利用微阵列芯片技术探究基因FOXQ1与大肠癌的关系

表1 FOXQ1敲低后, 上调倍数最大的20个基因

基因名称	倍性变化	mRNA编号	描述
GPA33	6.626402	NM_005814	Homo sapiens glycoprotein A33 (transmembrane) (GPA33)
DNMBP-AS1	6.349088	NR_024130	Homo sapiens DNMBP antisense RNA 1 (non-protein coding) (DNMBP-AS1), non-coding RNA [NR_024130]
ANGPTL4	5.696401	NM_139314	Homo sapiens angiopoietin-like 4 (ANGPTL4)
DUOX2	5.223868	NM_014080	Homo sapiens dual oxidase 2 (DUOX2)
IL1B	4.755472	NM_000576	Homo sapiens interleukin 1, beta (IL1B)
BLNK	4.511446	NM_013314	Homo sapiens B-cell linker (BLNK)
VTCN1	4.171324	NM_024626	Homo sapiens V-set domain containing T cell activation inhibitor 1 (VTCN1)
SFTPB	4.070699	NM_000542	Homo sapiens surfactant protein B (SFTPB)
SHE	4.028845	NM_001010846	Homo sapiens Src homology 2 domain containing E (SHE)
-	3.957717	DB335107	DB335107 SYNOV4 Homo sapiens cDNA clone SYNOV4009599 3', mRNA sequence [DB335107]
LOC100131581	3.913005	AK092544	Homo sapiens cDNA FLJ35225 fis, clone PROST2001116. [AK092544]
REPS2	3.894184	NM_004726	Homo sapiens RALBP1 associated Eps domain containing 2 (REPS2)
C7orf60	3.657602	NM_152556	Homo sapiens chromosome 7 open reading frame 60 (C7orf60)
DUOXA2	3.601023	NM_207581	Homo sapiens dual oxidase maturation factor 2 (DUOXA2)
C5orf4	3.594347	NM_032385	Homo sapiens chromosome 5 open reading frame 4 (C5orf4)
LOC283050	3.530351	NR_024431	Homo sapiens uncharacterized LOC283050 (LOC283050), transcript variant 1, non-coding RNA [NR_024431]
LOC100132354	3.422083	NR_024478	Homo sapiens uncharacterized LOC100132354 (LOC100132354), non-coding RNA [NR_024478]
ITM2B	3.385245	NM_021999	Homo sapiens integral membrane protein 2B (ITM2B)
ALDOC	3.37901	NM_005165	Homo sapiens aldolase C, fructose-bisphosphate (ALDOC)
C15orf48	3.370889	NM_032413	Homo sapiens chromosome 15 open reading frame 48 (C15orf48)

