2.MiR-204 inhibits invasion and metastasis of breast cancer cells by targeted regulation of HNRNPA2B1.
Liping ZHANG ; Jun BAI ; Yaqiong HU ; Dandan ZHOU ; Quan ZHENG ; Chonggao YIN ; Qingjie MU ; Hongli LI
Journal of Southern Medical University 2020;40(6):869-875
OBJECTIVE:
To investigate the effect of miR-204 on the invasion and metastasis of breast cancer by targeted regulation of HNRNPA2B1.
METHODS:
The bioinformatics database was used to obtain data of the expressions of miR-204 in breast cancer patients and the survival rate of the patients. RT-qPCR was used to detect the expression of miR-204 in breast cancer cell lines. The expression vector GV369-miR-204 was used to overexpress miR-204 in MDA-MB-231 cells. Transwell assay was performed to detect the effect of miR-204 on the migration and invasion ability of the breast cancer cells. The key genes (hub genes) of miR-204 were determined by bioinformatics method. A dual luciferase assay was used to analyze the targeting relationship between miR-204 and HNRNPA2B1. The expression of HNRNPA2B1 in MDA-MB-231 cells after miR-204 overexpression was detected by Western blotting, and Transwell assay was used to examine the changes in the cell invasion ability.
RESULTS:
The expression of miR-204 was decreased in both breast cancer tissues, and was significantly lower in breast cancer MDA-MB-231 cells than in MCF-10A cells ( < 0.05). The decreased expression of miR-204 was associated with poorer prognosis of breast cancer patients ( < 0.05). Upregulation of miR-204 in MDA-MB-231 cells significantly inhibited the invasion and migration of the cells ( < 0.05). Analysis of the data from the Starbase revealed that the expression of miR-204-5p was negatively correlated with the expression of HNRNPA2B1, and the expression of HNRNPA2B1 was increased in breast cancer patients ( < 0.05) in association with a poorer prognosis of the patients ( < 0.05). Dual luciferase assay demonstrated that miR-204 could bind to HNRNPA2B1 in a target-specific manner. Western blotting and Transwell assay showed that miR-204 significant inhibited the migration and invasion ability of breast cancer cells by targeting HNRNPA2B1 ( < 0.05).
CONCLUSIONS
miR-204 expression is decreased in breast cancer tissues and cells, and its overexpression can inhibit the invasion and metastasis of breast cancer cells by targeted regulation of HNRNPA2B1.
Breast Neoplasms
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Cell Line, Tumor
;
Cell Movement
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Cell Proliferation
;
Gene Expression Regulation, Neoplastic
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Humans
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MicroRNAs
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genetics
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Neoplasm Invasiveness
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Neoplasm Metastasis
3.Mechanism of DERL3 Affecting the Proliferation, Invasion and Metastasis of Lung Adenocarcinoma A549 Cells.
Dandan ZHOU ; Jiemin WANG ; Ke YANG ; Liping ZHANG ; Quan ZHENG ; Jun BAI ; Yaqiong HU ; Qingjie MU ; Chonggao YIN ; Hongli LI
Chinese Journal of Lung Cancer 2020;23(8):638-645
BACKGROUND:
Derlin 3 (DERL3) is downregulated in colorectal cancer (CRC) samples. Its level is closely linked to lymphatic metastasis or distant metastasis rate in CRC patients. However, its biological behavior in lung adenocarcinoma were rarely reported. The aim of this study is to investigate the ectopic expression of DERL3 in lung adenocarcinoma tissues and its effect on the invasion and metastasis of lung adenocarcinoma A549 cell line to reveal the possible mechanism of invasion and metastasis of lung adenocarcinoma.
METHODS:
Lung adenocarcinoma microarray gene chip data included 3 cases of lymph node metastasis and 3 cases of lung adenocarcinoma tissue without lymph node metastasis. The GEDS and Kaplan-Meier plot queries the survival curve and expression level of DERL3. Western blot was used to detect the expression of DERL3 in lung adenocarcinoma cells. The efficiency of knockdown DERL3 gene was detected by Western blot assay. Transwell detected the number of cells passing through the basement membrane of the transwell. EDU assay detected cell proliferation ability. Western blot detected the expression of epithelial-mesenchymal transition related proteins E-cadherin and Vimentin.
