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OBJECTIVE@#To investigate the effect and potential mechanism of dihydromyricetin (Dmy) on H9C2 cell proliferation, apoptosis, and autophagy.@*METHODS@#H9C2 cells were randomly divided into 7 groups, namely control, model, EV (empty pCDH-CMV-MCS-EF1-CopGFP-T2A-Puro vector), IV (circHIPK3 interference), Dmy (50 µ mol/L), Dmy+IV, and Dmy+EV groups. Cell proliferation and apoptosis were detected by cell counting kit-8 assay and flow cytometry, respectivley. Western blot was used to evaluate the levels of light chain 3 II/I (LC3II/I), phospho-phosphoinositide 3-kinase (p-PI3K), protein kinase B (p-AKT), and phospho-mammalian target of rapamycin (p-mTOR). The level of circHIPK3 was determined using reverse transcriptase polymerase chain reaction. Electron microscopy was used to observe autophagosomes in H9C2 cells.@*RESULTS@#Compared to H9C2 cells, the expression of circHIPK in H9C2 hypoxia model cells increased significantly (P<0.05). Compared to the control group, the cell apoptosis and autophagosomes increased, cell proliferation rate decreased significantly, and the expression of LC3 II/I significantly increased (all P<0.05). Compared to the model group, the rate of apoptosis and autophagosomes in IV, Dmy, and Dmy+IV group decreased, the cell proliferation rate increased, and the expression of LC3 II/I decreased significantly (all P<0.05). Compared to the control group, the expressions of p-PI3K, p-AKT, and p-mTOR in the model group significantly reduced (P<0.05), whereas after treatment with Dmy and sh-circHIPK3, the above situation was reversed (P<0.05).@*CONCLUSION@#Dmy plays a protective role in H9C2 cells by inhibiting circHIPK expression and cell apoptosis and autophagy, and the mechanism may be related to PI3K/AKT/mTOR pathway.
Subject(s)
Proto-Oncogene Proteins c-akt/metabolism , Signal Transduction , Phosphatidylinositol 3-Kinases/metabolism , TOR Serine-Threonine Kinases/metabolism , Apoptosis , AutophagyABSTRACT
Objective:To investigate the mechanism of BCYRN1 regulating miR-503 through Notch1 signaling pathway in the migration and invasion of lung cancer. Methods:The expression of BCYRN1 and miR-503 in different lung cancer cell lines were detected by qPCR. Immunofluorescence and qPCR were used to detect the transfection efficiency of lentivirus BCYRN1 + siRNA transfected lung cancer cells. Double luciferase reporter gene was used to detect the interaction between BCYRN1 and miR-503. Transwell invasion test and scratch test were used to detect the invasion and migration of lung cancer cells after silencing BCYRN1. Western blot was used to detect the expression of Notch1 signaling pathway silence BCYRN1. The effect of silencing BCYRN1 on lung cancer cells in nude mice was measured by subcutaneous tumor formation in nude mice. Results:The expression level of BCYRN1 was the highest in lung cancer cell H1299,and the expression level of miR-503 was relatively high. Immunofluorescence and mRNA levels demonstrated that BCYRN1 + siRNA lentivirus could be effectively transfected into H1299 cells;BCYRN19 binds specifically to the 3′-UTR of miR-503;silencing BCYRN1 could inhibit the invasion and migration of lung cancer H1299 cells;the expression of Notch1 pathway protein was down-regulated after silencing BCYRN1. Compared with NC group,tumor volume and weight of BCYRN1-siRNA group were significantly decreased. Conclusion:BCYRN1 can target the regulation of miR-503 through Notch1 signaling pathway in the invasion and migration of lung cancer H1299 cells.
