2.miR-142-5p inhibits the invasion and migration of lung adenocarcinoma H1650 cells by affecting epithelial mesenchymal transformation
WANG Miao ; WANG Yu ; LYU Wei ; DUAN Yuqing ; JIA Yunlong ; LIU Lihua
Chinese Journal of Cancer Biotherapy 2020;27(2):142-148
Objective: To study the expression of miR-142-5p in lung adenocarcinoma tissues, and to explore its effect on proliferation, invasion, migration and epithelieal-mesenchymal transition (EMT) of H1650 cells and the potential mechanisms. Methods:Atotal of 107 pairs of lung adenocarcinoma tissues and corresponding para-cancerous tissues from patients, who underwent tumor resection and were pathologically confirmed at the Department of Thoracic Surgery, the Fourth Hospital of Hebei Medical University between Jan. 2014 and Jan. 2015, were collected for this study; in addition, human lung adenocarcinoma cell lines (H1650, HCC827, A549, H1975, PC9) and human bronchial epithelial BEAS-2B cells were also used in this study. qPCR was used to detect the expression of miR-142-5p in lung adenocarcinoma tissues and cell lines. The correlation between expression of miR-142-5p and clinical features was analyzed.After transfection with miR-142-5p mimics or miR-negative control (miR-NC) plasmid, the proliferation, invasion and migration of H1650 cells were detected with CCK-8, Transwell invasion assay and Wound healing assay, respectively. The bioinforamtics tool was used to predict the target genes of miR-142-5p, and Luciferase reporter gene assay was performed to validate the regulation of miR-142-5p on target gene. Western blotting (WB) was used to detect the expressions of cyclin-dependent kinase 5 (CDK5) and EMTrelated protein. Results: Compared to Para-cancerous tissues and BEAS-2B cells, the expression of miR-142-5p was lower in lung adenocarcinoma tissues and cell lines (all P<0.01). Of the 107 cases of lung adenocarcinoma tissues, 61 cases (57.01%) showed decreased miR-142-5 expression, which was correlated with the TNM stage and lymph node metastasis (both P<0.01). Transfection of miR-142-5p mimics significantly up-regulated the expression of miR-142-5p and decreased the proliferation, invasion and migration of H1650 cells (all P<0.05 or P<0.01). Bioinformatics showed that CDK5 was a target gene of miR-142-5p. Luciferase reporter gene assay and WB validated that miR-142-5p could significantly down-regulate CDK5 expression in H1650 cells, up-regulate the expression of E-cadherin and down-regulate the expressions of N-cadherin, Twist and Snail in H1650 cells (all P<0.01). Conclusion: miR-142-5p is low expressed in lung adenocarcinoma tissues and cell lines; it suppresses the EMT process to inhibit, invasion and migration of H1650 cells via down-regulating the expression of CDK5.
3.Batroxobin reduces intracellular calcium concentration and inhibits proliferation of vascular smooth muscle cells.
Qing-bin SONG ; Min-jie WEI ; Zhi-quan DUAN ; Hai-qiang ZHANG ; L B SCHWARTZ ; Shi-jie XIN
Chinese Medical Journal 2004;117(6):917-921
<b>BACKGROUNDb>Batroxobin (BX), a serine protease used in defibrinogenation and thrombolysis, also has an effect on c-fos gene and growth factor. This study attempted to determine the effects of BX on the proliferation of vascular smooth muscle cells (VSMCs) and calcium metabolism.
<b>METHODSb>VSMCs were treated with BX at concentrations of 0.1, 0.3, or 1.0 mmol/L and cell numbers were determined at 0, 24, 48, and 72 hours. Intracellular calcium concentration ([Ca2+]i) was measured using direct fluorescence methods.
<b>RESULTSb>BX was found to suppress proliferation of VSMCs in a dose-dependent fashion with inhibition rates of 18% and 31% by 48 and 72 hours, respectively. In addition, BX decreases basal [Ca2+]i significantly. The basal level in untreated cells was 162.7 +/- 33.8 nmol/L, and decreased to 131.5 +/- 27.7 nmol/L, 128.3 +/- 28.5 nmol/L, and 125.6 +/- 34.3 nmol/L with the three concentrations of BX, respectively. Noradrenaline (NE)-induced [Ca2+]i stimulation was also attenuated by BX (0.1 mmol/L BX, 20% +/- 8% inhibition; 0.3 mmol/L BX, 54% +/- 11% inhibition; 1.0 mmol/L BX, 62% +/- 15% inhibition). The ability of NE to stimulate [Ca2+]i was attenuated in cultures in Ca(2+)-free medium, as was the ability of BX to blunt NE-induced stimulation.