RESULTS:
The microarray gene chip results showed that compared with lung adenocarcinoma tissues without lymph node metastasis, 1,314 mRNAs in lung adenocarcinoma tissues with lymph node metastasis were up-regulated, 400 mRNAs were down (P<0.05). The expression of DERL3 increased in lung adenocarcinoma (P<0.05). The results of survival curve showed that the lung cancer patients with high expression of DERL3 with poor prognosis (P<0.05). Western blot results indicated that plasmid transfection was successful. Knockdown of DERL3 suppressed the ability of proliferation, invasion and migration in A549 cells (P<0.05). After knockdown of DERL3, the expression level of Vimentin was decreased, while E-cadherin expression increased (P<0.05).
CONCLUSIONS
Knockdown of DERL3 inhibited the proliferation, invasion and metastasis of A549 cells.
4.CCL18 Promotes the Invasion of Lung Adenocarcinoma through ANXA2.
Zikun DENG ; Qinpei XIAO ; Yuanhang ZHENG ; Ruijun FENG ; Zhimei SHENG ; Baogang ZHANG
Chinese Journal of Lung Cancer 2021;24(7):461-467
BACKGROUND:
ANXA2 plays a very important role in cancer progression. chemokine ligand 18 (CCL18) is associated with the invasion, migration, metastasis and poor prognosis of lung adenocarcinoma (LUAD). In this study, we aimed to explore whether CCL18 promotes LUAD invasion through ANXA2, and its role and molecular mechanism in LUAD invasion.
METHODS:
Western blot was used to detect ANXA2 expression in LUAD tissues and adjacent non-tumor tissues, the transfection efficiency of SiANXA2#2 in cells and the role of ANXA2 as an upstream regulator in the AKT/cofilin signaling pathway. In vitro cytological experiments such as chemotaxis experiment and transwell invasion test was used to explore the mechanism of ANXA2 on LUAD metastasis. F-actin polymerization experiment and Western blot were used to detect whether invasion ability alteration of SiANXA2#2 A549 cells are related to F-actin.
RESULTS:
Western blot analysis showed that compared with adjacent non-tumor tissues, the protein expression level of ANXA2 in cancer tissues increased (P<0.05). In the chemotaxis experiment and invasion experiment, the chemotaxis and invasion ability induced by CCL18 decreased when ANXA2 knockdowned (P<0.05). Compared with the control group, F-actin polymerization was significantly lower in ANXA2 knockdown group, while phosphorylation of AKT at Ser473 and Thr308 and phosphorylation of Cofilin and LIMK were reduced in ANXA2 knockdown group (P<0.05).
CONCLUSIONS
ANXA2 knockdown can reduce the invasive effect of CCL18 on LUAD cells by reducing phosphorylation of AKT and downstream pathways.
5.Differentiation of Monocyte derived Dendritic Cells in Patients with Psoriasis
Kejian ZHU ; Weifang ZHOU ; Limin LAO ; Min ZHENG
Chinese Journal of Dermatology 1994;0(02):-
Objective To investigate the differentiation of dendritic cells from monocytes of the peripheral blood in patients with psoriasis. Methods Flow cytometry was used to analyze the phenotype of monocyte derived dendritic cells (MoDC). The capacity of MoDC to stimulate the proliferation of T lymphocytes was evaluated by allogeneic mixed lymphocyte reaction. Results Monocytes in the peripheral blood of patients with psoriasis could differentiate into dendritic cells. Expression of CD40, CD80, CD86 and HLA-DR by MoDC was significantly increased in patients with psoriasis compared with that in normal controls (P
6.The value of end-tidal carbon dioxide partial pressure combined passive leg raising test on volume responsiveness assessment in shocked patients post cardiac operation
Tao YAO ; Weifang WU ; Xia ZHENG ; Qiang FANG
Chinese Critical Care Medicine 2016;28(5):391-395
Objective To assess the value of end-tidal carbon dioxide partial pressure (PETCO2) combined passive leg raising (PLR) test on volume responsiveness assessment in shocked patients post cardiac operation.Methods A prospective,self-controlled,and observational study was conducted.The shocked patients post cardiac operation undergoing complete mechanical ventilation admitted to Department of Critical Care Medicine of First Affiliated Hospital of College of Medicine,Zhejiang University from June 2014 to October 2015 were enrolled.PETCO2 and hemodynamic parameters including stroke volume variation (SVV),cardiac index (CI),mean arterial pressure (MAP) monitored by