瑞芬太尼通过miR-206/GOLPH3调控胃癌细胞增殖和凋亡的实验研究
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摘要
背景瑞芬太尼为临床上常用的麻醉药,近几年其新功效不断被发现,尤其是抗癌功能.但瑞芬太尼在胃癌(gastric cancer, GC)中的作用及机制尚未清楚.目的探讨瑞芬太尼对A G S、M K N-45人G C细胞中mi R-206、GOLPH3表达及细胞增殖、凋亡的影响.方法以40 nmol/L瑞芬太尼干预AGS、MKN-45细胞, q RTPCR、Westernblot、MTT和流式细胞仪分别检测细胞中mi R-206和GOLPH3表达及细胞活力、凋亡.在AGS、MKN-45细胞中过表达miR-206或敲减GOLPH3,MTT和流式细胞仪检测细胞活力和凋亡.Targetscan在线预测、双荧光素酶报告基因实验和Westernblot实验验证miR-206和GOLPH3的靶向关系.将miR-206 inhibitors或pcDNA-GOLPH3转染至AGS、MKN-45细胞并以40nmol/L瑞芬太尼进行处理,检测细胞活力和凋亡.结果瑞芬太尼干预的AGS、MKN-45细胞中miR-206高表达而GOLPH3低表达,细胞活力降低而凋亡率升高.过表达mi R-206或敲减GOLPH3,细胞活力下降、凋亡率升高. Targetscan在线预测、双荧光素酶报告基因实验和Western blot实验结果表明, miR-206可靶向调控GOLPH3蛋白表达.下调miR-206或过表达GOLPH3能够逆转瑞芬太尼对AGS、MKN-45细胞增殖的抑制和凋亡的促进作用.结论瑞芬太尼能够通过调节miR-206和GOLPH3表达抑制AGS、MKN-45细胞增殖并诱导凋亡.
BACKGROUND Remifentanil is a commonly used anesthetic inclinical practice. In recent years, its new efficacy has been continuously discovered, especially its anticancer function. However, the role and mechanism of remifentanil in gastric cancer(GC) are still not clear.AIM To investigate the effect of remifentanil on the expression of miR-206, GOLPH3, cell proliferation, and apoptosis in human GC cell lines AGS and MKN-45. METHODS The expression of miR-206 and GOLPH3 and the viability and apoptosis of AGS and MKN-45 cells after treatment with 40 nmol/L remifentanil were detected by qRT-PCR, Western blot, MTT assay, and flow cytometry, respectively. Cell viability and apoptosis of AGS and MKN-45 cells with overexpression of miR-206 or knockdown of GOLPH3 were detected by MTT assay and flow cytometry, respectively. The targeting relationship between miR-206 and GOLPH3 was verified by Targetscan online prediction, dual-luciferase assay, and Western blot. After transfection with miR-206 inhibitor or pc DNA-GOLPH3, AGS and MKN-45 cells were treated with 40 nmol/L remifentanil and then detected for cell viability and apoptosis. RESULTS After treatment with remifentanil, the expression of miR-206 and apoptosis rate were increased while the expression of GOLPH3 and cell viability were decreased in AGS and MKN-45 cells. Cell viability was decreased and apoptotic rate was increased in AGS and MKN-45 cells after overexpression of mi R-206 or knockdown of GOLPH3. The results of Targetscan online prediction, dual-luciferase assay, and Western blot indicted that miR-206 could regulate the expression of GOLPH3 protein. Down-regulation of mi R-206 or overexpression of GOLPH3 could reverse the inhibition of proliferationand apoptosis of AGS and MKN-45 cells by remifentanil. CONCLUSION Remifentanil could inhibit the proliferation and induce apoptosis of AGS and MKN-45 cells by regulating the expression of miR-206 and GOLPH3.
