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Objective:To explore the expression of long-chain noncoding RNA (lncRNA) and myocardial infarction-associated transcription (MIAT) in Leukocyte differentiation antigen (CD)4+T cells in peripheral blood of gastric cancer patients and its value of clinical application.Methods:Peripheral blood CD4+T cells were collected from 124 patients with gastric cancer, 90 benign gastric diseases patients and 80 healthy controls enrolled in Taizhou People′s Hospital from January 2019 to April 2021. The expression levels of MIAT and N6-methyladenosine(m6A) binding to MIAT promoter in CD4+T cells were detected by real-time fluorescent quantitative polymerase chain reaction (qPCR) and Chromatin immunoprecipitation (ChIP)-qPCR, respectively. Spearman test was used to analyze the correlation between MIAT and clinicopathological features, as well as between MIAT and regulatory T cell levels. The receivor operating characteristic curve (ROC) of the subjects was used to evaluate the MIAT expression level in the auxiliary diagnostic value of gastric cancer.Results:The relative expression levels of MIAT in the gastric cancer patients, the benign gastric diseases patients, and the healthy controls were 2.849 (2.131, 4.062), 1.511 (0.916, 1.855) and 0.963 (0.729, 1.432), respectively. The difference among the three groups was statistically significant ( H=158.25, P<0.001). The relative expression level of MIAT in the gastric cancer patients was significantly higher than the levels in the benign gastric diseases patients and healthy controls. The difference was statistically significant ( Z=100.63, 145.14, P<0.001). The binding activity of m6A to MIAT promoter in patients with early stage (stage Ⅰ and Ⅱ) and end stage (stage Ⅲ and Ⅳ) gastric cancer was 8.590±1.483 and 4.274±0.425, respectively. The difference was statistically significant ( t=6.255, P=0.002). Furthermore, the binding activity of m6A to MIAT promoter in the gastric cancer patients was significantly lower than that in patients with benign gastric diseases (17.267±3.106) and healthy controls (27.637±3.945) ( t=-7.331,-12.832, P<0.001). The relative expression of MIAT in CD4+T cells in peripheral blood of the gastric cancer patients had no significant difference in age(χ2=0.000, P=1.000), gender(χ2=0.000, P=1.000), CEA (χ2=0.648, P=0.421) and CA199(χ2=1.554, P=0.213), but had significant difference with tumor size expression(χ2=9.443, P<0.01), TNM stage(χ2=23.571, P=0.002) and lymph node metastasis (χ2=45.248, P<0.01). In addition, there was a significant positive correlation between the relative expression of MIAT in CD4+T cells and Treg level ( r2=0.76, P<0.001). The diagnostic efficacies of MIAT in CD4+T cells, CEA and CA199 in the gastric cancer patients were analyzed by ROC curve. When compared with patients with benign gastric diseases, the areas under the curve were 0.879, 0.635 and 0.611, respectively. When compared with healthy patients, the areas under the curve were 0.953, 0.784 and 0.598, respectively. Conclusions:The level of MIAT in CD4+T cells in peripheral blood of patients with gastric cancer is significantly higher than the levels in patients with benign gastric diseases and the healthy controls, which may be related with the decreased activity of m6A binding to the promoter of MIAT. The level of MIAT in CD4+T cells may be a relevant biomarker for the diagnosis and prognosis of gastric cancer.
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Objective Recent evidence has implicated Larp1,an RNA-binding protein,in cell motility and EMT therefore prompting the study of Larp1 in EOC.This project aims to examine the potential role of Larp1 in cell invasion.Methods Ovarian cancer cells SKOV-3 and OVCAR-3 were transfected with DNA constructs to overexpress Larp1 and transfect cells were used to assess the overexpression of Larp1 in cell invasion and metastasis using several functional assay.Results Larp1 overexpression in both OVCAR-3 and SKOV-3 cells appear to enhance the invasive ability of cells and the accumulation of Larp 1 in these chemoinvasive protrusions strongly suggests the potential role of Larp1 in invadopodia formation.Cell spot chemoinvasion assay demonstrated increased chemoinvasion of Larp1 overexpressing SKOV-3 and OVCAR-3 cells towards a gradient of TGF-β,EGF and bFGF.Conclusions With its potential role in EMT and cell invasion,a deeper understanding of the physiological role of Larp1 in cancer cell motility will be potentially beneficial in the diagnosis and treatments of ovarian cancer.
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Objective To investigate the role of the activated rhuPAa-melittin in ovarian cancer cells and to study the inhibitory effect of rhuPAa-melittinon on ovarian cancer cells. Methods rhuPAa-melittin was used to treat the ovarian cancer cells at different concentrations for 48 hrs. Then flow cytometery was applied to detect the cell cycle and cell apoptosis. rhuPAa-melittin protein was delivered to the mouse mode to investigate the effect of rhuPAa-melittin on the growth of the xenotransplanted tumor. Results rhuPAa-melittin was used to treat ovarian cancer cells at the concentration of 0 ,4 ,8 and 16 μg/mL for 48 hrs ,respectively. The results of cell apoptosis assay was 1.16%,3.83%,6.51% and 10.2%,respectively. Moreover,different concentrations of rhuPAa-melittin had no effects on the cells at G0/G1 phase,rhuPAa-melittin inhibited S phase cells to process into G2/M phase, contributing to suppressing the growth of ovarian cancer cells. Conclusion The in vivo and in vitro studies revealed that rhuPAa-melittin inhibits the growth of ovarian cancer.
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Objective To investigate the role of the activated rhuPAa-melittin in ovarian cancer cells and to study the inhibitory effect of rhuPAa-melittinon on ovarian cancer cells. Methods rhuPAa-melittin was used to treat the ovarian cancer cells at different concentrations for 48 hrs. Then flow cytometery was applied to detect the cell cycle and cell apoptosis. rhuPAa-melittin protein was delivered to the mouse mode to investigate the effect of rhuPAa-melittin on the growth of the xenotransplanted tumor. Results rhuPAa-melittin was used to treat ovarian cancer cells at the concentration of 0 ,4 ,8 and 16 μg/mL for 48 hrs ,respectively. The results of cell apoptosis assay was 1.16%,3.83%,6.51% and 10.2%,respectively. Moreover,different concentrations of rhuPAa-melittin had no effects on the cells at G0/G1 phase,rhuPAa-melittin inhibited S phase cells to process into G2/M phase, contributing to suppressing the growth of ovarian cancer cells. Conclusion The in vivo and in vitro studies revealed that rhuPAa-melittin inhibits the growth of ovarian cancer.