1.Expression of miRNA-181 a in human lung adenocarcinoma cells and its effect on cell function
Duo ZHAO ; Zhu JIN ; Yaya SONG ; Baoan GAO
Chinese Journal of Pathophysiology 2016;32(8):1395-1402
AIM:To study the expression of microRNA (miRNA)-181a in different human lung adenocarcinoma cell lines, and to investigate the effect of miRNA-181a on cell function and its mechanism in human lung adenocarcinoma drug resistant cell A549/DDP.METHODS:Real-time PCR was used to detect the expression of miRNA-181a in BEAS-2B cells, A549 cells and A549/DDP cells.The A549/DDP cells were transfected with pGenesil-miRNA-181a eukaryotic ex-pression plasmid.At the same time, the untransfection group and negative transfection group were also set up .The expres-sion of miRNA-181a, cell viability, cell growth inhibition and apoptosis rate during cis-diamminedichloroplatinum ( DDP) treatment, cell cycle, cell invasion, the protein expression of miRNA-181a target genes bcl-2 and p53 in the A549/DDP cells were determined by real-time fluorescence quantitative PCR , MTT assay, flow cytometry, Transwell method and West-ern blot, respectivly.RESULTS:The expression of miRNA-181a in A549 cells and A549/DDP cells was significantly lower than that in BEAS-2B cells, and the lowest expression level was observed in A 549/DDP cells (P<0.05).The expression of miRNA-181a in A549/DDP cells was significantly increased after transfection with pGenesil-miRNA-181a (P<0.05).The cell viability, cell cycle and invasion ability of the A549/DDP cells were inhibited after miRNA-181a transfection (P <0.05).The cell growth inhibition rate and apoptotic rate of the A 549/DDP cells were increased (P<0.05).The expression of Bcl-2 was reduced, but the expression of P53 was increased after transfection with miRNA-181a in A549/DDP cells (P<0.05).CONCLUSION: miRNA-181a may be correlated with the development of human lung adenocarcinoma .miRNA-181a can serve as a new target for treatment of lung cancer .
2.Effect of dose rate of X-ray on clonogenic formation in human lung cancer cell line A549
Shujun SONG ; Shaoyan SI ; Yaya QIN ; Xiaoyong ZUO ; Gaixian SHAN ; Ye REN ; Zongye WANG
Cancer Research and Clinic 2017;29(2):83-85
Objective To explore the effects of different dose rates of X-ray under the same dose on cell clonogenic formation in non-small-cell lung cancer cell line A549 in order to provide experimental basis for clinical radiotherapy plan. Methods The A549 cells were cultured at low density and irradiated with X-rays at dose of 4 Gy and selected dose rates of 1, 2, 4 and 6 Gy/min, respectively, from a linear accelerator. The 8th day after irradiation, the cells were fixed and stained with Giemsa solution, and colonies containing at least 50 cells were counted. The plating efficiency and surviving fraction were calculated. Results The clonogenic number in non-irradiated cells was 88.6±4.6. The numbers were significantly reduced in irradiated cells at dose rate 1, 2, 4 and 6 Gy/min (12.3±3.4, 9.0±0.8, 5.6±1.0, 11.5±1.7, respectively) than that in non-irradiated control cells (F=678.799, P<0.05). The plating efficiencies were decreased in irradiated cells, especially in 4 Gy/min irradiated cells, which was lower than that in any of the other three dose rate groups (P< 0.05). Conclusions Though at same radiation dose, cancer cells have different clonogenic formation efficiency when irradiation with X-ray at different dose rates. Thus, treatment with optimal dose rate may improve the radiotherapy efficacy.