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Objective:To investigate the mechanism of BCYRN1 regulating miR-503 through Notch1 signaling pathway in the migration and invasion of lung cancer. Methods:The expression of BCYRN1 and miR-503 in different lung cancer cell lines were detected by qPCR. Immunofluorescence and qPCR were used to detect the transfection efficiency of lentivirus BCYRN1 + siRNA transfected lung cancer cells. Double luciferase reporter gene was used to detect the interaction between BCYRN1 and miR-503. Transwell invasion test and scratch test were used to detect the invasion and migration of lung cancer cells after silencing BCYRN1. Western blot was used to detect the expression of Notch1 signaling pathway silence BCYRN1. The effect of silencing BCYRN1 on lung cancer cells in nude mice was measured by subcutaneous tumor formation in nude mice. Results:The expression level of BCYRN1 was the highest in lung cancer cell H1299,and the expression level of miR-503 was relatively high. Immunofluorescence and mRNA levels demonstrated that BCYRN1 + siRNA lentivirus could be effectively transfected into H1299 cells;BCYRN19 binds specifically to the 3′-UTR of miR-503;silencing BCYRN1 could inhibit the invasion and migration of lung cancer H1299 cells;the expression of Notch1 pathway protein was down-regulated after silencing BCYRN1. Compared with NC group,tumor volume and weight of BCYRN1-siRNA group were significantly decreased. Conclusion:BCYRN1 can target the regulation of miR-503 through Notch1 signaling pathway in the invasion and migration of lung cancer H1299 cells.
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Objective To investigate the clinical distribution and epidemiological feature of nine respiratory pathogens of acute respiratory tract infection patients in the northern area of Henan Province,in order to provide reference for clinical effective prevention and treatment of acute respiratory tract infection.Methods The clinical distribution and epidemiological feature of nine respiratory pathogens were analyzed by a retrospective review of 11 135 serum samples which were detected by indirect immunofluorescence method from December 2014 to November 2016.Results There were 2 630 cases with nine respiratory pathogens infection,the infection rate was 23.62％.The main four respiratory pathogens were mycoplasma pneumonia (16.63 ％),influenza B virus (2.41％),parainfluenza virus (2.29 ％),adenovirus (0.79％).The nine respiratory pathogens infection rate of male and female was 21.60％ (1 516/7 020) and 27.07 ％ (1 114/4 115),respectively.The nine respiratory pathogens infection rate of ＜ 1 year group,1-5 years group,6-14 years group,15-30 years group,31-50 years group and ＞ 50 years group was 8.91％ (330/3 702),31.12％ (1 424/4 576),37.36％ (634/1 697),27.10％ (90/332),24.64％ (69/280),15.13 ％ (83/548),respectively.The nine respiratory pathogens infection rate of ＜ 1 year group was lower than that of the other five groups (P ＜ 0.05).The nine respiratory pathogens infection rate of spring group,summer group,autumn group and winter group was 24.66％ (591/2 396) 、23.86％ (631/2 645) 、31.09％ (748/2 406)、17.90％ (660/3 688),respectively.The nine respiratory pathogens infection rate of autumn group was higher than that of the other three groups (P ＜ 0.05).There were 253 patients with polyinfection which all were amphimorphic polyinfection in the 2 630 patients with respiratory pathogens infection,the polyinfection rate was 2.27％.Conclusions Respiratory pathogen infection rate was different because of different gender,different age and different season.The virus was the main pathogen of acute respiratory tract infection.Because of multiplicity of infection and seasonal distribution of pathogens,the prevention of the epidemic should be strengthened in spring and autumn season especially.
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Epigenetics is the study of heritable changes in gene expression other than changes in the underlying DNA sequence. Such changes include DNA methylation, histone modification, chromatin remodeling, genomic imprinting, X chromosome inactivation and non-coding RNA regulation. Recent progresses on epigenetics open new possibilities in tackling these challenging problems in forensic science, including identification of fetal paternity testing in embryonic period, determination of the necessary allele in paternity testing, discrimination of identical twins, origination analysis of micro tissue, verification of forged DNA. This review focuses on epigenetics concept and its latest application in the field of paternity testing, age estimation, discrimination between the twins, identification of tissue of origin, and estimation of postmortem interval.