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Objective@#To investigate the expressions of hypoxia inducible factor-1α (HIF-1α) and autophagy related protein Beclin-1 in colon cancer and their clinical significances.@*Methods@#From January 2017 to December 2018, 120 patients with colon cancer who were admitted to Dandong First Hospital were selected. The expressions of HIF-1α and Beclin-1 in colon cancer and corresponding adjacent tissues were detected by immunohistochemical SABC method. The relationship of the expressions of HIF-1α and Beclin-1 with the clinicopathological features of colon cancer patients was also discussed. The expressions of HIF-1α and Beclin-1 in 20 pairs of fresh colon cancer and paracancerous tissues were detected by Western blot.@*Results@#The results of immunohistochemistry showed that the positive expression rates of HIF-1α and Beclin-1 proteins in colon cancer tissues were significantly higher than those in paracancerous tissues [75.0% (90/120) vs. 21.7% (26/120), 60.8% (73/120) vs. 12.5% (15/120)], and the differences were statistically significant (χ 2 values were 68.343 and 60.359, both P < 0.01). The results of Western blot showed that the expressions of HIF-1α and Beclin-1 proteins in 20 pairs of fresh colon cancer tissues were significantly higher than those in paracancerous tissues (1.98±0.66 vs. 0.76±0.55, 1.50±0.40 vs. 0.46±0.35), and the differences were statistically significant (t values were 6.912 and 8.315, both P < 0.01). The expressions of HIF-1α and Beclin-1 in colon cancer tissues were related to TNM stage, degree of differentiation, depth of infiltration, lymph node metastasis and nerve invasion (HIF-1α: χ 2 values were 9.074, 15.215, 10.101, 11.610 and 9.979; Beclin-1: χ 2 values were 11.285, 4.858, 24.436, 9.354 and 4.632; all P < 0.05), but not with age, sex, tumor location, tumor diameter and vascular tumor thrombus (all P > 0.05). There was a positive correlation between the expressions of HIF-1α and Beclin-1 proteins in colon cancer (r = 0.483, P < 0.01).@*Conclusion@#The expressions of HIF-1α and Beclin-1 proteins in colon cancer tissues are significantly increased, and the interaction between them is involved in the occurrence and development of colon cancer.
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Objective To investigate the expressions of Wnt3a and Wnt5a in papillary thyroid carcinoma (PTC) and their clinical significances. Methods Immunohistochemical SABC method was used to detect the expressions of Wnt3a and Wnt5a proteins in PTC tissues and their paracancerous tissues collected from 79 patients in Dandong First Hospital from January 2014 to June 2018, and the relationships between the expressions of Wnt3a and Wnt5a proteins and clinicopathological features of PTC patients were analyzed. The expressions of Wnt3a and Wnt5a proteins in 10 pairs of fresh PTC tissues and paracancerous tissues were detected by Western blot. Results The results of immunohistochemistry showed that the positive expression rates of Wnt3a and Wnt5a proteins in PTC tissues were significantly higher than those in paracancerous tissues [69.6％ (55/79) vs. 25.3％ (20/79), 60.8％ (48/79) vs. 20.2％ (16/79)], and the differences were statistically significant (χ 2 values were 31.092 and 26.894, both P < 0.01). The results of Western blot showed that the expressions of Wnt3a and Wnt5a proteins in 10 pairs of fresh PTC tissues was significantly higher than those in paracancerous tissues(1.61±0.40 vs. 0.43±0.14, 1.38±0.291 vs. 0.36±0.13), and the differences were statistically significant (t values were 16.234 and 13.493, both P < 0.01). The expressions of Wnt3a and Wnt5a in PTC tissues were correlated with TNM stage, differentiation, extramembranous invasion and lymph node metastasis (Wnt3a: χ2 values were 6.645, 15.945, 8.783 and 11.220; Wnt5a: χ2 values were 21.525, 7.611, 17.880 and 12.581, all P < 0.05), but not with patients'age, sex and tumor diameter (all P > 0.05). There was a positive correlation between Wnt3a and Wnt5a proteins expressions in PTC (r = 0.597, P < 0.01).Conclusion The abnormal expressions of Wnt3a and Wnt5a proteins in PTC may be related to the development of PTC.
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The changes in the expression of aquaporin-1 (AQP1) mRNA and protein in cultured human trabecular meshwork (HTM) cells treated with dexamethasone and transfected with antisense oligonucleotides (AS-ODN) were studied, and the implication of AQP1 regulation in corticosteroid-glaucoma and the possibility of AS-ODN inhibiting the AQP1 expression were evaluated.The cultured HTM cells in vitro were treated with different concentrations of dexamethasone and transfected with oligonucleotides for 5 days respectively. Then, total RNA and protein of HTM cells were extracted. The changes of AQP1 mRNA and protein were demonstrated qualitatively and quantitatively by RT-PCR and Western blot. Band intensities were detected by imaging analysis.There was a parallel relationship between the results of RT-PCR and those of Western blot. The expression levels of AQP1 mRNA and protein in dexamethasone-treated groups were increased initially and decreased later as dexamethasone concentration was stepped up. In the 0.04 μg/mL and 0.4 μg/mL groups, the levels of AQP1 were higher than in control group (0μg/mL). In the 4μg/mL and 40μg/mL groups, the AQP1 expression levels were lower than in control group. AS-ODN could down-regulate the expression of AQP1 mRNA and protein in a dose-dependent manner. At 5 μg/mL, down-regulation efficiency reached the maximum. There was no statistically significant difference in the expression of AQP1 mRNA and protein between all sense oligonucleotides groups and control group. It was suggested that dexamethasone may induce the changes of the AQP1 expression in HTM cells to be involved in the occurrence of corticosteroid-glaucoma. AS-ODN can down-regulate the AQP1 expression in HTM cells to some extent.
