1.Effect and mechanisms of highly active umbilical cord mesenchymal stem cells on aging spleen in elderly tree shrews
Li YE ; Chuan TIAN ; Xiaojuan ZHAO ; Mengdie CHEN ; Qianqian YE ; Qiang LI ; Zhuyin LIAO ; Ye LI ; Xiangqing ZHU ; Guangping RUAN ; Zhixu HE ; Liping SHU ; Xinghua PAN
Chinese Journal of Tissue Engineering Research 2025;29(19):4000-4010
BACKGROUND:Spleen has the functions of blood storage,hematopoiesis,and immunity.With the increase of age,the structural degeneration and functional decline of spleen lead to the impairment of immune system function,thus accelerating the aging process of the body.The treatment of spleen aging in tree shrews with highly active umbilical cord mesenchymal stem cells has not been reported. OBJECTIVE:To explore the intervention effect and mechanism of highly active umbilical cord mesenchymal stem cells on spleen aging in tree shrews. METHODS:Highly active umbilical cord mesenchymal stem cells were isolated,cultured,and obtained from the umbilical cord tissue of newborn tree shrews by caesarean section.The differentiation abilities of adipogenesis,osteogenesis,and chondrogenesis were detected by three-line differentiation kit.Cell cycle and surface markers were detected by flow cytometry.The second generation of highly active umbilical cord mesenchymal stem cells were transfected with Genechem Green Fluorescent Protein with infection complex values of 100,120,140,160,180,and 200,respectively,to screen the best transfection conditions.After transfection,the fourth generation of highly active umbilical cord mesenchymal stem cells was injected into the tail vein of tree shrews in the elderly treatment group.The young control group and the aged model group were not given special treatment.After 4 months of treatment,the spleen tissue was taken and the structure of the spleen was observed by hematoxylin-eosin staining.β-Galactosidase staining was used to detect the activity of aging-related galactosidase.Immunohistochemical staining was used to detect the expression levels of p21 and p53 proteins.Ki67 and PCNA immunofluorescence staining was used to detect cell proliferation activity.Immunofluorescence staining was used to detect the expression levels of spleen autophagy protein molecules Beclin 1 and APG5L/ATG5.Reactive oxygen species fluorescence staining was used to detect the content of reactive oxygen species in spleen tissue.CD3 immunofluorescence staining was used to detect the change of the proportion of total T lymphocytes.The secretion levels of interleukin 1β and transforming growth factor β1 in spleen were detected by enzyme linked immunosorbent assay.The distribution of highly active umbilical cord mesenchymal stem cells labeled with green fluorescent protein in spleen tissue was observed by DAPI double staining of nucleus. RESULTS AND CONCLUSION:(1)Highly active umbilical cord mesenchymal stem cells grew in a short spindle shape with fish-like growth,with a large proportion of G0/G1 phase,and had the potential to differentiate into adipogenesis,osteogenesis,and chondrogenesis.(2)Multiplicity of infection=140 and transfection for 72 hours were the best conditions for labeling tree shrews highly active umbilical cord mesenchymal stem cells with Genechem Green Fluorescent Protein.(3)Compared with the aged model group,in the aged treatment group,the spleen tissue cells of tree shrews were arranged closely,and the area of white pulp was increased(P<0.01);the boundary between red pulp and white pulp was clear;the proportion of germinal centers did not show statistically significant difference(P>0.05).The activity level of galactosidase related to spleen tissue aging was decreased(P<0.001),and the expression levels of aging protein molecules p21 and p53 were down-regulated(P<0.001).The expression levels of proliferation-related molecules Ki67 and PCNA were up-regulated(P<0.001,P<0.05);expression levels of autophagy-related molecules Beclin 1 and APG5L/ATG5 were up-regulated(P<0.001),and the content of reactive oxygen species decreased(P<0.001),and the proportion of CD3+T cells increased(P<0.05).The secretion level of interleukin 1β in the aging-related secretion phenotype decreased(P<0.001);no significant difference was found in transforming growth factor β1 level(P>0.05).Compared with the young control group,the above indexes were significantly different in the elderly treatment group(P<0.05).(4)Green fluorescent cells labeled with green fluorescent protein were observed in spleen tissue of tree shrews the elderly treatment group by frozen tissue section observation.The results show that intravenous infusion of highly active umbilical cord mesenchymal stem cells can migrate to spleen tissue,inhibit the production of reactive oxygen species,down-regulate the expression of aging-related proteins,induce autophagy,promote cell proliferation,reduce chronic inflammation,and then improve the structure and function of spleen tissue.