a pulse indicator continuous cardiac output (PiCCO) were determined before and after PLR and volume expansion (VE).Volume responsiveness was defined as an increase in CI (△ CI) of 15% or greater after VE,namely response group (△ CI ≥ 15%) and non-response group (△ CI < 15%).The value of PLR-induced PETCO2 change (△PETCO2 PLH) to predict volume responsiveness was evaluated by receiver operating.characteristic curves (ROC).Results Among the 41 patients enrolled,21 had volume responsiveness (response group),and 20 had no responsiveness (non-response group).After PLR,the changes in CI and PETCO2 were both significantly increased in the response group compared with non-response group [△ CI:(13.5 ± 4.6)% vs.(3.6± 3.5)%,△ PETCO2:(7.4 ± 3.4)% vs.(2.8 ± 2.5)%,both P < 0.05].△ PETCO2 PLR and baseline SVV were positively correlated with PLR-induced CI change (△ CI PLR) (r1 =0.50,r2 =0.38,both P < 0.05).VE-induced PETCO2 change (△ PETCO2 VE),baseline SVV and △ CI PLR were positively correlated with VE-induced CI (△ CI VE) (r1 =0.58,r2 =0.56 and r3 =0.84,all P < 0.01).The area under ROC curve (AUC) of △ PETCO2 PLR was 0.875±0.054 [95% confidence interval (95%CI) =0.769-0.981,P < 0.05].△ PETCO2 PLR ≥ 5.8% predicted volume responsiveness with sensitivity of 76.2% and specificity of 90.0%.AUC of △CI PLR was 0.933±0.036 (95%CI =0.862-1.000,P < 0.05).△CI PLR ≥ 10.4% predicted volume responsiveness with sensitivity of 81.0% and specificity of 90.0%.AUC of baseline SVV was 0.831 ±0.066 (95%CI =0.702-0.960,P < 0.05).Baseline SVV ≥ 12.5% predicted volume responsiveness with sensitivity of 85.7% and specificity of 75.0%.Conclusion The change in PETCO2 induced by PLR is a convenient,reliable and non-invasive indicator to predict volume responsiveness in shocked patients post cardiac operation with mechanical ventilation.
8.Effect of Rehabilitation on Psychological Health in First Schizophrenic Patients
Chunxia FANG ; Weifang ZHENG ; Lijun ZHANG ; Jinhong XIA
Chinese Journal of Rehabilitation Theory and Practice 2011;17(1):83-84
ObjectiveTo study the effect of rehabilitation on psychological health in first schizophrenic patients. Methods86 patients were divided into observation group and control group equally. Patients in both groups received medical treatment and routine health education. Patients in observation group received rehabilitation in addition. They were assessed with Symptom Checklist-90 (SCL-90)before and 8 weeks after treatment.ResultsAfter treatment, the scores of SCL-90 in both groups reduced significantly (P<0.05), and less in the observation group than in the control group (P<0.01).ConclusionRehabilitation can improve the psychological health for patients with first schizophrenia.
9.SP13786 Inhibits the Migration and Invasion of Lung Adenocarcinoma Cell A549 by Supressing Stat3-EMT via CAFs Exosomes.
Shushu WANG ; Jiayu CUI ; Kaijia ZHANG ; Jinhua GU ; Yuanhang ZHENG ; Baogang ZHANG ; Lihong SHI
Chinese Journal of Lung Cancer 2021;24(6):384-393
BACKGROUND:
Fibroblast activation protein (FAP) is one of the surface markers of cancer-associated fibroblasts (CAFs) and is closely related to the malignant characterization of CAFs. SP13786 is a specific micromolecule inhibitor of FAP and this study is to investigate the effects and mechanism of SP13786 on the migration and invasion of A549 cells through regulating exosomes of CAFs.
METHODS:
CAFs and paracancerous fibroblasts (PTFs) were isolated and subcultured from freshly resected lung adenocarcinoma tissues and paracancerous normal tissues separately. MTT assay was used to detect the proliferation of CAFs incubated by different concentrations of SP13786; PTFs-exo, CAFs-exo and CAFs+SP13786-exo were extracted by polymer precipitation method. The A549 cells were divided into Ctrl group, PTFs group, CAFs group and SP13786 group and each group was incubated with DMEM, PTFs-exo, CAFs-exo and CAFs+SP13786-exo separately. Laser confocal microscope was used to observe the endocytoses of exosomes by A549 cells. The expression of alpha-smooth muscle actin (α-SMA) and FAP in PTFs and CAFs and the expression of E-cadherin, N-cadherin, Slug, Stat3 and P-Stat3 in A549 cells were detected by immunofluorescence, immunohistochemistry and Western blot. The migration and invasion ability of A549 cells were detected by cell scratch and transwell methods.
