1.Effect and mechanism of LncRNA PICSAR knockdown on proliferation,migration and apoptosis of ovarian cancer cells
Xuejun TANG ; Xiaowei DOU ; Xiaoyan YING
Acta Universitatis Medicinalis Anhui 2024;59(9):1509-1516
Objective To investigate the expression of long non-coding RNA(LncRNA)PICSAR in ovarian canc-er,and explore the effects of LncRNA PICSAR on the proliferation,migration,invasion and apoptosis of ovarian cancer cells as well as its possible mechanism of action.Methods The expression levels of LncRNA PICSAR in o-varian cancer tissue and cell line A2780,OVCAR-3,HO-8910 and normal ovarian tissue and cell line IOSE386 were detected by fluorescence quantitative PCR.Ovarian cancer cell lines with the highest expression of LncRNA PICSAR were divided into control group and experimental group,and transfected with negative control small inter-fering RNA(siRNA-NC)or PICSAR knockout small interfering RNA(siRNA-PICSAR)by liposome transfection technique,respectively.The effects of LncRNA PICSAR knockdown on the invasion,migration,proliferation and apoptosis of ovarian cancer cells were analyzed by cell counting assay(CCK-8),clonogenic assay,scratch assay,transwell assay and flow cytometry and so on.The expression levels of autophagy related proteins and apoptosis-re-lated proteins in each group were determined by Western blot.Ovarian cancer cells were treated with rapamycin and hydroxychloroquine as autophagy activator and inhibitor,and Western blot assay was used to detect apoptosis.Results The expression level of LncRNA PICSAR in ovarian cancer tissues was higher than that in normal ovarian tissues.Compared with IOSE386 cell line,LncRNA PICSAR expression levels in ovarian cancer cell lines HO-8910,OVCAR-3 and A2780 increased.Compared with the si-NC group,the proliferation,invasion and migration ability of ovarian cancer cells in si-PICSAR group decreased,and the apoptosis rate increased.The autophagy level of ovarian cancer cells in si-PICSAR group was lower than that in si-NC group.After transfection with siRNA-PIC-SAR,rapamycin activated autophagy to reduce apoptosis,while hydroxychloroquine inhibited autophagy to promote apoptosis.Conclusion LncRNA PICSAR is highly expressed in ovarian cancer tissues and cell lines,and the ma-lignant biological behavior of ovarian cancer cells can be inhibited by knockout of LncRNA PICSAR.The knock-down of LncRNA PICSAR may promote the apoptosis of ovarian cancer cells by regulating autophagy.
2.Establishment of primary breast cancer cell line as new model for drug screening and basic research
Xian HAO ; Jianjun HUANG ; Wenxiu YANG ; Jinting LIU ; Junhong ZHANG ; Yubei LUO ; Qing LI ; Dahong WANG ; Yuwei GAO ; Fuyun TAN ; Li BO ; Yu ZHENG ; Rong WANG ; Jianglong FENG ; Jing LI ; Chunhua ZHAO ; Xiaowei DOU
China Oncology 2024;34(6):561-570
Background and purpose:In 2016 the National Cancer Institute(NCI)decided stopping to use NCI-60 cell lines for drug screening,suggesting that tumor cell lines were losing their value as a tool for drug discovery and basic research.The reason for NCI-60 cells'retirement'was that the preclinical studies based on traditional cellular and animal models did not obtain the corresponding expected efficacy in clinical trials.Since the major cancer behaviors,such as proliferation and metastasis,are fundamentally altered with long-term culture,the tumor cell lines are not representative of the characteristics of cancer in patients.Currently,scientists hope to create a new cancer model that are derived from fresh patient samples and tagged with details about their clinical past.Our purpose was to create patient-derived breast cancer primary cell lines as new cancer model for drug screening and basic research.Methods:Breast cancer tissues were collected in the Department of Breast Surgery,Affiliated Hospital of Guizhou Medical University.The collection of tumor tissue samples was approved by the Ethics Committee of the Affiliated Hospital of Guizhou Medical University(approval number:2022 ethics No.313),and the collection and use of tumor tissues complied with the Declaration of Helsinki.The primary breast cancer cell lines were isolated from the patient's breast cancer tissues and cultured in BCMI medium.After the cells proliferated,the media were replaced with DEME medium.Cell line STR genotyping was done to determine cell-specific genetic markers and identification.Clone formation assay and transplantation assay were done to analyze the ability of breast cancer primary cell lines to form tumors.Results:We created 6 primary breast cancer cell lines.The 6 primary breast cancer cell lines from the patients were tagged with the definitively clinicopathological features,clinical diagnosis,therapeutic regimens,clinical effectiveness and prognostic outcomes.The STR genotyping assays identified the genetic markers and determined the identities of the 6 primary breast cancer cell lines.Clone formation assays and transplantation assay showed that the proliferative capacities of the patient-derived primary breast cancer cell lines were significantly greater compared with the conventional breast cancer cell lines.Conclusion:We created a panel of 6 patient-derived primary breast cancer cell lines as new cancer model for drug screening and basic research in breast cancer.