2.Incidence of pulmonary tuberculosis and its influencing factors in Hubei Province based on the geographically weighted regression model
Xingxing LU ; Xun LIU ; Fan WANG ; Jianjun YE ; Yu ZHANG ; Chengfeng YANG ; Liping ZHOU ; Hongxing WANG ; Wenqian ZHOU
Journal of Public Health and Preventive Medicine 2025;36(5):28-31
Objective To study the spatial distribution of the incidence of pulmonary tuberculosis in Hubei Province and its influencing factors, so as to improve the theoretical basis for scientific development of tuberculosis prevention and control measures in the future. Methods The data of reported incidence of tuberculosis and related influencing factors in various counties and districts of Hubei Province in 2020 were collected. Global Moran's I index, hotspot analysis and geographically weighted regression (GWR) model analysis were used to calculate the spatial autocorrelation of the incidence of tuberculosis, and to analyze the influencing factors affecting the incidence rate of tuberculosis. Results There were obvious regional differences in the space distribution of the incidence rate of tuberculosis. Hot spot analysis showed positive spatial correlation and obvious clustering. The GWR model (AICc=784.251) in this study had higher AICc value compared to the ordinary least squares regression (OLS) model (AICc=804.2585). The GWR model showed that the increase in the proportion of the population aged 65 and above and the proportion of the ethnic minority population had a significant promoting effect on the increase of the incidence rate of tuberculosis, and there was significant spatial heterogeneity. The effect of PM2.5 concentration on the incidence rate of pulmonary tuberculosis varied in different regions, and the degree of effect was also different. Conclusion The proportion of people aged 65 and above and the proportion of ethnic minorities may significantly influence the incidence of pulmonary tuberculosis. The effect of PM2.5 concentration varies in different regions, so targeted measures should be formulated according to the situation in different regions.
3.Application of P1 response threshold of cortical auditory evoked potential in rehabilitation evaluation of young children with cochlear implant.
Hui JI ; Yaofeng JIANG ; Fei ZHONG ; Baona LI ; Ye FAN ; Shiyu TAO ; Liping MENG
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2025;39(10):962-966
Objective:To explore the application value of P1 response threshold of cortical auditory evoked potential(CAEP) in evaluating the rehabilitation effect of cochlear implant in young children. Methods:Thirty-three young children after cochlear implantation were divided into groups according to hearing age: Group A(hearing age 1-<2 years old) 10 people; Group B(hearing age 2-<3 years old) 13 people; Group C(hearing age 3-<4 years old) 10 people. The subjective assessment was carried out using the assessment tool for hearing-impaired children- "Criteria and Methods for assessing Auditory and language ability of hearing-impaired children" and objective electrophysiological examination was carried out using CAEP to evaluate the rehabilitation effect. SPSS 25.0 software was used for statistical analysis. Results:The results of subjective assessment of auditory ability and language ability in each group showed an increasing trend with the increase of auditory age. In this study, the P1 response threshold of CAEP in CI implanted children had a significant positive correlation with the 2 kHz hearing threshold after intervention, and the P1 response threshold of CAEP was negatively correlated with many items in subjective auditory ability and language ability assessment. Conclusion:The P1 response threshold of CAEP has a stable correlation with the results of speech audiometry, which can effectively and objectively evaluate the postoperative rehabilitation effect of young children with cochlear implantation.
Humans
;
Child, Preschool
;
Infant
;
Male
;
Female
;
Evoked Potentials, Auditory
;
Cochlear Implantation/rehabilitation*
;
Cochlear Implants
;
Auditory Threshold
4.Yiqi Yangyin Huazhuo Tongluo Formula alleviates diabetic podocyte injury by regulating miR-21a-5p/FoxO1/PINK1-mediated mitochondrial autophagy.
Kelei GUO ; Yingli LI ; Chenguang XUAN ; Zijun HOU ; Songshan YE ; Linyun LI ; Liping CHEN ; Li HAN ; Hua BIAN
Journal of Southern Medical University 2025;45(1):27-34
OBJECTIVES:
To investigate the protective effect of Yiqi Yangyin Huazhuo Tongluo Formula (YYHT) against high glucose-induced injury in mouse renal podocytes (MPC5 cells) and the possible mechanism.
