咪达唑仑调节YAP/TAZ信号通路对骨肉瘤细胞增殖凋亡和侵袭的影响

doi:10.3969/j.issn.1006-6233.2024.01.01

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摘要 目的: 探究咪达唑仑(MDZ)调节Yes相关蛋白(YAP)/PDZ结合基序转录共激活因子(TAZ)信号通路对骨肉瘤(OS)细胞增殖、凋亡和侵袭的影响。方法: qRT-PCR、Western blot检测人正常成骨细胞系、OS细胞系YAP、TAZ mRNA和蛋白的表达,筛选最佳干预细胞系;以不同浓度的MDZ(0、12.5、25、50、100、200μmoL/L)干预OS细胞,MTT法检测细胞增殖活性,筛选最佳干预浓度;将OS细胞随机分成control组、MDZ组、pcDNA3.1组、pcDNA3.1-YAP/TAZ组,qRT-PCR法检测转染效率;EdU染色、流式细胞仪和Transwell小室分别检测细胞增殖、凋亡和侵袭;Western blot方法检测增殖细胞核抗原(PCNA)、Bcl-2相关X蛋白(Bax)、N-钙黏蛋白(N-cadherin)、E-钙黏蛋白(E-cadherin)及YAP、TAZ蛋白表达;构建OS裸鼠模型,分为对照组和MDZ组,免疫组化法检测移植瘤组织Ki-67、YAP、TAZ蛋白表达,TUNEL染色检测凋亡。结果: OS细胞系中YAP、TAZ mRNA及蛋白表达水平显著升高(P<0.05);MDZ显著降低PCNA、N-cadherin、YAP、TAZ表达,抑制MG63、U20S细胞增殖和侵袭,升高Bax、E-cadherin表达,促进细胞凋亡。过表达YAP/TAZ可逆转MDZ对MG63、U20S细胞增殖和侵袭的抑制作用,对MG63、U20S细胞凋亡的促进作用(P<0.05);体内实验表明,MDZ显著降低移植瘤质量、体积及Ki-67、YAP、TAZ表达水平,促进移植瘤细胞凋亡(P<0.05)。结论: MDZ可能通过抑制YAP/TAZ信号通路,抑制OS细胞增殖和侵袭,促进凋亡。

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	奥婷婷
	涂梦佳
	张熙

关键词 ：咪达唑仑, YAP/TAZ, 骨肉瘤, 增殖, 凋亡, 侵袭

Abstract：Objective: To investigate the effects of midazolam (MDZ) on the proliferation, apoptosis and invasion of osteosarcoma (OS) cells by regulating the Yes associated protein (YAP)/PDZ transcriptional activator of transcription (TAZ) signaling pathway. Methods: qRT-PCR and western blot were applied to detect the expression of YAP, TAZ mRNA, and proteins in normal osteoblast and OS cell lines, and to screen the optimal intervention cell line. OS cells were intervened with different concentrations of MDZ (0, 12.5, 25, 50, 100, 200 μmoL/L), MTT method was applied to detect cell proliferation activity and screen the optimal intervention concentration; OS cells were randomly divided into control group, MDZ group, pcDNA3.1 group, and pcDNA3.1-YAP/TAZ group, and transfection efficiency was detected using qRT-PCR method; EdU staining, flow cytometry, and Transwell cells were applied to detect cell proliferation, apoptosis, and invasion, respectively; Western blot method was applied to detect the expression of proliferating cell nuclear antigen (PCNA), Bcl-2 associated X protein (Bax), N-cadherin, E-cadherin, and YAP, TAZ proteins; OS nude mouse model was constructed, and grouped into a control group and an MDZ group, immunohistochemical methods were applied to detect the expression of Ki-67, YAP, and TAZ proteins in transplanted tumor tissue, and TUNEL staining was applied to detect apoptosis. Results: The expression levels of YAP and TAZ mRNA and protein in OS cell lines were obviously increased (P<0.05); MDZ obviously reduced the expression of PCNA, N-cadherin, YAP, and TAZ, inhibitd the proliferation and invasion of MG63 and U20S cells, increased the expression of Bax and E-cadherin, and promoted cell apoptosis. Overexpression of YAP/TAZ was able to reverse the inhibitory effects of MDZ on the proliferation and invasion of MG63 and U20S cells, and promote the apoptosis of MG63 and U20S cells (P<0.05); in vivo experiments showed that MDZ obviously reduced the mass and volume of transplanted tumors, and the expression levels of Ki-67, YAP, and TAZ, promoted apoptosis of transplanted tumor cells (P<0.05). Conclusion: MDZ may inhibit the YAP/TAZ signaling pathway, inhibit the proliferation and invasion of OS cells, and promote apoptosis.