3.Research Progress in Chinese Materia Medica Regulation of Glucose Metabolism Reprogramming Intervention in Gastric Precancerous Lesions Based on Hypoxia Microenvironment
Xiaolong WANG ; Ruiping SONG ; Pengcheng DOU ; Zhuangzhuang FENG ; Xiaowei SUN ; Dongxu LEI ; Jing YANG ; Qingshan NAN ; Jin SHU
Chinese Journal of Information on Traditional Chinese Medicine 2024;31(9):179-184
Precancerous lesions of gastric cancer is a key stage in the development of gastric cancer.The reprogramming of glucose metabolism is a prominent feature of precancerous lesions of gastric cancer.Hypoxic microenvironment and hypoxia-inducible factors are important factors influencing the occurrence of glucose metabolic reprogramming.This article summarized the relationship between hypoxic microenvironment and the reprogramming of glucose metabolism in precancerous lesions of gastric cancer,and concluded the relevant research on TCM compounds and effective components to improve hypoxic microenvironment and further regulate glycolysis for the treatment of this disease.It was concluded that the mechanism may be the inhibition of angiogenesis,regulation of signaling pathways and key proteins of glycolysis,expression of multiple enzymes,reduction of lactate secretion,inhibition of cell malignant proliferation and invasion.It explored the mechanism of Chinese materia medica in improving hypoxic microenvironment and regulating glycolysis,so as to provide reference for the prevention and treatment of precancerous lesions of gastric cancer.
4.Evaluation on the application of light scattering method to detect hemoglobin in lipoblood samples
Min DOU ; Yanhui LIN ; Xiaowei GUO ; Pengju CAO
Chinese Journal of Laboratory Medicine 2021;44(12):1182-1189
Objective:To evaluate the ability of light dispersive colorimetry to detect hemoglobin (Hb) in lipid blood samples and its feasibility as an alternative to plasmapheresis commonly used in the laboratory.Methods:Routine blood samples of 276 inpatients in Fujian Provincial Hospital from July 2020 to July 2021 were collected. Routine blood samples of 276 inpatients were collected. There were 169 males and 107 females, aged from 16 to 97 years. 183 non-lipid blood samples and 93 lipid blood samples were collected.(1) One case each of low, medium and high Hb value in non-lipid blood and lipid blood samples were collected, and the precision and the linearity of light scattering method was detected.(2)Non-lipid blood samples divided into Hb low-value group, median-value group and high-value group, which were measured by light scattering method and colorimetric method to compare Hb values. (3)Non-lipid blood samples were divided into Hb low-value group, median-value group and high-value group. Plasma exchange was carried out with different concentrations of fat emulsion. The bias and linearity of Hbc2 and Hbc1, Hbo2 and Hbc1 were analyzed by MedCalc19.1 software. The Hbc2 and Hbc1 bias ( CV%) and Hbo2 and Hbc1 bias ( CV%) were calculated. T test was used to analyze the influence of different concentrations of triglyceride on Hb bias.(4)Blood samples were divided into Hb low-value group, median-group and high-value group. The Hb of light scattering method was compared with the colorimetric method after plasma exchange. Results:(1)The intrabatch precision of light scattering method for non-lipid blood and lipid blood specimens was within the allowable range ( CV<1.5%), and the good linearity ( R2=1.000).(2)The bias of Hb measured by light scattering method and colorimetric method in the three groups was below 3.5%(-0.58±2.34,0.16±1.52,1.15±1.56), within the allowable total error range. The two methods have the equivalence and good linear relationship ( r=0.999).(3)The concentrations of Hbo2 in the low (except 4.1 mmol/L), medium and high Hbo2 groups were equivalent to those in the non-lipid blood colorimetry (Hbc1), and the two methods were well correlated. The results of light scattering method have nothing to do with the concentration of lipid blood.(4)There was no significant difference of the Hb between the light scattering method and plasma exchange method in three groups ( P>0.05), Both of them have equivalence and good correlation ( R2=0.968,0.948,0.870). Conclusion:Light scattering method can effectively reduce the effect of lipid blood on hemoglobin determination, and can replace the traditional plasma exchange method, which has high clinical application value.