3.Establishment of human acute myeloblastic Leukemia M2 type nude mouse model
Yuan LIU ; Ruihong DONG ; Yuhua LI ; Yaya WANG ; Jing WU ; Rui HUANG ; Lan DENG ; Zhaoyang SONG ; Zhigang LU ; Haiyan HU
Chinese Journal of Tissue Engineering Research 2010;14(1):95-98
BACKGROUND: It is easy to established human solid tumor nude mouse model, but for leukemia which is difficult. We inhibited immune system further by radioactive ray or CTX, to decrease cost and increase the stability.OBJECTIVE: To establish a human acute myeloblastic leukemia M2 Kasumi-1 models containing AML/ETO positive genes in BALB/c nude mouse. METHODS: Nude mice were randomly divided into three groups: CTX group was injected CTX 2 mg/day in abdominal cavity for two days, and injected 8×10~5/mouse Kasumi-1 cells in caudal vein next day; irradiation group was exposed to total body irradiation, and injected 8×10~5/mouse Kasumi-1 cells in caudal vein that day; untreated group was inoculated with 8×10~5/mouse Kasumi-1 cells by caudal vein injection. Three additional mice were considered as the normal control group. The blood smearing and bone morrow slides were detected, immunity type of BMC was detected using flow cytometry, loading of leukemic cellular tumor was detected using RT-PCR, and positive ratio of AML/ETO fusion gene was detected using FISH method. RESULTS AND CONCLUSION: After inoculated into untreated nude mice by caudal vein injection for 14 days, the ratio of leukemia cell in blood smearing was 3.5%, and over 40% in bone marrow slides, which was equal to the results of FISH and FCM. The increasing of tumor loading was time-dependent. For irradiation group and CTX treated group, the tumor loading was higher that untreated group, and the cells also survived more than 60 days. AML/ETO band was observed by RT-PCR in all experimental groups, for normal mice it was negative. The results indicated that the systemic disseminated leukemia model was established successfully by caudal vein injection 8×10~5/mouse Kasumi-1 cells in the three experimental groups.
4.Mesenchymal Stem Cell-Derived Exosomes are Effective for Radiation Enteritis and Essential for the Proliferation and Differentiation of Lgr5+ Intestinal Epithelial Stem Cells by Regulating Mir-195/Akt/b-Catenin Pathway
Leilei YANG ; Chengfeng FANG ; Caifang SONG ; Yaya ZHANG ; Ruili ZHANG ; Shenkang ZHOU
Tissue Engineering and Regenerative Medicine 2023;20(5):739-751
BACKGROUND:
Radiation enteritis (RE) is a common complication of abdominal or pelvic radiotherapy, which when severe, could be life-threatening. Currently, there are no effective treatments. Studies have shown that mesenchymal stem cells (MSCs)-derived exosomes (MSC-exos) exhibit promising therapeutic effects in inflammatory diseases. However, the specific role of MSC-exos in RE and the regulatory mechanisms remain elusive.
METHODS:
In vivo assay was carried out by injecting MSC-exos into the total abdominal irradiation (TAI)-induced RE mouse model. For in vitro assay, Lgr5-positive intestinal epithelial stem cells (Lgr5+ IESC) were extracted from mice, followed by irradiation along with MSC-exos treatment. HE staining was performed to measure histopathological changes. mRNA expression of inflammatory factors TNF-a and IL-6 and stem cell markers LGR5, and OCT4 were quantified by RT-qPCR. EdU and TUNEL staining was performed to estimate cell proliferation and apoptosis. MiR-195 expression in TAI mice and radiation-induced Lgr5+ IESC was tested.
RESULTS:
We found that the injection of MSC-exos inhibited inflammatory reaction, increased stem cell marker expression, and maintained intestinal epithelial integrity in TAI mice. Furthermore, MSC-exos treatment increased the proliferation and simultaneously suppressed apoptosis in radiation-stimulated Lgr5+ IESC. MiR-195 expression increased by radiation exposure was decreased by MSC-exos therapy. MiR-195 overexpression facilitated the progress of RE by counteracting the effect of MSC-exos. Mechanistically, the Akt and Wnt/b-catenin pathways inhibited by MSC-exos were activated by miR-195 upregulation.
