一水草酸钙晶体刺激肾近端小管细胞后miRNA表达谱的变化
伯豪生物
2451
华中科技大学同济医学院泌尿外科研究所的研究人员证实,在一水草酸钙刺激肾近端小管细胞后,miRNA的表达谱发生了变化,这将暗示着miRNA也许在结石形成中发挥作用。
背景介绍
结石形成的原因至今尚未探知,而一水草酸钙(COM)是结石中主要成分,本文通过COM刺激HK-2细胞后的miRNA表达谱的变化,初步探讨miRNA在刺激过程中的作用。
文章亮点
通过LDH和DAPI染色方法测定最适宜浓度,通过miRNA和mRNA芯片测定表达谱变化,通过real-time PCR鉴定表达谱芯片,通过GO和PATHWAY分析预测生物学行为(课题相关Agilent human miRNA microarrays技术服务由上海伯豪生物技术有限公司提供)。
结果分析
1mmol/L的COM晶体24h刺激完后有25条miRNAs表达超过了1.5倍,其中16条是上升的,9条下降。
Dysregulated miRNAs after COM crystals exposure
通过GO和PATHWAY分析这些变化的miRNA与细胞凋亡、代谢反应、信号传递等生物学效应有关, 提示miRNA在结石形成过程中可能存在作用。
Analysis of biological processes of the predicted miRNAs targets by GOstat
原文出处
Wang BH, Wu BL, Liu J, Yao WM, Xia D, Li L, Chen ZQ, Ye ZQ, Yu X. Analysis of Altered MicroRNA Expression Profiles in Proximal Renal Tubular Cells in Response to Calcium
Oxalate Monohydrate Crystal Adhesion: Implications for Kidney Stone Disease. Plos One.2014,9(7): e101306.
背景介绍
结石形成的原因至今尚未探知,而一水草酸钙(COM)是结石中主要成分,本文通过COM刺激HK-2细胞后的miRNA表达谱的变化,初步探讨miRNA在刺激过程中的作用。
文章亮点
通过LDH和DAPI染色方法测定最适宜浓度,通过miRNA和mRNA芯片测定表达谱变化,通过real-time PCR鉴定表达谱芯片,通过GO和PATHWAY分析预测生物学行为(课题相关Agilent human miRNA microarrays技术服务由上海伯豪生物技术有限公司提供)。
结果分析
1mmol/L的COM晶体24h刺激完后有25条miRNAs表达超过了1.5倍,其中16条是上升的,9条下降。
Dysregulated miRNAs after COM crystals exposure
Probe_ID | foldchange | Probe_ID | Foldchange |
Up-regulated | Down-regulated | ||
hsa-miR-371-5p | 3.069941 | hsa-miR-42undefined | 0.069018 |
hsa-miR-3125 | 2.71054 | hsv1-miR-H6-3p | 0.071335 |
hsa-miR-638 | 2.682127 | hsv1-miR-Hundefined | 0.078529 |
hsa-miR-3195 | 2.251545 | hsa-miR-933 | 0.09623093 |
ebv-miR-BART13 | 2.155447 | hsa-miR-19undefined | 0.303036 |
hsa-miR-135undefined | 2.078597 | hsa-let-7f-undefined | 0.366442 |
hsa-miR-1274b | 2.004063 | hsa-miR-4284 | 0.393521 |
hsa-miR-497 | 1.918096 | hsa-miR-4313 | 0.485941 |
hsa-miR-1181 | 1.829789 | hsa-let-7undefined | 0.48814 |
hsa-miR-195 | 1.706755 | ||
hsa-miR-1260b | 1.693521 | ||
hsa-miR-2undefined | 1.688719 | ||
hsa-miR-642b | 1.644018 | ||
hsa-miR-1260 | 1.585072 | ||
hsv1-miR-H8 | 1.532887 | ||
hsa-miR-1274a | 1.505559 |
通过GO和PATHWAY分析这些变化的miRNA与细胞凋亡、代谢反应、信号传递等生物学效应有关, 提示miRNA在结石形成过程中可能存在作用。
Analysis of biological processes of the predicted miRNAs targets by GOstat
miRNAs | GO process | Target genes | Count | % | P-value |
hsa-miR-497 | GO:0043549:regulation of kinase activity | PPP2R1A, MADD, PTPLAD1, DRD5, PDCD4, CDC25A, CDC25B, CBLC, TARBP2, PRKAR2A, APP, TSPYL2, DUSP9, HTR2A | 14 | 0.3688 | 0.04444 |
GO:0007249:I-kappaB kinase/NF-kappaB cascade | PTPLAD1, SNIP1, TLR1, IKBKB, TNIP2 | 5 | 0.1317 | 0.04600 | |
