1.Activation of liver X receptors induced pancreatic β cell cycle arrest by up-regulating the expression of p27 protein
Xuhua MAO ; Junming TANG ; Guohong QIAO ; Siyi FENG ; Xiao HAN ; Changwen JING
Chinese Journal of Clinical Laboratory Science 2017;35(5):386-389
Objective To investigate the effects of liver X receptor (LXR) agonist on the proliferation of mouse pancreatic β cell line MIN6 cells.Methods The viability,changes of cell cycle,mRNA levels of S phase kinase associated protein 2 (Skp2) and p27,and protein levels of Skp2 and p27 in MIN6 cells treated with LXR agonist T0901317 were determined by the CCK-8 method,flow cytometry,real-time RT-PCR and western blot,respectively.Results The viability of MIN6 cells treated with 1 μmol/L,5 μmol/L and 10 μnol/L of T0901317 were (98.54 ±0.94)%,(87.03 ±0.93)% and (75.57 ± 1.85)% of the controls,respectively,and there was significant difference among them (F =301.90,P < 0.01).The percentages of G1 phase cells in the MIN6 cells treated with 0 μmol/L,1 μmol/L,5 μmol/L and 10 μmol/L of T0901317 were (35.93 ±2.25)%,(38.45 ±0.91)%,(45.46±1.34)% and (53.28 ± 1.14) %,respectively,and there was significant difference among them (F =80.83,P < 0.01).Similarly,the percentages of S phase cells in the MIN6 cells treated with 0 μmol/L,1 μmol/L,5 μmol/L and 10 μmoi/L of T0901317 were (52.87 ± 1.19) %,(48.65 ± 0.85) %,(36.31 ± 1.37) % and (31.45 ± 1.22) %,respectively,and there was also significant difference among them (F =221.30,P < 0.01).The protein levels of p27 in the MIN6 cells treated with 10 μmol/L of T0901317 (2.84 ± 0.14) were significantly higher than that in the controls (2.28 ± 0.10) (t =4.54,P < 0.05),while there was no significant difference in the mRNA levels of p27 between them (t =0.28,P > 0.05).However,10 μmol/L of T0901317 significantly decreased mRNA (0.52 ± 0.02,t =29.22,P < 0.01) and protein levels (0.98 ± 0.12 vs 1.89 ± 0.01,t =10.98,P < 0.01) of Skp2 in MIN6 cells.Based on the control siRNA transfection group as a reference (100%),the cell survival rates of the p27 siRNA transfection group,10 μmol/L of T0901317 treatment group and the intervention group (p27 siRNA transfection + T0901317 treatment) were (100.97 ± 1.08) %,(75.03 ± 1.83) % and (86.67 ± 2.45) %,respectively.There was no significant difference between the control siRNA and p27 siR-NA transfection groups (t =1.542,P > 0.05).Compared with the control siRNA transfection group,the cell survival rates of the T0901317 treatment group decreased (t =23.58,P < 0.01).There was also significant difference in the cell survival rates between the T0901317 treatment group and the intervention group (t =7.77,P < 0.01).Conclusion The activation of LXR may induce pancreatic β cell cycle arrest by up-regulating the expression of p27 and down-regulating the expression of Skp2.
2.Disease modifying therapy of pediatric neuromuscular diseases
Chinese Pediatric Emergency Medicine 2023;30(6):405-410
Neuromuscular diseases (NMDs) are a group of hereditary, commonly childhood-onset neurological disorders affecting various components of motor unit, which are often characterized by reduced muscle strength.With the rapid development of precision medicine, great progress has been made in the pharmacological treatment of NMDs, and pediatric NMDs has also entered the era of disease modifying therapy.Antisense oligonucleotides, gene therapy, small molecule compounds, monoclonal antibodies and other disease modifying therapy drugs targeting the cause and pathogenesis of the diseases are gradually applied in clinical practice.What′s more, a variety of potential new drugs and therapeutic biological products are in continuous development.Hopefully, new breakthroughs are expected to be achieved in the field of pediatric NMDs treatment in the near future.
3.Research progress in the correlation between reproductive tract microbiota and intrauterine adhesion.