BACKGROUND Remifentanil is a commonly used anesthetic inclinical practice. In recent years, its new efficacy has been continuously discovered, especially its anticancer function. However, the role and mechanism of remifentanil in gastric cancer(GC) are still not clear.AIM To investigate the effect of remifentanil on the expression of miR-206, GOLPH3, cell proliferation, and apoptosis in human GC cell lines AGS and MKN-45. METHODS The expression of miR-206 and GOLPH3 and the viability and apoptosis of AGS and MKN-45 cells after treatment with 40 nmol/L remifentanil were detected by qRT-PCR, Western blot, MTT assay, and flow cytometry, respectively. Cell viability and apoptosis of AGS and MKN-45 cells with overexpression of miR-206 or knockdown of GOLPH3 were detected by MTT assay and flow cytometry, respectively. The targeting relationship between miR-206 and GOLPH3 was verified by Targetscan online prediction, dual-luciferase assay, and Western blot. After transfection with miR-206 inhibitor or pc DNA-GOLPH3, AGS and MKN-45 cells were treated with 40 nmol/L remifentanil and then detected for cell viability and apoptosis. RESULTS After treatment with remifentanil, the expression of miR-206 and apoptosis rate were increased while the expression of GOLPH3 and cell viability were decreased in AGS and MKN-45 cells. Cell viability was decreased and apoptotic rate was increased in AGS and MKN-45 cells after overexpression of mi R-206 or knockdown of GOLPH3. The results of Targetscan online prediction, dual-luciferase assay, and Western blot indicted that miR-206 could regulate the expression of GOLPH3 protein. Down-regulation of mi R-206 or overexpression of GOLPH3 could reverse the inhibition of proliferationand apoptosis of AGS and MKN-45 cells by remifentanil. CONCLUSION Remifentanil could inhibit the proliferation and induce apoptosis of AGS and MKN-45 cells by regulating the expression of miR-206 and GOLPH3.
引文
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7 Glass PS,Gan TJ,Howell S.A review of the pharmacokinetics and pharmacodynamics of remifentanil.Anesth Analg 1999;89:S7-14[PMID:10511072]
8唐优仕,王龙,马浩文,夏中元,赵博.瑞芬太尼对胰腺癌Bx PC-3细胞增殖、凋亡的影响及其机制.中国热带医学2017;17:1193-1197[DOI:10.13604/j.cnki.46-1064/r.2017.12.07]
9赵莉,王志红,李学斌,徐桂萍.瑞芬太尼对人结肠癌COL0205细胞增殖及凋亡的影响2017;38:401-403[DOI:10.3760/cma.j.issn.1673-4378.2017.05.004]
10刘思同,翟志超,左明明,刘金锋.非阿片类镇痛药物在癌痛中的应用.中国疼痛医学杂志2016;22:223-227[DOI:10.3969/j.issn.1006-9852.2016.03.014]
11周成茂,阮林.阿片类受体在蛋白激酶B通路相关肿瘤复发中的作用.国际麻醉学与复苏杂志2015;36:479-480[DOI:10.3760/cma.j.issn.1673-4378.2015.05.023]
12孟俊青.芬太尼和瑞芬太尼对人肺癌细胞A549细胞活力的影响.河北医科大学,2012[DOI:10.7666/d.y2105588]