METHODS:
Adult Wistar rats were treated with 19, 38, and 76 g/kg YYHT or saline via gavage for 7 days to prepare YYHT-medicated or blank sera for treatment of MPC5 cells cultured in high glucose (30 mmol/L) prior to transfection with a miR-21a-5p inhibitor or a miR-21a-5p mimic. The changes in miR-21a-5p expressions and the mRNA levels of FoxO1, PINK1, and Parkin in the treated cells were detected with qRT-PCR, and the protein levels of nephrin, podocin, FoxO1, PINK1, and Parkin were detected with Western blotting. Autophagic activity in the cells were evaluated with MDC staining. The effect of miR-21a-5p mimic on FoxO1 transcription and the binding of miR-21a-5p to FoxO1 were examined with luciferase reporter gene assay and radioimmunoprecipitation assay.
RESULTS:
MPC5 cells exposed to high glucose showed significantly increased miR-21a-5p expression, lowered expressions of FoxO1, PINK1, and Parkin1 mRNAs, and reduced levels of FoxO1, PINK1, parkin, nephrin, and podocin proteins and autophagic activity. Treatment of the exposed cells with YYHT-medicated sera and miR-21a-5p inhibitor both significantly enhanced the protein expressions of nephrin and podocin, inhibited the expression of miR-21a-5p, increased the mRNA and protein expressions of FoxO1, PINK1 and Parkin, and upregulated autophagic activity of the cells. Transfection with miR-21a-5p mimic effectively inhibited the transcription of FoxO1 and promoted the binding of miR-21a-5p to FoxO1 in MPC5 cells, and these effects were obviously attenuated by treatment with YYHT-medicated sera.
CONCLUSIONS
YYHT-medicated sera alleviate high glucose-induced injury in MPC5 cells by regulating miR-21a-5p/FoxO1/PINK1-mediated mitochondrial autophagy.
Animals
;
MicroRNAs/genetics*
;
Podocytes/pathology*
;
Drugs, Chinese Herbal/pharmacology*
;
Autophagy/drug effects*
;
Rats, Wistar
;
Protein Kinases/metabolism*
;
Rats
;
Forkhead Box Protein O1
;
Mice
;
Mitochondria/drug effects*
;
Ubiquitin-Protein Ligases/metabolism*
;
Glucose
;
Diabetic Nephropathies
;
Male
;
Membrane Proteins/metabolism*
;
Intracellular Signaling Peptides and Proteins
5.Effect of KIAA1522 on proliferation,migration,and invasion of lung cancer cells and its mechanism
Yihui WANG ; Qing ZHANG ; Yingnan LI ; Liping YE
Journal of Jilin University(Medicine Edition) 2025;51(3):727-739
Objective:To discuss the effect of KIAA1522 on the proliferation,migration,and invasion of lung cancer cells,and to clarify its signaling mechanism.Methods:Bioinformatics analysis was used to detect the expression levels of KIAA1522 mRNA and protein in 75 cases of human non-small cell lung cancer(NSCLC)tissues and adjacent normal lung tissues;immunohistochemical staining was used to detect the expression of KIAA1522 protein in NSCLC tissue and adjacent normal lung tissues;Western blotting method was used to detect the expression level of KIAA1522 protein in various lung cancer cell lines.KIAA1522-small interfering(siRNA)and over-expression plasmids were transfected into the lung cancer H1299 and A549 cells,respectively.The KIAA1522-siRNA experiment was divided into blank group,negative control group(si-NC group),KIAA1522-siRNA#1 group,and KIAA1522-siRNA#2 group.The KIAA1522 over-expression experiment was divided into control group,empty vector control group(OE-NC group,transfected with KIAA1522 over-expression empty vector plasmid),KIAA1522 overexpression group(OE-KIAA1522 group,transfected with KIAA1522 over-expression plasmid),KIAA1522 over-expression+MK2206 group[OE-KIAA1522+MK2206 group,co-transfected with KIAA1522 over-expression plasmid and protein kinase B(AKT)signaling pathway inhibitor MK2206],and MK2206 group(transfected with MK2206).Western blotting method was used to verify the transfection efficiencies of the cells in various groups;MTT assay was used to detect the proliferation activities of the lung cancer cells in various groups;cell scratch assay was used to detect the migration rates of lung cancer cells in various groups;Transwell chamber assay was used to detect the numbers of invasion lung cancer cells in various groups;Western blotting method was used to detect the expression levels of phosphorylated AKT(p-AKT),total AKT(t-AKT),cyclin D1(Cyclin D1),vascular endothelial growth factor(VEGF),and epithelial-mesenchymal transition(EMT)-related proteins[vimentin(Vimentin),N-cadherin(N-cadherin),and