1.Effect of silencing cyclin D1 gene on chemotherapeutic drug sensitivity in K562 cells in vitro
Yiyuan WAN ; Yangqing LIU ; Xiaofei CHEN ; Wenqi JIANG
Cancer Research and Clinic 2008;20(3):148-151
Objective Cyclin D1 gene plays a significant role in regulating cell cycle progression.It is reported that over-expression of cyclin D1 gene is intimately associated with origination,development and prognosis of tumor and is associated with tumor cells resistance to chemotherapy drug.Suppression of cyclin D1 protein expression leads to cellular chemosensitization.This study was to determine whether this effect also existed in chronic leukemia cell line K562 by inhibiting the expression of cyclin D1 protein through RNA interference.Methods Plasmid vectors expressing small hairpin RNA (shRNA) targeting at cyclin D1 gene were constructed and transfected into K562 cells by chitosan.Cyclin D1 protein was examined using Western Blotting analysis.The cell cycle and apoptosis were determined by flow cytometry.Cellular chemosensitization was evaluated by MTY assay.Results Expression of cyclin D1 protein was markedly down-regulated after transfection with pshRNA-419 and pshRNA-575 at 48 h.Down-regulation of cyclin D1 protein could affect the redistribution of cell cycle,induce apoptosis of K562 cells,decrease 50%inhibitoryconcentration (IC50) of adriamycin and enhance cellular chemosensitization.But there had no above biological effects observed after transfection with blank vector and control vector of m-pshRNA-790 at 48 h.ConclusionK562 cells could be chemosensitized by the down-regulation of cyclin D1 expression through RNA interference.
2.METTL3 promotes proliferation,migration,and secretion of inflammatory factors by mRNA m6A methylation in rheumatoid arthritis synovial fibroblasts
Juan LI ; Yangqing JIANG ; Ruiming SHEN ; Guoquan LI ; Min WANG ; Fenghuang XU
Acta Universitatis Medicinalis Anhui 2024;59(4):619-626
Objective To investigate the effect and mechanism of methyltransferase-like 3(METTL3)on the pro-liferation,migration,and secretion of inflammatory factors by synovial fibroblasts from rheumatoid arthritis(RA).Methods The expression of METTL3 in synovial tissue(SF)from 25 patients with rheumatoid arthritis and 25 pa-tients with osteoarthritis was detected by RT-qPCR and immunohistochemistry,respectively.The concentration of RNA m6A was detected by ELISA.RA synovial fibroblasts were isolated and cultured,and divided into NC(nor-mal control)group,hi-METTL3(overexpression of METTL3)group,si-METTL3(knock-down METTL3)group,and STM2457(METTL3 specific inhibitor)intervention group.Cell proliferation was detected by CCK-8 method.Apoptosis was detected by flow cytometry.And the concentrations of interleukin-6(IL-6),interleukin-17A(IL-17A),receptor activator of nuclear factor-kappa B ligand(RANKL),and osteoprotegerin(OPG)in the superna-tant of cell culture were detected by ELISA.Results Compared with synovial tissue of osteoarthritis,the expres-sion of mRNA m6A and METTL3 in synovial tissue of RA significantly increased(P<0.05).After overexpression of METTL3,the expression of m6A in synovial fibroblasts increased.The proliferation and migration abilities of SF in hi-METTL3 group were significantly improved,and their apoptosis did not change significantly.The secretion of cytokines IL-6 and RANKL of SF in hi-METTL3 group significantly increased,while the OPG significantly de-creased(P<0.05).After interfering with METTL3 expression,the expression of m6A in synovial fibroblasts de-creased.Cell proliferation and migration of SF in siMETTL3 group significantly decreased.The secretion of cyto-kines IL-6 and RANKL significantly decreased,and OPG significantly increased(P<0.05).After intervention with METTL3 inhibitor STM2457,the proliferation and migration of synovial fibroblasts were significantly reduced,and the secretion of cytokines IL-6 and RANKL significantly reduced,and OPG significantly increased(P<0.05).There was no significant difference in the expression of IL-17A among each group.Conclusion METTL3 may promote the proliferation and migration of RA synovial fibroblasts,enhance the expression of IL-6 and RANKL,and inhibit the expression of OPG through RNA m6A methylation modification.
3.Effects of feedback method education model in patients with recurrent aphthous ulcer
Yangqing ZHANG ; Huiming YE ; Qiyue CHEN ; Xiaoying JIANG ; Xiaoyun ZHU
Chinese Journal of Modern Nursing 2021;27(21):2878-2882
Objective:To explore the effect of the feedback method education model in patients with recurrent aphthous ulcer (RAU) , so as to provide a new education model for clinical health education.Methods:From May 2019 to May 2020, convenience sampling was used to select 76 RAU patients in the Stomatology Hospital of Southern Medical University. According to the method of random number table, the patients were divided into the experimental group and the control group, with 38 cases in each group. The experimental group implemented feedback method education model, and the control group carried out routine health education. The scores of Morisky Medication Adherence Scale (MMAS-8) , number of ulcers, healing time, pain degree, ulcer diameter and the Oral Health Impact Profile (OHIP-14) scores of patients were compared between the two groups before and after the intervention.Results:The score of MMAS-8 in the experimental group was higher than that in the control group after the intervention, and the difference was statistically significant ( P<0.05) . After the intervention, the number of ulcers in the experimental group was less than that in the control group, and the ulcer diameter was smaller than that in the control group, and the healing time was shorter than that in the control group, and the pain degree was lower than that in the control group, and the differences were statistically significant ( P<0.05) . After the intervention, the total score and dimension scores of OHIP-14 in the experimental group were lower than those in the control group, and the differences were also statistically significant ( P<0.05) . Conclusions:The feedback method education model can improve the adherence of RAU patients, the clinical indicators and the quality of life related to oral health of patients, which is worthy of clinical promotion and application.