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一水草酸鈣晶體刺激腎近端小管細胞后miRNA表達譜的變化
關鍵詞： miRNA 表達譜 芯片來源： 伯豪生物
華中科技大學同濟醫學院泌尿外科研究所的研究人員證實，在一水草酸鈣刺激腎近端小管細胞后，miRNA的表達譜發生了變化，這將暗示著miRNA也許在結石形成中發揮作用。?背景介紹結石形成的原因至今尚未探知，而一水草酸鈣（COM）是結石中主要成分，本文通過COM刺激HK-2細胞后的miRNA表達譜的變化，初步探討miRNA在刺激過程中的作用。?文章亮點通過LDH和DAPI染色方法測定最適宜濃度，通過miRNA和mRNA芯片測定表達譜變化，通過real-time PCR鑒定表達譜芯片，通過GO和PATHWAY分析預測生物學行為(課題相關Agilent human miRNA microarrays技術服務由上海伯豪生物技術有限公司提供)。?結果分析1mmol/L的COM晶體24h刺激完后有25條miRNAs表達超過了1.5倍，其中16條是上升的，9條下降。?Dysregulated miRNAs after COM crystals exposure?
| Probe_ID | foldchange | Probe_ID | Foldchange |
| Up-regulated | ? | Down-regulated | ? |
| hsa-miR-371-5p | 3.069941 | hsa-miR-425* | 0.069018 |
| hsa-miR-3125 | 2.71054 | hsv1-miR-H6-3p | 0.071335 |
| hsa-miR-638 | 2.682127 | hsv1-miR-H7* | 0.078529 |
| hsa-miR-3195 | 2.251545 | hsa-miR-933 | 0.09623093 |
| ebv-miR-BART13 | 2.155447 | hsa-miR-191* | 0.303036 |
| hsa-miR-135a* | 2.078597 | hsa-let-7f-1* | 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-7b* | 0.48814 |
| hsa-miR-195 | 1.706755 | ? | ? |
| hsa-miR-1260b | 1.693521 | ? | ? |
| hsa-miR-21* | 1.688719 | ? | ? |
| hsa-miR-642b | 1.644018 | ? | ? |
| hsa-miR-1260 | 1.585072 | ? | ? |
| hsv1-miR-H8 | 1.532887 | ? | ? |
| hsa-miR-1274a | 1.505559 | ? | ? |
?通過GO和PATHWAY分析這些變化的miRNA與細胞凋亡、代謝反應、信號傳遞等生物學效應有關, 提示miRNA在結石形成過程中可能存在作用。?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 |
?原文出處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 CalciumOxalate Monohydrate Crystal Adhesion: Implications for Kidney Stone Disease. Plos One.2014,9(7): e101306.
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