E-cadherin(E-cadherin)]proteins in the cells in various groups.Results:The bioinformatics analysis results showed that compared with adjacent normal lung tissue,the expression levels of KIAA1522 mRNA and protein in NSCLC tissue were significantly increased(P<0.05 or P<0.01).The immunohistochemistry staining results showed that compared with adjacent normal lung tissue,the positive expression rate of KIAA1522 protein in NSCLC tissue was significantly increased(P<0.05)and was associated with TNM stage(P<0.01).The Western blotting results showed that compared with normal lung epithelial cells BEAS-2B,the expression levels of KIAA1522 protein in lung cancer cell lines PC9,H1299,H460,A549,H1975,and H226 were significantly increased(P<0.05 or P<0.01).Compared with si-NC group,the expression levels of KIAA1522 protein in the H1299 cells in KIAA1522-siRNA#1 group and KIAA1522-siRNA#2 group were significantly decreased(P<0.01);compared with OE-NC group,the expression level of KIAA1522 protein in the A549 cells in OE-KIAA1522 group was significantly increased(P<0.01).The MTT results showed that at 24,48,and 72 h of cell culture,compared with si-NC group,the proliferation activities of the H1299 cells in KIAA1522-siRNA#1 group and KIAA1522-siRNA#2 group were significantly decreased(P<0.01);compared with OE-NC group,the proliferation activity of the A549 cells in OE-KIAA1522 group was significantly increased(P<0.05);compared with OE-KIAA1522 group,the proliferation activity of the A549 cells in OE-KIAA1522+MK2206 group was significantly decreased(P<0.01);compared with OE-KIAA1522+MK2206 group,the proliferation activity of the A549 cells in MK2206 group was significantly decreased(P<0.05).The cell scratch assay results showed that compared with si-NC group,the migration rates of the H1299 cells in KIAA1522-siRNA#1 group and KIAA1522-siRNA#2 group were significantly decreased(P<0.01);compared with OE-NC group,the migration rate of the A549 cells in OE-KIAA1522 group was significantly increased(P<0.01);compared with OE-KIAA1522 group,the migration rate of the A549 cells in OE-KIAA1522+MK2206 group was significantly decreased(P<0.05);compared with OE-KIAA1522+MK2206 group,the migration rate of the A549 cells in MK2206 group was significantly decreased(P<0.05).The Transwell chamber assay results showed that compared with si-NC group,the numbers of invasion H1299 cells in KIAA1522-siRNA#1 group and KIAA1522-siRNA#2 group were significantly decreased(P<0.01);compared with OE-NC group,the number of invasion A549 cells in OE-KIAA1522 group was significantly increased(P<0.01);compared with OE-KIAA1522 group,the number of invasion A549 cells in OE-KIAA1522+MK2206 group was significantly decreased(P<0.01);compared with OE-KIAA1522+MK2206 group,the number of invasion A549 cells in MK2206 group was significantly decreased(P<0.01).The Western blotting results showed that compared with si-NC group,the expression levels of p-AKT,Cyclin D1,Vimentin,N-cadherin,and VEGF proteins in the H1299 cells in KIAA1522-siRNA#1 group and KIAA1522-siRNA#2 group were significantly decreased(P<0.05 or P<0.01),while the expression level of E-cadherin protein was significantly increased(P<0.01);compared with OE-NC group,the expression levels of p-AKT,Cyclin D1,Vimentin,N-cadherin,and VEGF proteins in the A549 cells in OE-KIAA1522 group were significantly increased(P<0.05 or P<0.01),while the expression level of E-cadherin protein was significantly decreased(P<0.05);compared with OE-KIAA1522 group,the expression levels of p-AKT,Cyclin D1,Vimentin,N-cadherin,and VEGF proteins in the A549 cells in OE-KIAA1522+MK2206 group were significantly decreased(P<0.05 or P<0.01),while the expression level of E-cadherin protein was significantly increased(P<0.05);compared with OE-KIAA1522+MK2206 group,the expression levels of Cyclin D1,Vimentin,N-cadherin,and VEGF proteins in the A549 cells in MK2206 group were significantly decreased(P<0.05 or P<0.01),while the expression level of E-cadherin protein was significantly increased(P<0.05).Conclusion:The KIAA1522 protein upregulates the expression of Cyclin D1,EMT-related proteins,and VEGF protein in lung cancer cells,promoting the proliferation,migration,and invasion of lung cancer cells,and its mechanism is related to the activation of the AKT signaling pathway.