5.Effect of tantalum coating on adhesion, proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells
Xiaowei WEI ; Dewei ZHAO ; Ge LIU ; Xiaojie DOU ; Zhenhua ZHAO ; Weimin FU ; Tianzhu LIU
Chinese Journal of Orthopaedic Trauma 2020;22(5):438-444
Objective:To investigate the effects of tantalum coating on adhesion, proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) in vitro. Methods:In this study, BMSCs were extracted from 6 6-week-old rats and cultured in vitro to the third generation. Tantalum coating was manufactured on Ti6Al4V by chemical vapor deposition. The cells were identified by flow cytometry before they were induced with different mediums for osteogenesis, chondrogenesis and adipogenesis. The adhesion, proliferation and osteogenic differentiation of BMSCs were detected with fluorescence staining, Cell Counting Kit-8 (CCK8) assay and Q-PCR, respectively. Recorded and compared were the adhesion rate, proliferation rate, and expression of osterix (OSX), Runt-related transcription factor 2 (RUNX2), osteonectin (OSN) and osteopontin (OPN) of BMSCs on the surface of titanium alloy round plates (the Ti6Al4V group) and of tantalum coating round plates (the Ta group). Results:The flow cytometry revealed CD44 (94.55%), CD90 (95.01%) and CD34 (0.06%). Alkaline phosphatase (ALP) staining was positive after osteogenic induction for 14 days; Alizarin red staining showed calcified nodules after osteogenic induction for 21 days; oil red O staining was positive after adipogenic induction for 21 days; alcian blue staining found chondrogenic ability after chondrogenic induction for 21 days. Laser confocal microscopy showed that the BMSCs grew in patches aggregated and closely linked on the surface of titanium alloy round plates (in the Ti6Al4V group) and of tantalum coating round plates (in the Ta group). More BMSCs adhered on the tantalum coating plates than on the titanium alloy plates and exhibited better ductility. The proliferation rates of BMSCs on tantalum coating were significantly faster than those on titanium alloy after 1, 3, 5 and 7 days of co-culture in vitro ( P<0.05).Q-PCR showed that tantalum coating promoted the expression of OSN and OPN after 7 days of culture significantly higher than titanium alloy did ( P<0.05).After 21 days of co-culture in vitro, tantalum coating enhanced the expression of OSX, RUNX2, OSN and OPN significantly higher than titanium alloy did ( P<0.05). Conclusion:Compared to titanium alloy which is used for conventional orthopedic implants, tantalum coating can observably promote adhesion, proliferation and osteogenic differentiation of BMSCs.
6.The clinical characteristics and angiographic findings of cardiogenic shock following acute myocardial infarction in elderly patients
Yan CHEN ; Mingdong GAO ; Xiaowei LI ; Haiwang ZHAO ; Nan ZHANG ; Jing DOU ; Yin LIU
Chinese Journal of Geriatrics 2016;35(9):939-943
Objective To investigate the clinical characteristics and angiographic findings of cardiogenic shock(CS)following acute myocardial infarction(AMI) in elderly patients.Methods Between January 2015 and April 2016,we carried out a retrospective observational analysis of consecutive elderly patients in Tianjin Chest Hospital,who suffered CS-complicating AMI.Emergency angiography and percutaneous coronary intervention(PCI) were performed after admission.All selected patients were divided into CS and non-CS groups according to whether CS occurred.Electrocardiograph (ECG),cardiac enzyme testing,and ultrasound cardiography were performed after admission to monitor the occurrence of CS.Results The incidence of CS-complicating AMI was 8.33% (34/408) in elderly patients.Among all CS patients enrolled,the aged patients accounted for 91.89 % (3 4/3 7).In-hospital mortality rate was 2 9.41 % (10/3 4).There were significant differences between two groups in WBC,H s-CRP,blood glucose,CR and ALT (t =2.403,4.596,6.778,6.109,each P<0.05).The NT-Pro BNP level,the time of FMC,the frequency of left main and multivessel disease were higher in the CS group than in the non-CS group (each P < 0.05).Conclusions Elderly patients are bearing high risk of CS following AMI.Prolonged FMC time and the presence of left main and/or multivessel lesion are independent risk factors for the development of CS.The optimal revascularisation strategy can improve the clinical outcome of patients with CS.