CONCLUSION
MSC-Exos are effective in treating RE and are essential for the proliferation and differentiation of Lgr5+ IESCs. Moreover, MSC-exos mediates its function by regulating miR-195 Akt b-catenin pathways.
5.Research status and prospect of tissue engineering technology in treatment of atrophic rhinitis.
Shuting LEI ; Juanjuan HU ; Yingqi TANG ; Weigang GAN ; Yuting SONG ; Yanlin JIANG ; Honghui ZHANG ; Yaya GAO ; Hui YANG ; Huiqi XIE
Chinese Journal of Reparative and Reconstructive Surgery 2023;37(6):727-731
OBJECTIVE:
To review the research progress of the feasibility of a new treatment method for atrophic rhinitis (ATR) based on tissue engineering technology (seed cells, scaffold materials, and growth factors), and provide new ideas for the treatment of ATR.
METHODS:
The literature related to ATR was extensively reviewed. Focusing on the three aspects of seed cells, scaffold materials, and growth factors, the recent research progress of ATR treatment was reviewed, and the future directions of tissue engineering technology to treat ATR were proposed.
RESULTS:
The pathogenesis and etiology of ATR are still unclear, and the effectiveness of the current treatments are still unsatisfactory. The construction of a cell-scaffold complex with sustained and controlled release of exogenous cytokines is expected to reverse the pathological changes of ATR, promoting the regeneration of normal nasal mucosa and reconstructing the atrophic turbinate. In recent years, the research progress of exosomes, three-dimensional printing, and organoids will promote the development of tissue engineering technology for ATR.
CONCLUSION
Tissue engineering technology can provide a new treatment method for ATR.
Humans
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Tissue Engineering/methods*
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Tissue Scaffolds
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Rhinitis, Atrophic
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Printing, Three-Dimensional
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Cytokines
6.PRMT6 promotes tumorigenicity and cisplatin response of lung cancer through triggering 6PGD/ENO1 mediated cell metabolism.
Mingming SUN ; Leilei LI ; Yujia NIU ; Yingzhi WANG ; Qi YAN ; Fei XIE ; Yaya QIAO ; Jiaqi SONG ; Huanran SUN ; Zhen LI ; Sizhen LAI ; Hongkai CHANG ; Han ZHANG ; Jiyan WANG ; Chenxin YANG ; Huifang ZHAO ; Junzhen TAN ; Yanping LI ; Shuangping LIU ; Bin LU ; Min LIU ; Guangyao KONG ; Yujun ZHAO ; Chunze ZHANG ; Shu-Hai LIN ; Cheng LUO ; Shuai ZHANG ; Changliang SHAN
Acta Pharmaceutica Sinica B 2023;13(1):157-173
Metabolic reprogramming is a hallmark of cancer, including lung cancer. However, the exact underlying mechanism and therapeutic potential are largely unknown. Here we report that protein arginine methyltransferase 6 (PRMT6) is highly expressed in lung cancer and is required for cell metabolism, tumorigenicity, and cisplatin response of lung cancer. PRMT6 regulated the oxidative pentose phosphate pathway (PPP) flux and glycolysis pathway in human lung cancer by increasing the activity of 6-phospho-gluconate dehydrogenase (6PGD) and α-enolase (ENO1). Furthermore, PRMT6 methylated R324 of 6PGD to enhancing its activity; while methylation at R9 and R372 of ENO1 promotes formation of active ENO1 dimers and 2-phosphoglycerate (2-PG) binding to ENO1, respectively. Lastly, targeting PRMT6 blocked the oxidative PPP flux, glycolysis pathway, and tumor growth, as well as enhanced the anti-tumor effects of cisplatin in lung cancer. Together, this study demonstrates that PRMT6 acts as a post-translational modification (PTM) regulator of glucose metabolism, which leads to the pathogenesis of lung cancer. It was proven that the PRMT6-6PGD/ENO1 regulatory axis is an important determinant of carcinogenesis and may become a promising cancer therapeutic strategy.