13 Rupaimoole R,Slack FJ.Micro RNA therapeutics:towards a new era for the management of cancer and other diseases.Nat Rev Drug Discov 2017;16:203-222[PMID:28209991 DOI:10.1038/nrd.2016.246]
14 Li YY,Shao JP,Zhang SP,Xing GQ,Liu HJ.miR-519d-3p Inhibits Cell Proliferation and Invasion of Gastric Cancer by Downregulating B-Cell Lymphoma 6.Cytogenet Genome Res2018;154:12-19[PMID:29510377 DOI:10.1159/000487372]
15 Wang Y,Zhang H,Ge S,Fan Q,Zhou L,Li H,Bai M,Ning T,Liu R,Wang X,Deng T,Zhang L,Ying G,Ba Y.Effects of miR-138-5p and miR-204-5p on the migration and proliferation of gastric cancer cells by targeting EGFR.Oncol Rep 2018;39:2624-2634[PMID:29693184 DOI:10.3892/or.2018.6389]
16张霖,廉姜芳,周建庆.MicroRNA-206功能的研究进展.中国细胞生物学学报2015;37:1046-1052[DOI:10.11844/cjcb.2015.07.0083]
17 Pan JY,Sun CC,Bi ZY,Chen ZL,Li SJ,Li QQ,Wang YX,Bi YY,Li DJ.miR-206/133b Cluster:A Weapon against Lung Cancer?Mol Ther Nucleic Acids 2017;8:442-449[PMID:28918043 DOI:10.1016/j.omtn.2017.06.002]
18 Pang C,Huang G,Luo K,Dong Y,He F,Du G,Xiao M,Cai W.mi R-206 inhibits the growth of hepatocellular carcinoma cells via targeting CDK9.Cancer Med 2017;6:2398-2409[PMID:28940993 DOI:10.1002/cam4.1188]
19 Wang T,Dong XM,Zhang FL,Zhang JR.miR-206 enhances nasopharyngeal carcinoma radiosensitivity by targeting IGF1.Kaohsiung J Med Sci 2017;33:427-432[PMID:28865599 DOI:10.1016/j.kjms.2017.05.015]
20 Wang R,Zhang T,Yang Z,Jiang C,Seng J.Long non-coding RNA FTH1P3 activates paclitaxel resistance in breast cancer through miR-206/ABCB1.J Cell Mol Med 2018;22:4068-4075[PMID:29971911 DOI:10.1111/jcmm.13679]
21 Gong YC,Ren GL,Liu B,Li F,Zhao HP,Chen JB,Li YP,Yu HH.mi R-206 inhibits cancer initiating cells by targeting EHF in gastric cancer.Oncol Rep 2017;38:1688-1694[PMID:28714026 DOI:10.3892/or.2017.5794]
22 Rizzo R,Parashuraman S,D’Angelo G,Luini A.GOLPH3 and oncogenesis:What is the molecular link?Tissue Cell 2017;49:170-174[PMID:27378035 DOI:10.1016/j.tice.2016.06.008]
23 Zhou B,Wang G,Gao S,Chen Y,Jin C,Wang Z,Yang Y,Ma Z,Zhang W,Feng X.Expression of GOLPH3 protein in colon cancer tissues and its association with the prognosis of patients.Oncol Lett 2016;12:3936-3940[PMID:27895752 DOI:10.3892/ol.2016.5215]
24 Sun J,Yang X,Zhang R,Liu S,Gan X,Xi X,Zhang Z,Feng Y,Sun Y.GOLPH3 induces epithelial-mesenchymal transition via Wnt/β-catenin signaling pathway in epithelial ovarian cancer.Cancer Med 2017;6:834-844[PMID:28332316 DOI:10.1002/cam4.1040]
25 Wang JH,Yuan LJ,Liang RX,Liu ZG,Li BH,Wen ZS,Huang ST,Zheng M.GOLPH3 promotes cell proliferation and tumorigenicity in esophageal squamous cell carcinoma via mTOR and Wnt/β-catenin signal activation.Mol Med Rep 2017;16:7138-7144[PMID:28901498 DOI:10.3892/mmr.2017.7495]