<b>CONCLUSIONb>These findings demonstrate that BX can effectively inhibit proliferation of VSMCs, probably by blocking the release and uptake of Ca2+, thus influencing [Ca2+]i.
Animals ; Batroxobin ; administration & dosage ; pharmacology ; Calcium ; metabolism ; Cell Division ; drug effects ; Cells, Cultured ; Dose-Response Relationship, Drug ; Muscle, Smooth, Vascular ; cytology ; metabolism ; Rabbits
4.Expression of lncRNA DGCR5 in esophageal squamous cell carcinoma tissues and its clinical significance
DUAN Yuqing ; WANG Mengjie ; WANG Hongyan ; WANG Yu ; SANG Meixiang ; LIU Lihua
Chinese Journal of Cancer Biotherapy 2020;27(4):416-419
[Abstract] Objective: To investigate the expression of long non-coding RNA (lncRNA) DiGeorge syndrome critical region gene 5 (DGCR5) in esophageal squamous cell carcinoma (ESCC) tissues, and to analyze its relationship with clinicopathological features and prognosis of ESCC patients. Methods: The expression of DGCR5 in ESCC data set from TCGA database was analyzed by bioinformatics method. Sixty pairs of ESCC tissues and para-cancerous tissues resected at the Fourth Hospital of Hebei Medical University from August 2016 to March 2017 were collected for this study. The expression of DGCR5 in ESCC tissues was detected by qPCR. The correlation between the expression of DGCR5 and the clinicopathological features and prognosis of ESCC patients was analyzed. Results: TCGAdatabase analysis showed that the expression of DGCR5 in ESCC tissues was significantly higher than that in normal esophageal tissues (P<0.01). The expression of DGCR5 in ESCC tissues was significantly higher than that in para-cancerous tissues (P<0.01). The expression level of DGCR5 was significantly correlated with TNM staging and lymph node metastasis in ESCC patients (all P<0.05). Kaplan-Meier univariate analysis showed that the 2-year survival rate of ESCC patients with high DGCR5 expression was significantly lower than that of patients with low expression (P<0.05). Conclusion: DGCR5 is highly expressed in ESCC tissues and is closely related to TNM staging, lymph node metastasis and poor prognosis, which may serve as a molecular marker for early diagnosis and prognosis prediction of ESCC.
5.Investigation of chigger mites on small mammals in Ruili, Yunnan Province, China
Zhou, Y. ; Duan, B. ; Ren, T.G. ; Dong, W.G.
Tropical Biomedicine 2022;39(No.3):455-461
Chigger mites are arthropods and are the sole vectors of scrub typhus, and rodents as well as other
small mammals are the most common hosts of chigger mite larvae. Therefore, it is of great medical
significance to study the ecology of chigger mites. In this study, a detailed analysis of chigger mites was
conducted based on field survey data. A total of 4,941 chigger mites were collected from 86 hosts at 34
survey sites in Ruili, Yunnan Province, China. Among the 4,941 chiggers, five genera in one subfamily
were identified; Schoengastiella ligula was the dominant chigger species with the highest infestation
index, prevalence (Pm, 42.86%) and mean intensity (MI, 59.09%) (P<0.001). The association coefficient
(V) between S. ligula and Gahrliepia radiopunctata was positively correlated (P<0.05), indicating the
tendency of chiggers to select and coexist on the same host at the same time. The dominant species
Leptotrombidium kunmingense, Ascoschoengastia indica, S. ligula and G. radiopunctata showed
aggregation distribution patterns, indicating that the distribution of chiggers among different hosts
was not uniform. Low altitudes and low latitudes appeared to be more favorable for the growth and
reproduction of chigger mites (P<0.05). It is suggested to collect as many host samples as possible in
future field investigations to better understand the dynamics of chigger mite populations and their
primary hosts.