RESULTS:
α-SMA and FAP were expressed much higher in CAFs than that in PTFs which indicate that CAFs and PTFs were successfully obtained from lung adenocarcinoma and paracancerous tissues (P<0.05). MTT showed that the 50% inhibitory concentration (IC50) of SP13786 for CAFs was about 3.3 nmol/L. In addition, SP13786 can significantly decrease the expression of α-SMA and FAP in CAFs which means that targeted inhibition of FAP could reduce the malignant characteristics of CAFs (P<0.05). Laser confocal microscope found that exosomes from CAFs could be taken up by A549 cells and scratch and transwell tests showed that the endocytosed CAFs-exo could promote the migration and invasion of A549 cells (P<0.001), while FAP inhibitor SP13786 could inhibit the effects of CAFs-exo on A549 cells (P<0.05). Furthermore, Immunofluorescence and Western blot showed that CAFs-exo could promote EMT by decreasing E-cadherin expression and increasing N-cadherin, Slug expression in A549 cells while FAP inhibitor SP13786 could significantly supress CAFs-exo-induced epithelial-mesenchymal transition (EMT) of A549 cells (P<0.05). Moreover, the expression of P-Stat3 was obviously increased in A549 cells of CAFs group and significantly down-regulated in SP13786 group (P<0.05) whereas there was no significant difference in total Stat3 between CAFs and SP13786 groups (P>0.05). Finally, WP1066 (a specific inhibitor of Stat3) was used to comfirm whether SP13786 could influence EMT of A549 cells by inhibiting Stat3 phosphorylation via CAFs-Exo. The results showed that when the phosphorylation of Stat3 in CAFs group was inhibited by WP1066, SP13786 could not influence the P-Stat3 expression and EMT of A549 cells anymore (P>0.05).
CONCLUSIONS
As a specific micromolecule inhibitor of FAP, SP13786 indirectly inhibits the migration and invasion of A549 cells by affecting exosomes of CAFs. The possible mechanism is to inhibit the phosphorylation of Stat3 and thus affect the EMT of A549 cells.
10.Onset of Coronary Heart Disease is Associated with HCMV Infection and Increased CD14 CD16 Monocytes in a Population of Weifang, China.
Hong Zheng LI ; Qin WANG ; Yi Yuan ZHANG ; Jin Dong WANG ; Hong Juan WU ; Mo Gen ZHANG ; Ji Chen LI ; Zhi Jun LIU
Biomedical and Environmental Sciences 2020;33(8):573-582
Objective:
To investigate the relationship between human cytomegalovirus (HCMV) infection and peripheral blood CD14 CD16 monocytes in the pathogenesis of coronary heart disease (CHD), and to elucidate the mechanism of pathogenesis in CHD by analyzing the correlation between infection, inflammation, and CHD, to provide a basis for the prevention, evaluation, and treatment of the disease.
Methods:
In total, 192 patients with CHD were divided into three groups: latent CHD, angina pectoris, and myocardial infarction. HCMV-IgM and -IgG antibodies were assessed using ELISA; CD14 CD16 monocytes were counted using a five-type automated hematology analyzer; mononuclear cells were assessed using fluorescence-activated cell sorting; and an automatic biochemical analyzer was used to measure the levels of triglyceride, cholesterol, high- and low-density lipoprotein cholesterols, lipoprotein, hs-CRp and Hcy.
Results:
The positive rates of HCMV-IgM and -IgG were significantly higher in the CHD groups than in the control group. HCMV infection affects lipid metabolism to promote immune and inflammatory responses.
Conclusion
HCMV infection has a specific correlation with the occurrence and development of CHD. The expression of CD14 CD16 mononuclear cells in the CHD group was increased accordingly and correlated with acute HCMV infection. Thus, HCMV antibody as well as peripheral blood CD14 CD16 mononuclear cells can be used to monitor the occurrence and development of CHD.
Angina Pectoris
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epidemiology
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virology
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China
;
epidemiology
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Coronary Disease
;
epidemiology
;
virology
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Cytomegalovirus
;
physiology
;
Cytomegalovirus Infections
;
complications
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Humans
;
Incidence
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Inflammation
;
epidemiology
;
etiology
;
Leukocyte Count
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Monocytes
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metabolism
;
Myocardial Infarction
;
epidemiology
;
virology