miR383调控PRDX3表达及对人髓母细胞瘤增殖和凋亡影响的研究
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摘要
目的探讨muR383调控PRDX3表达及对人髓母细胞瘤增殖和凋亡的影响。方法新鲜组织样品来自于某医院2015年12月至2017年12月就诊的人髓母细胞瘤患者35例;另10例非肿瘤脑组织来自于该院病理科,作为对照组。人髓母瘤细胞系Daoy购自中国科学院细胞资源中心。采用Western blot法分别检测PRDX3在人髓母细胞瘤组织、Daoy细胞系、对照组脑组织三者中的表达情况。MTT法检测miR383转染细胞与对照组细胞增殖情况,采用流式细胞仪检测miR383转染细胞与对照组细胞凋亡情况。结果人髓母细胞瘤组织、Daoy细胞系、对照组脑组织三者中PRDX3的灰度值与β内参照计算比值,差异有统计学意义(P <0.05),表示三组细胞中PRDX3的表达水平有差异。人髓母细胞瘤组织中PRDX3的表达水平高于对照组脑组织,差异有统计学意义(P<0.05);Daoy细胞系中PRDX3的表达水平高于对照组脑组织,差异有统计学意义(P <0.05)。miR383转染组在转染48 h及72h后,PRDX3的表达水平低于Control组,差异有统计学意义(P <0.05);miR383转染组的Daoy细胞系增殖能力低于Control组水平,差异有统计学意义(P <0. 05);在第3天时,差异最为显著。miR383转染组细胞凋亡比例高于Control组,差异有统计学意义(P <0.05)。结论 PRDX3在人髓母细胞瘤组织、Daoy细胞系中存在过表达情况,而miR383可能通过下调PRDX3的表达水平来抑制Daoy细胞系的增殖,并促进其凋亡。
Objective To investigate the effects of miR383 on PRDX3 expression and how this regulation affects proliferation and apoptosis in human medulloblastoma. Methods Fresh tissue samples were collected from 35 patients with medulloblastoma in a hospital from 2015 to2017, and 10 other non-tumor brain tissues from the pathology department of our hospital were used as the control group. The human myeloma cell line Daoy was purchased from the cell resources center of Chinese Academy of Sciences. Western blot was used to detect the expression of PRDX3 in three human bone marrow blastoma tissues,Daoy cell lines and control brain tissues.MTT assay was used to detect the proliferation of miR383 transfected cells and control cells. Flowcytometry was used to detect the apoptosis of miR383 transfected cells and control groups. Results The gray values of PRDX3 among human marrow blastoma tissue, Daoy cell line and control group were compared by dividing the beta internal reference, and the difference was significantly different(P < 0.05),indicating that the expression levels of PRDX3 in the three groups were different. The expression of PRDX3 in the tissue of human marrow blastoma was higher than that of the control group(P <0.05),and the expression of PRDX3 in the Daoy cell line was higher than that of the control group(P <0.05). The expression level of PRDX3 in the transfected group was lower than that of control group after 48 hours and 72 hours of transfection(P <0.05). The proliferation ability of Daoy cell lines in miR383 transfected group was lower than that of the control group,(P <0. 05). The difference was the most significant at the day 3. The percentage of apoptotic cells in miR383 transfection group was higher than that in the control group(P <0.05). Conclusion PRDX3 is overexpressed in human marrow blastoma tissue and Daoy cell line,and miR383 may inhibit the proliferation of Daoy cell lines and promote the apoptosis by down-regulating the expression level of PRDX3.
Objective To investigate the effects of miR383 on PRDX3 expression and how this regulation affects proliferation and apoptosis in human medulloblastoma. Methods Fresh tissue samples were collected from 35 patients with medulloblastoma in a hospital from 2015 to2017, and 10 other non-tumor brain tissues from the pathology department of our hospital were used as the control group. The human myeloma cell line Daoy was purchased from the cell resources center of Chinese Academy of Sciences. Western blot was used to detect the expression of PRDX3 in three human bone marrow blastoma tissues,Daoy cell lines and control brain tissues.MTT assay was used to detect the proliferation of miR383 transfected cells and control cells. Flowcytometry was used to detect the apoptosis of miR383 transfected cells and control groups. Results The gray values of PRDX3 among human marrow blastoma tissue, Daoy cell line and control group were compared by dividing the beta internal reference, and the difference was significantly different(P < 0.05),indicating that the expression levels of PRDX3 in the three groups were different. The expression of PRDX3 in the tissue of human marrow blastoma was higher than that of the control group(P <0.05),and the expression of PRDX3 in the Daoy cell line was higher than that of the control group(P <0.05). The expression level of PRDX3 in the transfected group was lower than that of control group after 48 hours and 72 hours of transfection(P <0.05). The proliferation ability of Daoy cell lines in miR383 transfected group was lower than that of the control group,(P <0. 05). The difference was the most significant at the day 3. The percentage of apoptotic cells in miR383 transfection group was higher than that in the control group(P <0.05). Conclusion PRDX3 is overexpressed in human marrow blastoma tissue and Daoy cell line,and miR383 may inhibit the proliferation of Daoy cell lines and promote the apoptosis by down-regulating the expression level of PRDX3.