GO:0031980:mitochondrial lumen | GLS2, MRPL40, PDPR, FECH, HARS2, TMLHE, CARS2, YARS2, AK3L1, LRPPRC, KARS | 11 | 0.2898 | 0.02395 | |
GO:0005739:mitochondrion | MRPL40, ACOX1, D2HGDH, CCDC19, NLRX1, AK3L1, KCNJ11, KARS, GLS2, GPX1, GSR, HTRA2, CISD3, SLC25A22, HARS2, SLC25A2, BDH2, AATK, PDPR, PPP2R1A, TRMU, FECH, NOL3, MRPS2, MMAB, RAB40AL, UCP2, TMLHE, SLC25A35, CARS2, YARS2, ALKBH3, LRPPRC | 33 | 0.8693 | 0.03916 | |
hsa-miR-638 | GO:0009611:response to wounding | CCL11, MYF6, GPX1, TNFRSF1B, F10, MGLL, HOXB13, IL11 | 8 | 0.9335 | 0.00599 |
GO:0010740:positive regulation of protein kinase cascade | GPX1, F10, BST2, IL11 | 4 | 0.4667 | 0.03180 | |
GO:0031099:regeneration | MYF6, GPX1, NR4A3 | 3 | 0.3501 | 0.03308 | |
hsa-miR-1260 | GO:0051252:regulation of RNA metabolic process | ZNF583, SNIP1, PDX1, ZNF253, SKAP1, IL11, APP, GFI1B, RAB26, SLA2, OTX1, ARID5A, SOX12, HDAC10, FOSB, NPAS4, SPEN, UBN1, ZNF192, ZNF786, MED8, SERBP1, TGIF2, CUX1, TFAP2E, MAPRE3, MED1, CARHSP1, FOXM1, HOXB13, FHL2, ZNF367, NEDD8, ZNF177, SUFU, ZNF615, ZNF169, TNFRSF1B, TSC22D2, TCEA3, HEXIM2, LHX1, OVOL1, GATAD2B, ZNF426, ZNF701, FOXD4, SIM2, CHD3, MYF6, ESRRA, ZNF565, CREBZF, TAF8, NR4A1, ZNF221, NR4A3, ZNF763, RFX3, MESP2, NCOR2 | 61 | 1.5204 | 0.00323 |
GO:0045449:regulation of transcription | ZNF583, SNIP1, PDX1, ZNF253, PDCD4, CBX7, SKAP1, IL11, ARHGAP22, BZW1, APP, LBH, GFI1B, MED29, RAB26, SSBP2, POGZ, OTX1, SLA2, SOX12, ARID5A, HDAC10, FOSB, HDAC11, SPEN, NPAS4, UBN1, CCNL2, ZNF192, CCR6, PRDM4, ZNF786, MED8, PARP14, ZNF384, ERN2, TGIF2, CUX1, TFAP2E, MAPRE3, MED1, CARHSP1, FOXM1, TRIB3, HOXB13, FHL2, ZNF367, NEDD8, ZNF177, ZNF615, SUFU, ZNF169, TSC22D2, TCEA3, HEXIM2, LHX1, OVOL1, GATAD2B, ZNF426, RNF10, ZNF701, FOXD4, CHD3, SIM2, MYF6, SSRP1, ESRRA, ZBTB46, ZNF565, CREBZF, TAF8, NR4A1, ZNF221, NR4A3, ZNF629, PHF19, ZNF763, RFX3, IKBKB, MESP2, NCOR2 | 80 | 1.9940 | 0.00686 | |
GO:0043167:ion binding | SLC13A5, ZNF583, ZNRF1, ZNF253, TTL, APOBEC3C, APOBEC3D, MAP3K5, APP, CISD3, CDH23, RET, CAPNS2, FECH, PDXK, CRTAC1, BSN, RNASEH2A, WDR92, ZNF192, PRDM4, ZNF786, EIF2S2, ZNF384, UNC13A, NEK3, ZNF615, KCNJ1, RPS27, ZNF428, LHX1, OVOL1, RNF168, RNF10, ZNF426, STK19, GABRP, GIT1, NPLOC4, ESRRA, ZBTB46, B4GALT2, SETDB2, NR4A1, ZNF221, NR4A3, ACACB, ATP13A3, CYB561, PRPSAP2, ZNF629, TRIM56, KCNJ9, TMLHE, ZNF385D, SPTA1, TRAFD1, GLRA1, MLPH, GLRA3, CYP11B2, PDIA4, KCNJ11, KCNIP1, USP19, SMAP2, GFI1B, STAC2, TRIM9, DBR1, NSF, NT5C, POGZ, MICAL3, RPH3AL, CYB5B, ARL3, MYRIP, MAP3K15, ZNF691, TAF15, ERN2, CA4, CLCNKA, ZNF367, FHL2, EGLN2, ZNF177, BEST4, ZNF169, CYB561D1, TCEA3, CALML3, MMACHC, PRRG2, RASGRP2, DTNB, ENO2, MT1E, GATAD2B, GALNT11, ZNF701, CHD3, MT1M, NOS1, ZNF565, TRIM29, TRIM25, CYP4F8, RPS6KA3, FBLN1, RASSF5, PHF19, CDH15, ARSA, SCN4B, SUMF2, ZNF763, ALKBH3, RNF40 | 120 | 2.9910 | 0.03602 | |