6.Characteristics of 456 severe acute respiratory infection cases in Pudong New Area, Shanghai, 2023
Zou CHEN ; Liping CHAI ; Yifeng SHEN ; Chuchu YE ; Yuanping WANG ; Bing ZHAO ; Qi ZHAO ; Hong ZHANG ; Lipeng HAO
Shanghai Journal of Preventive Medicine 2025;37(12):1044-1047
ObjectiveTo retrospectively analyze the association between novel coronavirus (“SARS-CoV-2”) infection and clinical symptoms in inpatients with severe acute respiratory infection (SARI) in Pudong New Area, Shanghai, so as to provide evidence for improving clinical diagnostic ability. MethodsFrom January 13 to March 2, 2023, respiratory tract specimens of 456 inpatients with SARI were collected from 8 sentinel institutions, SARS-CoV-2 was detected by real-time fluorescence quantitative PCR. Whole genome sequencing and sequence analyses were performed to samples with a cycle threshold (Ct) value of <35. At the same time, demographic information, clinical characteristics and underlying disease condition of the cases were collected, and the association between age, symptoms and nucleic acid positive rates was evaluated by χ2 test and Spearman correlation analysis. ResultsA total of 456 cases were included, the median (P25, P75) age was 70 (69, 85) years old, of which 200 cases were novel coronavirus nucleic acid positive for SARS-CoV-2, with a positive rate of 43.86%. The positive rate was the highest in the 80-year-old group (56.82%), and the positive rate increased significantly with age (r=0.15, P=0.002). The proportion of oppression in chest, sore throat and expectoration in novel coronavirus nucleic acid positive cases was significantly higher than that in negative cases (all P<0.05). The 33 viruses sequenced successfully were all Omicron subvariants, with BF.7 (51.52%) and BA.5.2 (42.42%) being the predominant ones. ConclusionA positive correlation was observed between advanced age and the risk of SARS-CoV-2 positivity in patients with SARI. The symptoms of expectoration, oppression in chest and sore throat are more common in positive cases, which can be used as a prompt indicator for key screening and clinical identification of elderly SARI cases.
7.Analysis of Clinical Characteristics in 2 Cases of Hypoparathyroidism Sensorineural Deafness and Renal Dysplasia Syndrome
Min LIU ; Liping MENG ; Hui JI ; Ye FAN ; Yingying WANG ; Qin HONG
Journal of Audiology and Speech Pathology 2024;32(5):422-426
Objective To investigate the clinical characteristics and genetic causes in 2 patients with hypopar-athyroidism,sensorineural deafness and renal dysplasia syndrome(HDR).Methods A retrospective analysis of au-diology,gene detection,and other clinical diagnostic data was performed on 2 patients diagnosed with HDR syn-drome.Results Patient 1 failed the newborn hearing screening(otoacoustic emission)and was diagnosed with mod-erate sensorineural hearing loss through audiology evaluation.Follow-up tests of blood calcium and parathyroid hor-mone levels were normal,and ultrasound examinations of the urinary system and parathyroid gland showed no ab-normalities.Patient 2 passed the newborn hearing screening but failed the 3-year-old physical examination(otoa-coustic emission)and was diagnosed with moderate sensorineural hearing loss.Follow-up tests of blood calcium and parathyroid hormone levels were normal,and the parathyroid gland ultrasound showed no abnormalities,but the re-nal ultrasound showed bilateral small renal calculi with normal morphology.Both patients were diagnosed with HDR syndrome through gene testing,and the 2 GAT A3 gene mutation sites(c.867dup,c.65_68dup)causing the disease were both reported for the first time.Conclusion The clinical phenotypes of HDR syndrome are highly variable.Children with suspected hearing loss accompanied by hypoparathyroidism or renal dysfunction should have gene tes-ting and other related examinations as soon as possible to avoid misdiagnosis.