引文
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[13]王晓玫,张石芬,成志强,等.microRNA383调节人髓母细胞瘤Daoy细胞系中PRDX3的表达[J].中华病理学杂志,2012,41(8):547-552,
[14]陈锡彬,许哲,王乐浩,等.PRDX3在前列腺癌组织中表达的临床意义[J].广州医药,2014,45(5):1-3.
[15]丁妍,梁冰锋,张玲玲,等.食管鱗状细胞癌中PRDX3的表达及与其临床意义[J].临床合理用药杂志,2016,9(11):9-11.
[16] LI K K,PANG J C,LAU K M.et al. MiR-383 is downregulated in medulloblastoma and targets peroxiredoxin 3(PRDX3)[J]. Brain Pathol,2013,23(4):413-425.
[17] WHITAKER H C, PATEL D, HO WAT W J, et al. Peroxiredoxin-3is overexpressed in prostate cancer and promotes cancer cell survival by protecting cells from oxidative stress[J]. Br J Cancer,2013,109(4):983-993.
[18] WANG X M, ZHANG S F, CHENG Z Q, et al. MicroRNA383regulates expression of PRDX3 in human medulloblastomas[J].Chin J Pathol,2012,41(8):547-552.
[19] SHEN C,NATHAN C. Nonredundant antioxidant defense by multiple two-cysteine peroxiredoxins in human prostate cancer cells[J]. Mol Med,2002,8(2):95-102.
[20] FERRETTI E, De SMAELE E, MIELE E, et al. Concerted microRNA control of Hedgehog signalling in cerebellar neuronal progenitor and tumour cells[J]. EMBO J, 2014, 27(19):2616-2627.
[2]朱琳,章婧文,王慧,等.髓母细胞瘤手术联合放疗及放化疗病例的回顾性分析[J].中华神经医学杂志,2015, 14(3):239-243.
[3]田猛,方芳,李蹊,等.髓母细胞瘤中相关信号通路与靶向抑制剂研究进展[J].华西医学,2013,28(4):623-626.
[4] HU W,DANG X B, WANG G, et al. Peroxiredoxin-3 attenuates traumatic neuronal injury through preservation of mitochondrial function[J]. Neurochem Int,2018,114:120-126.
[5] BYUN J M, KIM S S, KIM K T, et al. Overexpression of peroxiredoxin-3 and-5 is a potential biomarker for prognosis in endometrial cancer[J]. Oncol Lett,2018,15(4):5111-5118.
[6]陈锡彬.PRDX3在前列腺癌组织中表达的临床意义及其对前列腺癌的保护作用[D].广州医科大学,2012.
[7] BASU A,BANERJEE H,ROJAS H,et al. Differential expression of peroxiredoxins in prostate cancer:Consistent upregulation of PRDX3and PRDX4[J].Prostate,2011,71(7):755-765.
[8]王凯,张姝,施露,等.2016年世界卫生组织中枢神经系统肿瘤分类概述[J].磁共振成像,2016,7(12):881-896.
[9]徐庆福.miRNA-22调控髓母细胞瘤增殖与凋亡的分子机理研究[D].第三军医大学,2014.
[10]许州,陈谦学,李明昌,等.胶质瘤中miR-383与其靶向circ_001634相互调控机制及作用研究[J].临床外科杂志,2016,24(11).844-847.
[11]周争光,汪蕊,李玉梅,等.mir-130a-3p及smad4在肝细胞癌组织中的表达及临床意义[J].安徽医科大学学报,2017,52(3):383-387.
[12]王晓玫,张石芬,成志强,等.上调microRNA-383对人髓母细胞瘤D341细胞系中PRDX3表达的影响[J].临床与实验病理学杂志,2012,28(4):422-428.
[13]王晓玫,张石芬,成志强,等.microRNA383调节人髓母细胞瘤Daoy细胞系中PRDX3的表达[J].中华病理学杂志,2012,41(8):547-552,
[14]陈锡彬,许哲,王乐浩,等.PRDX3在前列腺癌组织中表达的临床意义[J].广州医药,2014,45(5):1-3.
[15]丁妍,梁冰锋,张玲玲,等.食管鱗状细胞癌中PRDX3的表达及与其临床意义[J].临床合理用药杂志,2016,9(11):9-11.
[16] LI K K,PANG J C,LAU K M.et al. MiR-383 is downregulated in medulloblastoma and targets peroxiredoxin 3(PRDX3)[J]. Brain Pathol,2013,23(4):413-425.
[17] WHITAKER H C, PATEL D, HO WAT W J, et al. Peroxiredoxin-3is overexpressed in prostate cancer and promotes cancer cell survival by protecting cells from oxidative stress[J]. Br J Cancer,2013,109(4):983-993.
[18] WANG X M, ZHANG S F, CHENG Z Q, et al. MicroRNA383regulates expression of PRDX3 in human medulloblastomas[J].Chin J Pathol,2012,41(8):547-552.
[19] SHEN C,NATHAN C. Nonredundant antioxidant defense by multiple two-cysteine peroxiredoxins in human prostate cancer cells[J]. Mol Med,2002,8(2):95-102.
[20] FERRETTI E, De SMAELE E, MIELE E, et al. Concerted microRNA control of Hedgehog signalling in cerebellar neuronal progenitor and tumour cells[J]. EMBO J, 2014, 27(19):2616-2627.