Key words： Midazolam YAP/TAZ Osteosarcoma Proliferation Apoptosis Invasion

基金资助:湖北省卫生健康委科研项目,(编号:WJ2023F090)

通讯作者: 涂梦佳

引用本文:

奥婷婷, 涂梦佳, 张熙. 咪达唑仑调节YAP/TAZ信号通路对骨肉瘤细胞增殖凋亡和侵袭的影响[J]. 河北医学, 2024, 30(1): 1-8.
AO Tingting, TU Mengjia, ZHANG Xi. Effects of Midazolam on Proliferation Apoptosis and Invasion of Osteosarcoma Cells by Regulating the YAP/TAZ Signaling Pathway. HeBei Med, 2024, 30(1): 1-8.

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http://www.hbyxzzs.cn/CN/10.3969/j.issn.1006-6233.2024.01.01 或 http://www.hbyxzzs.cn/CN/Y2024/V30/I1/1

[1] Chen C,Xie L,Ren T,et al.Immunotherapy for osteosarcoma:fundamental mechanism,rationale,and recent breakthroughs[J].Cancer Lett,2021,500(1):1-10.
[2] Dai L,Li S,Li X,Jiang B.Propofol inhibits the malignant development of osteosarcoma U2OS cells via AMPK/FΟΧO1-mediated autophagy[J].Oncol Lett,2022,24(3):310-317.
[3] Yan R,Jin S,Liu H,et al.Dexmedetomidine inhibits cell malignancy in osteosarcoma cells via miR-520a-3p-YOD1 interactome[J].Biochem Biophys Res Commun,2021,543(1):56-64.
[4] Kang J,Zheng Z,Li X,et al.Midazolam exhibits antitumour and enhances the efficiency of Anti-PD-1 immunotherapy in hepatocellular carcinoma[J].Cancer Cell Int,2022,22(1):312-325.
[5] 季英龙,周全军,冀国力.咪达唑仑抑制骨肉瘤细胞增殖、迁移并诱导其凋亡[J].局解手术学杂志,2023,32(7):586-591.
[6] Chen Y,Wang Y,Zhai Y,et al.Cinobufacini injection suppresses the proliferation of human osteosarcoma cells by inhibiting PIN1-YAP/TAZ signaling pathway[J].Front Pharmacol,2023,14(1):1081363-1081374.
[7] Li Y,Yang S,Yang S.Verteporfin inhibits the progression of spontaneous osteosarcoma caused by trp53 and Rb1 deficiency in ctsk-expressing cells via impeding hippo pathway[J].Cells,2022,11(8):1361-1372.
[8] 鲁飞,王强,谷淼.气体麻醉剂影响骨肉瘤细胞侵袭转移能力及化疗敏感性的分子机制[J].临床和实验医学杂志,2020,19(24):2614-2618.
[9] Wang Z,Song Y,Zhang H,et al.Local anesthetic levobupivacaine inhibits stemness of osteosarcoma cells by epigenetically repressing MAFB though reducing KAT5 expression[J].Aging (Albany NY),2022,14(6):2793-2804.
[10] 于鹏,尹天翔,罗彩云,等.盐酸罗哌卡因对骨肉瘤细胞增殖、侵袭、凋亡的影响及其机制[J].中国应用生理学杂志,2021,37(6):654-659,664.
[11] Zhang X,Han Z,Li Z,et al.Midazolam impedes lung carcinoma cell proliferation and migration via EGFR/MEK/ERK signaling pathway[J].Open Med (Wars),2023,18(1):20230730-20230740.
[12] Oshima Y,Sano M,Kajiwara I,et al.Midazolam exhibits antitumour and anti-inflammatory effects in a mouse model of pancreatic ductal adenocarcinoma[J].Br Anaesth,2022,128(4):679-690.
[13] Thompson BJ.YAP/TAZ:drivers of tumor growth,metastasis and Resistance to Therapy[J].Bioessays,2020,42(5):e1900162-e1900177.
[14] Chim LK,Williams IL,Bashor CJ,et al.Tumor-associated macrophages induce inflammation and drug resistance in a mechanically tunable engineered model of osteosarcoma[J].Biomaterials,2023,296(1):122076-122086.