8.CACNA1C rs58619945 genotype influences the cortical thickness of attention network among patients with Bipolar Ⅰ disorder
Xiaofei ZHANG ; Xiaoyu LIANG ; Xiaofang CHENG ; Jianshan CHEN ; Wenhao DENG ; Yizhi ZHANG ; Liqian CUI ; Xiongchao CHENG ; Xuan LI ; Chanjuan YANG ; Biyu YE ; Liping CAO
Chinese Journal of Medical Genetics 2024;41(9):1045-1052
Objective:To explore the impact of CACNA1C rs58619945 genotype on the cortical thickness of attentional networks in patients with Bipolar 1 disorder type (BD-Ⅰ). Methods:From August 2013 and August 2019, a total of 155 BD-Ⅰ patients were recruited from the outpatient and inpatient Departments of the Affiliated Brain Hospital of Guangzhou Medical University, along with 82 healthy controls (HC) from the community and university. Genotype for the CACNA1C rs58619945 locus was determined for all BD-I patients and HC subjects, followed by 3.0 T magnetic resonance imaging scans to measure the cortical thickness in the alert, orienting, and executive control subnetworks. General linear models (GLMs) were used to evaluate the impact of CACNA1C rs58619945 on the cortical thickness of attentional networks. Concurrently, attentional dimension functions were assessed using repeatable battery for the assessment of neuropsychological status (RBANS) and Cambridge neuropsychological test automated battery rapid visual information processing (CANTAB RVP) test. This study was approved by the Medical Ethics Committee of the Affiliated Brain Hospital of Guangzhou Medical University(Ethics No. 2023-056). Results:Compared with the HC group, the BD-Ⅰ patients had shown reduced thickness in bilateral prefrontal cortex, bilateral posterior cingulate cortex, and bilateral superior temporal cortex( P<0.05). A significant interaction between the CACNA1C genotype and the cortical thickness(HC vs.BD) of right prefrontal cortex, right posterior parietal cortex and right superior temporal cortex was noted( P<0.05). Partial correlation analysis has demonstrated a significant correlation between CANTAB RVP and RBANS attention indices and cortical thickness in the right prefrontal cortex, right posterior cingulate cortex( P<0.05), and right superior temporal cortex predominantly among carriers of the BD-Ⅰ G allele. Conclusion:The G allele of CACNA1C rs58619945 is associated with cortical thickness of the right prefrontal cortex, right posterior cingulate cortex, and right superior temporal cortex in BD-Ⅰ, which are part of the alerting and orienting network.
9.Hematopoietic aging: Cellular, molecular, and related mechanisms
Li YE ; Chuan TIAN ; Ye LI ; Hang PAN ; Jinxiu HU ; Liping SHU ; Xinghua PAN
Chinese Medical Journal 2024;137(11):1303-1312
Aging is accompanied by significant inhibition of hematopoietic and immune system function and disruption of bone marrow structure. Aging-related alterations in the inflammatory response, immunity, and stem cell niches are at the root of hematopoietic aging. Understanding the molecular mechanisms underlying hematopoietic and bone marrow aging can aid the clinical treatment of aging-related diseases. In particular, it is unknown how the niche reprograms hematopoietic stem cells (HSCs) in an age-dependent manner to maintain normal hematopoiesis in elderly individuals. Recently, specific inhibitors and blood exchange methods have been shown to reshape the hematopoietic niche and reverse hematopoietic aging. Here, we present the latest scientific discoveries related to hematopoietic aging and hematopoietic system rejuvenation, discuss the relationships between hematopoietic niche aging and HSC aging, and describe related studies on stem cell-mediated regulation of hematopoietic aging, aiming to provide new ideas for further study.