7.Creation and effectiveness of mammary stem cell medium
Junhong ZHANG ; Jing YANG ; Xiaowei DOU ; Chunhua WANG ; Qing LI ; Chunhua ZHAO
Chinese Journal of Tissue Engineering Research 2014;(10):1585-1590
BACKGROUND:Now in mammary stem cellresearch, no proper mammary stem cellmedium is provided to culture mammary stem cells. OBJECTIVE:To create a mammary stem cellmedium and validate its application by isolating Sca-1+mammary stem cells. METHODS:We first used BM medium to culture mammary organoids, and after 6 days, the expression of Sca-1 and vimentin was detected in fibroblasts by immunofluoresence method. Then, we established MaECM medium which arrested fibroblasts growth. After 6 days culture of mammary organoids by MaECM medium, Sca-1+and Sca-1-cellpopulations were sorted out by magnetic sorting and the purity was analyzed by flow cytometry. Sorted 1×104 Sca-1+or Sca-1-cells were transplanted into the bilateral mammary fat pads of four mice, and after 6-8 weeks, the fat pads were harvested for whole-mount immunohistochemical analysis and hematoxylin-eosin staining. RESULTS AND CONCLUSION:After 6 days culture of mammary organoids under BM medium, smal-sized colonies were generated around lots of fibroblasts. Immunofluoresence staining detected strong expression of vimentin and Sca-1 in fibroblasts, indicating that the BM medium is not suitable to isolate Sca-1+mammary stem cell. The MaECM medium promoted the proliferation of mammary epithelial cells whereas arrested fibroblasts growth. After 6 days culture of mammary organoids under MaECM medium and magnetic sorting, the flow cytometry showed that the purity of Sca-1+cellreached 92%and 5%in the Sca-1+and Sca-1-population, respectively. The results from transplantation test showed that six mammary outgrowths were regenerated out of eight injected fat pads in the Sca-1+cells transplantation, but in the Sca-1-transplantation population, one mouse died and the other transplants failed to produce outgrowths. We developed the MaECM medium which promoted the proliferation of mammary epithelial cells whereas arrested Sca-1+fibroblasts growth. Using the medium, we confirmed that Sca-1+mammary cells have capacity of isolating mammary stem cells.
8.Advance in Dynamic Neuromuscular Stabilization Training (review)
Jing PENG ; Xiaowei WANG ; Dongmei SUN ; Suhui MA ; Na DOU
Chinese Journal of Rehabilitation Theory and Practice 2014;(7):629-633
This article mainly discussed the concept and application of dynamic neuromuscular stabilization in order to make up for the defect that some of the traditional training only focus on large muscle groups, and ignore the deep trunk muscles, especially the small deep muscles.
9.Molecular targeted therapy for breast cancer patients based on Notch signal transduction pathway
Yongqu ZHANG ; Xiaowei DOU ; Guojun ZHANG
Journal of International Oncology 2013;40(9):678-680
Studies show that the Notch signal transduction pathway involves in the regulation and control of cell proliferation,differentiation and apoptosis.Aberrant Notch signaling transduction pathway can induce breast cancer in transgenic mice.High expressions of either Notch receptors or their ligands have been linked to poor prognosis in patients with breast cancer.Inhibition of Notch signal transduction pathway may be beneficial for breast cancer treatment.
10.The curative effect of nasal flush combined budesonide suspension liquid atomizing inhaled treatment in infant allergic rhinitis
Xiaowei SUN ; Yufang GAO ; Kebo GONG ; Fenfen DOU ; Zihe ZHANG ; Jianji ZHANG
Chinese Journal of Primary Medicine and Pharmacy 2011;18(3):313-314
Objective To observe the nasal flush combined budesonide suspension liquid atomizing inhaled treatment of allergic rhinitis in infants,explore the allergic rhinitis treatments in infants. Methods 137 cases diagnosed as allergic rhinitis were collected and randomly divided into 70 cases as the treatment group and 67 patients as the control group. The patients in control group were washed the nasal cavity with 2.8% warm sodium chloride solution using 50ml and 0.5% metronidazole injection 30ml by turn. At the first week, 1 time/d, then one time every other day,while according to age,body mass the patients were given to loratadine, 1 time/d. Treatment group were used budesonide nasal inhalation of 1 ml at the base of treatment of the control group. Before and after the treatment , nasal congestion,sneezing, flow clear nose, sleep snoring and sleep quality score index were observed and compared.Results 70%of the treatment group the nasal congestion,sneezing,flow clear in 3 times after treatment with ease.The children sleep quality improved and the snoring fewer over night,only 56.7% of the control group of these symptoms improved. After the treatment the efficiency evaluation of treatment group and control group respectively was 95.1% and 77.7% ,there was statistically significant difference( x2 =9.83 ,P <0. 01 ). 137 cases of patients without a side effects. Conclusion curative effect of nasal flush combined budesonide suspension liquid nasal spray inhaled treatment of allergic rhinitis was distinct,infant effect-acting quickly,without side effects,easy to use.


Result Analysis
Print
Save
E-mail