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In order to investigate the effect of nerve growth factor (NGF) on the proliferation of rabbit corneal endothelial cells and epithelial cells, the in vitro cultured rabbit corneal endothelial cells and epithelial cells were treated with different concentrations of NGF. MTT assay was used to examine the clonal growth and proliferation of the cells by determining the absorbency values at 570 nm. The results showed that NGF with three concentrations ranging from 5 U/mL to 500 U/mL enhanced the proliferation of rabbit corneal endothelial cells in a concentration-dependent manner. 50 U/mL and 500 U/mL NGF got more increase of proliferation than that of 5 U/mL NGF did. Meanwhile, 50 U/mL and 500 U/mL NGF could promote the proliferation of the rabbit corneal epithelial cells significantly in a concentration-dependent manner. However, 5 U/mL NGF did not enhance the proliferation of epithelial cells. It was suggested that exogenous NGF can stimulate the proliferation of both rabbit corneal endothelial and epithelial cells, but the extent of modulation is different.
Subject(s)
Cell Proliferation/drug effects , Cells, Cultured , Dose-Response Relationship, Drug , Endothelium, Corneal/cytology , Epithelium, Corneal/cytology , Nerve Growth Factor/pharmacologyABSTRACT
In order to investigate the effect of nerve growth factor (NGF) on the proliferation of rabbit corneal endothelial cells and epithelial cells, the in vitro cultured rabbit corneal endothelial cells and epithelial cells were treated with different concentrations of NGF.MTT assay was used to examine the clonal growth and proliferation of the cells by determining the absorbency values at 570nm. The results showed that NGF with three concentrations ranging from 5 U/mL to 500 U/mL enhanced the proliferation of rabbit corneal endothelial cells in a concentration-dependent manner.50 U/mL and 500 U/mL NGF got more increase of proliferation than that of 5 U/mL NGF did.Meanwhile, 50 U/mL and 500 U/mL NGF could promote the proliferation of the rabbit corneal epithelial cells significantly in a concentration-dependent manner. However, 5 U/mL NGF did not enhance the proliferation of epithelial cells. It was suggested that exogenous NGF can stimulate the proliferation of both rabbit corneal endothelial and epithelial cells, but the extent of modulation is different.
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The effects of suppression of CD44 by CD44-specific antisense oligonucleotide on attachment of human trabecular meshwork cells to hyaluronic acid (HA) were observed and the possible relationship between CD44 and primary open-angle glaucoma (POAG) investigated. CD44-specific antisense oligonucleotide was delivered with cationic lipid to cultured human trabecular meshwork cells. The expression of CD44 suppressed by CD44-specific antisense oligonucleotide was detected by RT-PCR and Western blotting. The effect of CD44 suppression by specific antisense oligonucleotide on attachment of trabecular meshwork cells to HA was measured by MTT assay. Results showed that expression of CD44 was suppressed by CD44-specific antisense oligonucleotide. Antisense oligonucleotide also suppressed the adhesion of human trabecular meshwork cells to HA in a concentration dependent manner. It was concluded that attachment of human trabecular meshwork cells to HA was decreased when CD44 was suppressed by specific antisense oligonucleotide. CD44 might play a role in pathogenesis of POAG by affecting the adhesion of trabecular meshwork cells to HA.
Subject(s)
Humans , Cell Adhesion , Cells, Cultured , Glaucoma, Open-Angle , Metabolism , Pathology , Hyaluronan Receptors , Genetics , Hyaluronic Acid , Metabolism , Oligonucleotides, Antisense , Pharmacology , Trabecular Meshwork , Cell BiologyABSTRACT
Objective To investigate the effect of turning scleral flap of self-body over drainage in trabeculectomy.Methods The surgery was performed on one eye of a rabbit while the other eye of the same rabbit was conducted with the routine trabeculectomy as the control group. The filtering bleb, intraocular pressure and pathology were observed. Results The intraocular pressure in the experimental eye was much lower than that in the control group 3～12 weeks after the operations (P＜0.05).As it was within 2 weeks after the surgeries, but there was no significant filtering difference (P＞0.05).The rate of eyes with functional bleb in the experimental group was significantly higher than that in the control group at the postoperative 2～7 weeks .Conclusion This study suggests the turning of self-body over drainage surgery can help to maintain filtering function for a long time and to prevent complication after filtering surgery.