Zitong ZHAO ; Xuetao MAO ; Yi ZHENG ; Ying LIU ; Siyi ZHAO ; Shuoyi YAO ; Dabao XU ; Xingping ZHAO
Journal of Central South University(Medical Sciences) 2022;47(11):1495-1503
Intrauterine adhesion (IUA) is caused by damage of the basal layer of endometrium, which leads to fibrosis of the endometrium and the formation of adhesion, resulting in partial or complete occlusion of the uterine cavity, abnormal menstruation, infertility or recurrent miscarriage. The prevalence of IUA in women has been increasing in recent years, and the high recurrence rate of moderate to severe IUA makes IUA treatment more challenging. Iatrogenic endometrial injury is the main cause of IUA. However, the incidence of IUA and the severity of IUA vary among patients who have received similar uterine operations, suggesting that there may be other synergistic factors in the development of IUA. There is a certain correlation between the pathogenesis and the microbiota of the gential tract. In many IUA patients, it has been observed that the probiotics such as Lactobacillus in the vagina is significant reduced, and the pathogenic bacteria such as Gardnerella and Prevotella are excessive growth. The reproductive tract microbiota can be involved in the development and progression of IUA via impacting immune function and metabolism.
Humans
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Female
4.Knockdown of IGF2BP2 inhibits colorectal cancer cell proliferation, migration and promotes tumor immunity by down-regulating MYC expression.
Tianyue LIU ; Chenying HAN ; Chenchen HU ; Siyi MAO ; Yuanjie SUN ; Shuya YANG ; Kun YANG
Chinese Journal of Cellular and Molecular Immunology 2023;39(4):303-310
Objective To investigate the effect of insulin-like growth factor 2 mRNA binding protein 2 (IGF2BP2) on the proliferation, migration and tumor immune microenvironment of colorectal cancer cells and its possible molecular mechanism. Methods The Cancer Genome Atlas (TCGA) database was used to analyze the expression levels of IGF2BP2 and MYC in colorectal cancer and adjacent tissues. The expression of IGF2BP2 in HCT-116 and SW480 human colorectal cancer cells was silenced by RNA interference (RNAi), and the silencing effect was detected by quantitative real-time PCR. After knocking down IGF2BP2, colony formation assay, CCK-8 assay and 5-ethynyl-2'-deoxyuridine (EdU) assay were employed to detect cell colony formation and proliferation ability. TranswellTM assay was used to detect cell migration ability. Quantitative real-time PCR was used to detect the mRNA expression of IGF2BP2, MYC, tumor necrosis factor-α (TNF-α), transforming growth factor-β (TGF-β) and interleukin-10 (IL-10). The protein expression of IGF2BP2 and MYC was detected by western blot. The binding ability of IGF2BP2 and MYC in HCT-116 cells was detected by quantitative real-time PCR after RNA immunoprecipitation. Results The results of TCGA database showed that the expression of IGF2BP2 and MYC in colorectal cancer tissues was significantly higher than that in adjacent tissues, and the survival time of colorectal cancer patients with high expression of IGF2BP2 was shorter. After silencing IGF2BP2, the viability, proliferation and migration of HCT-116 and SW480 cells were decreased. The mRNA expression of MYC, TGF-β and IL-10 in IGF2BP2 knockdown group was significantly decreased, while the expression of TNF-α mRNA was increased. The expression of MYC protein and the stability of MYC mRNA were significantly decreased. RIP-qPCR results showed that IGF2BP2 could bind to MYC mRNA. Conclusion Knockdown of IGF2BP2 inhibits colorectal cancer cell proliferation, migration and promotes tumor immunity by down-regulating MYC expression.
Humans
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Cell Line, Tumor
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Cell Movement/genetics*
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Cell Proliferation/genetics*
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Colorectal Neoplasms/metabolism*
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Gene Expression Regulation, Neoplastic
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Interleukin-10/metabolism*
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RNA, Messenger
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RNA-Binding Proteins/metabolism*
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Transforming Growth Factor beta/genetics*
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Tumor Microenvironment/immunology*
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Tumor Necrosis Factor-alpha/metabolism*
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Proto-Oncogene Proteins c-myc/metabolism*