26 Liu H,Wang X,Feng B,Tang L,Li W,Zheng X,Liu Y,Peng Y,Zheng G,He Q.Golgi phosphoprotein 3(GOLPH3)promotes hepatocellular carcinoma progression by activating m TOR signaling pathway.BMC Cancer 2018;18:661[PMID:29914442 DOI:10.1186/s12885-018-4458-7]
27 Liu Y,Sun Y,Zhao A.MicroRNA-134 suppresses cell proliferation in gastric cancer cells via targeting of GOLPH3.Oncol Rep 2017;37:2441-2448[PMID:28260021 DOI:10.3892/or.2017.5488]
2 Chen W,Zheng R,Baade PD,Zhang S,Zeng H,Bray F,Jemal A,Yu XQ,He J.Cancer statistics in China,2015.CACancer J Clin 2016;66:115-132[PMID:26808342 DOI:10.3322/caac.21338]
3 Quan Y,Huang A,Ye M,Xu M,Zhuang B,Zhang P,Yu B,Min Z.Comparison of laparoscopic versus open gastrectomy for advanced gastric cancer:an updated meta-analysis.Gastric Cancer 2016;19:939-950[PMID:26216579 DOI:10.1007/s10120-015-0516-x]
4 Yuan DD,Zhu ZX,Zhang X,Liu J.Targeted therapy for gastric cancer:Current status and future directions(Review).Oncol Rep 2016;35:1245-1254[PMID:26718131 DOI:10.3892/or.2015.4528]
5 Izuishi K,Mori H.Recent Strategies for Treating Stage IV Gastric Cancer:Roles of Palliative Gastrectomy,Chemotherapy,and Radiotherapy.J Gastrointestin Liver D i s 2016;25:87-94[P M I D:27014758 D O I:10.15403/jgld.2014.1121.251.rv2]
6 Digklia A,Wagner AD.Advanced gastric cancer:Current treatment landscape and future perspectives.World JGastroenterol 2016;22:2403-2414[PMID:26937129 DOI:10.3748/wjg.v22.i8.2403]
7 Glass PS,Gan TJ,Howell S.A review of the pharmacokinetics and pharmacodynamics of remifentanil.Anesth Analg 1999;89:S7-14[PMID:10511072]
8唐优仕,王龙,马浩文,夏中元,赵博.瑞芬太尼对胰腺癌Bx PC-3细胞增殖、凋亡的影响及其机制.中国热带医学2017;17:1193-1197[DOI:10.13604/j.cnki.46-1064/r.2017.12.07]
9赵莉,王志红,李学斌,徐桂萍.瑞芬太尼对人结肠癌COL0205细胞增殖及凋亡的影响2017;38:401-403[DOI:10.3760/cma.j.issn.1673-4378.2017.05.004]
10刘思同,翟志超,左明明,刘金锋.非阿片类镇痛药物在癌痛中的应用.中国疼痛医学杂志2016;22:223-227[DOI:10.3969/j.issn.1006-9852.2016.03.014]
11周成茂,阮林.阿片类受体在蛋白激酶B通路相关肿瘤复发中的作用.国际麻醉学与复苏杂志2015;36:479-480[DOI:10.3760/cma.j.issn.1673-4378.2015.05.023]
12孟俊青.芬太尼和瑞芬太尼对人肺癌细胞A549细胞活力的影响.河北医科大学,2012[DOI:10.7666/d.y2105588]
13 Rupaimoole R,Slack FJ.Micro RNA therapeutics:towards a new era for the management of cancer and other diseases.Nat Rev Drug Discov 2017;16:203-222[PMID:28209991 DOI:10.1038/nrd.2016.246]
14 Li YY,Shao JP,Zhang SP,Xing GQ,Liu HJ.miR-519d-3p Inhibits Cell Proliferation and Invasion of Gastric Cancer by Downregulating B-Cell Lymphoma 6.Cytogenet Genome Res2018;154:12-19[PMID:29510377 DOI:10.1159/000487372]