hsa-miR-1260b | GO:0051252:regulation of RNA metabolic process | ZNF583, SNIP1, PDX1, ZNF253, SKAP1, IL11, APP, GFI1B, RAB26, SLA2, OTX1, ARID5A, SOX12, HDAC10, FOSB, NPAS4, SPEN, UBN1, ZNF192, ZNF786, MED8, SERBP1, TGIF2, CUX1, TFAP2E, MAPRE3, MED1, CARHSP1, FOXM1, HOXB13, FHL2, ZNF367, NEDD8, ZNF177, SUFU, ZNF615, ZNF169, TNFRSF1B, TSC22D2, TCEA3, HEXIM2, LHX1, OVOL1, GATAD2B, ZNF426, ZNF701, FOXD4, SIM2, CHD3, MYF6, ESRRA, ZNF565, CREBZF, TAF8, NR4A1, ZNF221, NR4A3, ZNF763, RFX3, MESP2, NCOR2 | 61 | 1.5204 | 0.00323 |
GO:0007242:intracellular signaling cascade | FGFR1, CARHSP1, RAB9A, RAB3D, PTPLAD1, DRD5, SNIP1, MAPKAPK3, FHL2, EGLN2, PDX1, RAB42, MAP3K5, DIRAS1, PRKAR2A, KISS1R, STAC2, RASGRP2, RAB26, TNIP2, LTB, GDI1, RET, C5AR1, PIK3C2B, SLA2, STMN4, NFKBIL1, DEPDC5, ARL3, RASL11B, RASSF5, RPS6KA3, RAB37, CNIH3, BBC3, SGSM3, RAB17, ASB1, IKBKB, UNC13A, MED1 | 42 | 1.0469 | 0.01837 | |
hsa-miR-3125 | GO:0006350:transcription | RNASEK, FOXM1, CBX4, TTLL5, ZNF367, ZNF177, CBX7, ZNF615, DPF1, ZNF169, GFI1B, OVOL1, GATAD2B, TWIST2, FOXD4, CHD3, ZNF548, ZBTB46, BRF2, TRIM29, SOX12, CDK9, DACH1, NR4A3, SPEN, HDAC11, HMGA1, NRIP2, CCR6, PRDM4, MED8, PARP14, ZNF384, RFX3, LRPPRC, PUF60, MED1 | 37 | 1.9004 | 0.01354 |
hsa-miR-3195 | GO:0002064:epithelial cell development | AGPAT6, HOXB13 | 2 | 0.9009 | 0.02456 |
GO:0004672:protein kinase activity | FGFR1, LMTK2, CDK9, IKBKB | 4 | 1.8018 | 0.04223 | |
hsa-miR-4284 | GO:0008629:induction of apoptosis by intracellular signals | CUL3, CUL2, BBC3, CIDEA | 4 | 0.1980 | 0.02512 |
hsa-miR-4313 | GO:0045449:regulation of transcription | CARHSP1, ZNF583, GDF7, HOXB13, CBX7, PDCD4, IL11, DPF1, CCNE1, TSPYL2, MED29, OVOL1, TWIST2, ESRRA, ZBTB46, POGZ, BRF2, ARID5A, HDAC11, DACH1, ZNF497, HMGA1, CCNL2, ZNF629, TARBP2, PRDM4, MED8, PARP14, ZNF384, ZNF763, TGIF2, JAK3, CUX1, MAPRE3, MESP2, LRPPRC, NCOR2 | 37 | 2.2534 | 0.02350 |
原文出处
Wang BH, Wu BL, Liu J, Yao WM, Xia D, Li L, Chen ZQ, Ye ZQ, Yu X. Analysis of Altered MicroRNA Expression Profiles in Proximal Renal Tubular Cells in Response to Calcium
Oxalate Monohydrate Crystal Adhesion: Implications for Kidney Stone Disease. Plos One.2014,9(7): e101306.