10.Effects of monocyte chemoattractant protein-1 on invasion and migration of lung cancer A549 and their mechanisms
Yuan WANG ; Zhijuan WANG ; Mingshu ZHANG ; Yihui WANG ; Qing ZHANG ; Liping YE
Journal of Jilin University(Medicine Edition) 2024;50(3):666-675
Objective:To discuss the effects of monocyte chemoattractant protein-1(MCP-1)on the migration and invasion of lung cancer A549 cells,and to clarify the mechanisms.Methods:Immunohistochemistry method was used to detect the expression of MCP-1 protein in 80 cases of non-small cell lung cancer(NSCLC)and adjacent normal lung tissues.The human lung cancer A549 cells were cultured in vitro.The MCP-1-small interfering RNA(siRNA)experiment was divided into blank group,negative control group(si-NC group),MCP-1-siRNA-1 group,and MCP-1-siRNA-2 group.The MCP-1 over-expression experiment was divided into control group,empty vector control group(OE-NC,transfected with MCP-1 over-expression empty vector),over-expression MCP-1 group(OE-MCP-1 group,transfected with MCP-1 over-expression plasmid),over-expression MCP-1+extracellular regulated protein kinase(ERK)pathway inhibitor PD98059 group(OE-MCP-1+PD98059 group,co-transfected with MCP-1 over-expression plasmid and PD98059),and PD98059 group(transfected with PD98059).The MCP-1 siRNA and plasmids were transfected into the lung cancer A549 cells;Western blotting method was used to verify the transfection efficiencies of the cells in various groups;the migration rate and the number of invasion cells in various groups were observed by wound healing assay and Transwell chamber assay,respectively;Western blotting method was also used to detect the expression levels of phosphorylated ERK(p-ERK),total ERK(t-ERK),and epithelial-mesenchymal transition(EMT)-related proteins in the A549 cells in various groups.Results:Compared with adjacent tissue,the positive expression rate of MCP-1 protein in NSCLC tissue was significantly increased(P<0.05),and the expression level of MCP-1 protein was related to TNM stage and lymph node metastasis(P<0.05).Compared with si-NC group,the expression level of MCP-1 protein in the cells in MCP-1-siRNA-1 and MCP-1-siRNA-2 groups was significantly decreased(P<0.01).Compared with control group and OE-NC group,the expression level of MCP-1 protein in the cells in OE-MCP-1 group was significantly increased(P<0.01).The wound healing assay results showed that compared with si-NC group,the migration rate of the cells in MCP-1-siRNA-1 and MCP-1-siRNA-2 groups were significantly decreased(P<0.01).Compared with OE-NC group,the migration rate of the cells in OE-MCP-1 group was significantly increased(P<0.01);compared with OE-MCP-1 group,the migration rate of the cells in OE-MCP-1+PD98059 group was significantly decreased(P<0.01).Compared with OE-MCP-1+PD98059 group,the migration rate of the cells in PD98059 group was significantly decreased(P<0.01).The Transwell chamber assay results showed that compared with si-NC group,the number of invasion cells in MCP-1-siRNA-1 and MCP-1-siRNA-2 groups was significantly decreased(P<0.01).Compared with OE-NC group,the number of invasion cells in OE-MCP-1 group was significantly increased(P<0.01);compared with OE-MCP-1 group,the number of invasion cells in OE-MCP-1+PD98059 group was significantly decreased(P<0.01);compared with OE-MCP-1+PD98059 group,the number of invasion cells in PD98059 group was significantly decreased(P<0.01).The Western blotting results showed that compared with si-NC group,the expression levels of p-ERK,Vimentin,and N-cadherin protein in the cells in MCP-1-siRNA-1 and MCP-1-siRNA-2 groups were significantly decreased(P<0.05 or P<0.01),and the expression level of E-cadherin proteins was significantly increased(P<0.01).Compared with OE-NC group,the expression levels of p-ERK,Vimentin,and N-cadherin proteins in the cells in OE-MCP-1 group were significantly increased(P<0.01),and the expression level of E-cadherin protein was significantly decreased(P<0.01).Compared with OE-MCP-1 group,the expression levels of p-ERK,Vimentin,and N-cadherins proteins in the OE-MCP-1+PD98059 group were significantly decreased(P<0.01),and the expression level of E-cadherin protein was significantly increased(P<0.05).Compared with OE-MCP-1+PD98059 group,the expression levels of p-ERK,Vimentin,and N-cadherin proteins in the cells in PD98059 group were significantly decreased(P<0.05 or P<0.01),and the expression level of E-cadherin protein was increased(P<0.01).Conclusion:MCP-1 protein can upregulate the expression of EMT-related proteins in the lung cancer A549 cells,and promote the migration and invasion of the lung cancer A549 cells;its mechanism may be related to the activation of the ERK signaling pathway.


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