15 Wang Y,Zhang H,Ge S,Fan Q,Zhou L,Li H,Bai M,Ning T,Liu R,Wang X,Deng T,Zhang L,Ying G,Ba Y.Effects of miR-138-5p and miR-204-5p on the migration and proliferation of gastric cancer cells by targeting EGFR.Oncol Rep 2018;39:2624-2634[PMID:29693184 DOI:10.3892/or.2018.6389]
16张霖,廉姜芳,周建庆.MicroRNA-206功能的研究进展.中国细胞生物学学报2015;37:1046-1052[DOI:10.11844/cjcb.2015.07.0083]
17 Pan JY,Sun CC,Bi ZY,Chen ZL,Li SJ,Li QQ,Wang YX,Bi YY,Li DJ.miR-206/133b Cluster:A Weapon against Lung Cancer?Mol Ther Nucleic Acids 2017;8:442-449[PMID:28918043 DOI:10.1016/j.omtn.2017.06.002]
18 Pang C,Huang G,Luo K,Dong Y,He F,Du G,Xiao M,Cai W.mi R-206 inhibits the growth of hepatocellular carcinoma cells via targeting CDK9.Cancer Med 2017;6:2398-2409[PMID:28940993 DOI:10.1002/cam4.1188]
19 Wang T,Dong XM,Zhang FL,Zhang JR.miR-206 enhances nasopharyngeal carcinoma radiosensitivity by targeting IGF1.Kaohsiung J Med Sci 2017;33:427-432[PMID:28865599 DOI:10.1016/j.kjms.2017.05.015]
20 Wang R,Zhang T,Yang Z,Jiang C,Seng J.Long non-coding RNA FTH1P3 activates paclitaxel resistance in breast cancer through miR-206/ABCB1.J Cell Mol Med 2018;22:4068-4075[PMID:29971911 DOI:10.1111/jcmm.13679]
21 Gong YC,Ren GL,Liu B,Li F,Zhao HP,Chen JB,Li YP,Yu HH.mi R-206 inhibits cancer initiating cells by targeting EHF in gastric cancer.Oncol Rep 2017;38:1688-1694[PMID:28714026 DOI:10.3892/or.2017.5794]
22 Rizzo R,Parashuraman S,D’Angelo G,Luini A.GOLPH3 and oncogenesis:What is the molecular link?Tissue Cell 2017;49:170-174[PMID:27378035 DOI:10.1016/j.tice.2016.06.008]
23 Zhou B,Wang G,Gao S,Chen Y,Jin C,Wang Z,Yang Y,Ma Z,Zhang W,Feng X.Expression of GOLPH3 protein in colon cancer tissues and its association with the prognosis of patients.Oncol Lett 2016;12:3936-3940[PMID:27895752 DOI:10.3892/ol.2016.5215]
24 Sun J,Yang X,Zhang R,Liu S,Gan X,Xi X,Zhang Z,Feng Y,Sun Y.GOLPH3 induces epithelial-mesenchymal transition via Wnt/β-catenin signaling pathway in epithelial ovarian cancer.Cancer Med 2017;6:834-844[PMID:28332316 DOI:10.1002/cam4.1040]
25 Wang JH,Yuan LJ,Liang RX,Liu ZG,Li BH,Wen ZS,Huang ST,Zheng M.GOLPH3 promotes cell proliferation and tumorigenicity in esophageal squamous cell carcinoma via mTOR and Wnt/β-catenin signal activation.Mol Med Rep 2017;16:7138-7144[PMID:28901498 DOI:10.3892/mmr.2017.7495]
26 Liu H,Wang X,Feng B,Tang L,Li W,Zheng X,Liu Y,Peng Y,Zheng G,He Q.Golgi phosphoprotein 3(GOLPH3)promotes hepatocellular carcinoma progression by activating m TOR signaling pathway.BMC Cancer 2018;18:661[PMID:29914442 DOI:10.1186/s12885-018-4458-7]
27 Liu Y,Sun Y,Zhao A.MicroRNA-134 suppresses cell proliferation in gastric cancer cells via targeting of GOLPH3.Oncol Rep 2017;37:2441-2448[PMID:28260021 DOI:10.3892/or.2017.5488]