6.LINC01503 promotes progression of epithelial ovarian cancer through the miR-342-3p/IGF2R axis
LYU Wei1a ; WANG Jiali1a ; LIU Tianxu1a ; DUAN Yuqing1a ; WANG Jun2 ; LIU Lihua1a,1b,1c
Chinese Journal of Cancer Biotherapy 2023;30(9):754-761
[摘 要] 目的:探讨LINC01503在上皮性卵巢癌(EOC)中的表达水平和生物学功能及其可能的作用机制。方法:收集2015年5月至2016年5月间在河北医科大学第四医院妇瘤科手术切除并经病理学确诊的85例EOC患者的肿瘤组织和输卵管组织。常规培养人EOC细胞A2780、SKOV3、OVCAR3和OV90及正常人卵巢上皮细胞IOSE80,将si-LINC01503、si-NC及miR-342-3p mimic、miR mimic NC分别转染至SKOV3和A2780细胞,分别作为si-LINC01503组、si-NC组、miR-342-3p mimic组和miR mimic NC组。qPCR法检测EOC组织和细胞中LINC01503的表达水平,Kaplan-Meier法分析LINC01503表达水平与患者生存的关系。双荧光素酶报告基因实验验证LINC01503/miR-342-3p/IGF2R轴相关分子间的靶向关系。平板克隆、划痕愈合和Transwell实验分别检测敲低LINC01503及转染miR-342-3p mimic对A2780和SKOV3细胞增殖、迁移和侵袭能力的影响。WB法检测EOC细胞中LINC01503/miR-342-3p通路对IGF2R蛋白表达的影响。构建A2780细胞裸鼠移植瘤模型,观察敲低LINC01503对移植瘤生长的影响。结果:EOC组织和细胞中LINC01503表达水平分别显著高于输卵管组织和IOSE80细胞(均P<0.01),LINC01503高表达组患者术后PFS和OS均显著短于LINC01503低表达组患者(均P<0.01)。敲低LINC01503、转染miR-342-3p mimic均可抑制EOC细胞的增殖、迁移和侵袭能力(均P<0.01)。敲低LINC01503可下调IGF2R的表达(P<0.01),这一现象可通过转染miR-342-3p inhibitor挽救。敲低LINC01503可抑制A2780细胞裸鼠移植瘤的生长(P<0.01)。结论:在EOC组织和细胞中呈高表达的LINC01503与患者的不良预后密切相关,LINC01503可能通过吸附miR-342-3p影响IGF2R表达进而促进EOC的进展。
8.HIV gene subtypes of newly reported HIV/AIDS cases in Dehong Dai and Jingpo autonomous prefecture of Yunnan province, 2016.
X DUAN ; K R WANG ; J B WANG ; R H YE ; Y K WANG ; J YANG ; T YANG ; S J ZHOU ; Y C YANG ; S T YAO ; S DUAN ; N HE
Chinese Journal of Epidemiology 2018;39(5):678-681
<b>Objective:b> To explore distribution of HIV gene subtypes among newly reported HIV/AIDS cases from China and Myanmar in Dehong Dai and Jingpo prefecture of Yunnan province in 2016. <b>Methods:b> We conducted DNA extractions from newly reported HIV/AIDS cases in 2016. The gag, env and pol genes were amplified by using reverse transcription-PCR (RT-PCR) and sequenced to identify HIV subtypes. <b>Results:b> A total of 1 112 newly diagnosed HIV cases were reported in Dehong in 2016, and the HIV subtypes were identified for 860 cases. Subtype C was predominant (33.6%), followed by unique recombinant forms (URFs) (28.4%), CRF01_AE (18.6%) and so on. URFs include four recombination, among which the recombination of CRF01_AE and C subtype were predominant. The HIV subtype distribution was associated with nationality and transmission route in HIV/AIDS cases from Myanmar. <b>Conclusions:b> The gene subtypes of C, URFs and CRF01_AE were mainly distributed; distribution of URFs remained complex and diverse among newly reported HIV/AIDS cases in Dehong in 2016.
Base Sequence
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China/epidemiology*
;
Ethnicity/genetics*
;
Genes, pol
;
Genotype
;
HIV Infections/genetics*
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HIV-1/genetics*
;
Humans
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Male
;
Phylogeny
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Polymerase Chain Reaction/methods*
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Serogroup
9.Molecular transmission clusters on HCV genotypes among newly reported HIV/HCV co-infection in Dehong Dai and Jingpo autonomous prefecture of Yunnan province, 2016.
