18% as compared with initial values. The level of blood glucose decreased significantly by 16.9% in P. eryngii group as compared with diabetic control group, and there was a significant difference in glycosylated hemoglobin (GHb) level between both groups. The levels of total-cholesterol, LDL-cholesterol in plasma and atherosclerotic index were not significantly different between both groups. However the level of HDL-cholesterol increased significantly by 28% in P. eryngii group as compared with diabetic control group. These results suggested that fruiting bodies of P. eryngii exert blood glucose-lowering effect in streptozotocin-induced diabetic rats.
This study intends to analyze outcomes in labor market of students who graduated from provincial colleges. And For this aim, this study used 'Youth Panel Data' (2001, 2002) of HRD Korea Work Information Center. The reference groups are students who graduated from capital region colleges. The main results of this study are as follows. Students who graduated from provincial colleges have difficulty in acquiring job information than capital region college graduates. And students who graduated from provincial colleges get first job in smaller firm than capital region college graduates. The monthly wage level of students who graduated from provincial colleges is less than that of capital region college graduates. And the outcomes in labor market of students who graduated from Chungcheong region colleges are better than those of students who graduated from Yeongnam region and Honam region colleges. As results of this study show, there are differences between provincial college graduates and capital region college graduates. And there are differences between Chungcheong region college graduates and another region (namely, Yeongnam region and Honam region) college graduates.
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