Y K WANG ; X C CHEN ; J B WANG ; X DUAN ; S J ZHOU ; J YANG ; T YANG ; R H YE ; Y C YANG ; S T YAO ; S DUAN ; N HE
Chinese Journal of Epidemiology 2019;40(2):191-195
<b>Objective:b> To understand the characteristics on major strain subtypes of hepatitis C virus among HIV/HCV co-infected patients, so as to explore the molecular transmission clusters and related risk factors of HCV strains. <b>Methods:b> A total of 336 newly reported HIV-infected patients were diagnosed as HIV/HCV co-infection in Dehong Dai and Jingpo autonomous prefecture (Dehong) in 2016. We used Nested PCR to amplify CE1 and NS5B genes among 318 samples with plasma levels above 200 μl, before using the combining phylogenetic tree and constructing molecular propagation network method to analyze the related data. <b>Results:b> A total of 267 HIV/HCV co-infection patients who had met the HCV genotyping requirements were screened the gene subtypes were diversified. Among these genotypes, proportions of 3b, 6n, 6u, 1a, 3a and other subtypes appeared as 32.6% (87/267), 18.4% (49/267), 15.7%(42/267), 13.1%(35/267), 11.2%(30/267) and 9.0%(24/267) respectively. Molecular transmission network of five major HCV genotypes was constructed with a clustering rate of 39.1% (95/243). The clustering rate of subtype 1a was the highest, as 71.4% (25/35). Results from the multivariate logistic regression showed that ethnic minorities other than the Yi and Jingpo (vs. the Han, OR=0.17, 95%CI: 0.04-0.71), the married spouses (vs. the unmarried, OR=0.42, 95%CI: 0.18-0.94), the 6n and 3a subtype (vs. the 3b subtype, OR=0.34, 95%CI: 0.12-0.95; OR=0.22, 95%CI: 0.05-0.93) were more difficult to form transmission clusters. However, the 6u and 1a subtype (vs. the 3b subtype, OR=3.10, 95%CI: 1.21-7.94; OR=4.00, 95%CI: 1.32-12.11) seemed more likely to form the transmission clusters. <b>Conclusion:b> Ethnicity, marital status and genetic subtypes were factors significantly associated with the formation of transmission clusters related to the major HCV gene subtypes among newly reported HIV/HCV co-infection in Dehong.
AIDS-Related Opportunistic Infections/virology*
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Asian People
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China/epidemiology*
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Coinfection
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Genotype
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HIV Infections/virology*
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Hepacivirus/isolation & purification*
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Hepatitis C/virology*
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Humans
;
Phylogeny
;
Polymerase Chain Reaction
10.Identification of metabolites of Zhali Nusi Prescription in rat plasma, bile, urine and feces after oral administration.
Ting ZHANG ; Yang NIU ; Kai-Di HUANG ; B U FAN-SHU ; Xiao-Kun BIAN ; Qiu-Long ZHAO ; Sheng GUO ; Er-Xin SHANG ; Da-Wei QIAN ; Jin-Ao DUAN
China Journal of Chinese Materia Medica 2020;45(21):5280-5288
This study was designed to determine the metabolites of Zhali Nusi Prescription(ZLNSP) in rats. The ultra-high performance liquid chromatography-LTQ Orbitrap mass spectrometric(UHPLC-LTQ-Orbitrap-MS) and mass defect filter techniques were applied to analyze the metabolites of ZLNSP in rat plasma, bile, urine and feces. The biological samples were analyzed by ACQUITY UPLC BEH T_3 column(2.1 mm×100 mm,1.7 μm), with 0.1% formic acid water(A)-acetonitrile(B) as mobile phase, and the biological samples were analyzed in negative ion mode by electrospray ionization mass spectrometry(ESI-MS). An analytical method for biological samples of rats was established, and 8 prototype components and 36 metabolites were identified. The results showed that the metabolic pathways of the main components of ZLNSP in rats included methylation, glucuronidation, sulfation and so on. It provi-ded information for the therapeutic effect of ZLNSP in vivo.
Administration, Oral
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Animals
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Bile
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Chromatography, High Pressure Liquid
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Feces
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Plasma
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Prescriptions
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Rats