表2 FOXQ1敲低后, 下调倍数最大的20个基因

基因名称	倍性变化	mRNA编号	描述
TCF24	-7.699125	NM_001193502	Homo sapiens transcription factor 24 (TCF24)
MORN5	-6.097624	NM_198469	Homo sapiens MORN repeat containing 5 (MORN5)
GRK1	-5.3753	NM_002929	Homo sapiens G protein-coupled receptor kinase 1 (GRK1)
SPOCD1	-4.863297	NM_144569	Homo sapiens SPOC domain containing 1 (SPOCD1)
GDF6	-4.438107	NM_001001557	Homo sapiens growth differentiation factor 6 (GDF6)
SLC39A8	-4.372665	NM_001135147	Homo sapiens solute carrier family 39 (zinc transporter), member 8 (SLC39A8)
FAM48B2	-4.206	NM_001136233	Homo sapiens family with sequence similarity 48, member B2 (FAM48B2)
SERPINI1	-3.898295	NM_005025	Homo sapiens serpin peptidase inhibitor, clade I (neuroserpin), member 1 (SERPINI1)
MYBPH	-3.67658	NM_004997	Homo sapiens myosin binding protein H (MYBPH)
VPS53	-3.553322	NM_001128159	Homo sapiens vacuolar protein sorting 53 homolog (S. cerevisiae) (VPS53)
C3	-3.442721	NM_000064	Homo sapiens complement component 3 (C3)
-	-3.427454	-	HSU45880 X-linked inhibitor of apotosis protein {Homo sapiens} (exp = -1; wgp = 0; cg = 0), partial (29%) [THC2703908]
ZNF879	-3.426777	NM_001136116	Homo sapiens zinc finger protein 879 (ZNF879)
CCL20	-3.145884	NM_004591	Homo sapiens chemokine (C-C motif) ligand 20 (CCL20)
FMR1NB	-3.13995	NM_152578	Homo sapiens fragile X mental retardation 1 neighbor (FMR1NB)
GNN	-3.113418	NR_027249	Homo sapiens Grp94 neighboring nucleotidase pseudogene (GNN), non-coding RNA [NR_027249]
LOC400568	-3.076233	BC043554	Homo sapiens cDNA clone IMAGE:5176687. [BC043554]
DGCR14	-3.033284	NM_022719	Homo sapiens DiGeorge syndrome critical region gene 14 (DGCR14)
S100A8	-2.988219	NM_002964	Homo sapiens S100 calcium binding protein A8 (S100A8)
RN28S1	-2.977916	NR_003287	Homo sapiens RNA, 28S ribosomal 1 (RN28S1), ribosomal RNA [NR_003287]

表3 伴随FOXQ1敲低, 表达具有明显差异的基因pathway分析表

通路	基因	富集评分	伪发现率
上调通路
凋亡通路	CASP10/IL1B/IL1R1/IL3RA/PRKACB	2.5925	0.7182
细胞因子-细胞因子受体	ACVR2B/EGFR/IL1B/IL1R1/IL2RB/IL3RA/IL4/PDGFRB	2.0017	1.0000
FoxO通路	CCNG2/EGFR/FBXO32/S1PR4/SGK2	1.8017	1.0000
造血相关	IL1B/IL1R1/IL3RA/IL4	1.7900	1.0000
mTOR通路	DDIT4/RICTOR/TSC1	1.5428	1.0000
背腹轴形成相关	EGFR/PIWIL3	1.5423	1.0000
PI3K-Akt通路	DDIT4/EGFR/IL2RB/IL3RA/IL4/PDGFRB/SGK2/TSC1	1.4303	1.0000
甲状腺激素合成相关	DUOX2/DUOXA2/PRKACB	1.3422	1.0000
下调通路
细胞因子-细胞因子受体	CCL20/CXCL1/CXCL2/GDF6/IL17RA/IL8/LTB/TNF/TNFRSF11B/TPO	4.8088	0.0022
点头样受体通路	CARD9/CXCL1/CXCL2/IL8/TNF	4.3323	0.0044
基底细胞瘤相关	CTNNB1/GLI1/TP53/WNT16	3.2310	0.0275
核因子B通路	CXCL2/IL8/LTB/TNF	2.4180	0.1192
肿瘤坏死因子通路	CCL20/CXCL1/CXCL2/TNF	2.1262	0.2101
癌症转录失调相关	HIST1H3H/IL8/NR4A3/TP53/WNT16	2.0744	0.2152
趋化因子相关	CCL20/CXCL1/CXCL2/GRK1/IL8	1.9780	0.2463
补体、凝血相关	C3/PROC/SERPIND1	1.8955	0.2749
凋亡通路	PRKAR1B/TNF/TP53	1.6411	0.4281
肿瘤相关	CTNNB1/GLI1/IL8/TP53/TPM3/WNT16	1.5656	0.4776

表4 使用qRT-PCR验证芯片正确性的结果

相关性	Shcontrol	shFOXQ1
正相关
EGFR	1	1.313418
IL1B	1	1.81085
IL1R11	1	0.138536
Bag4	1	1.301339
SULT1C2	1	1.38831
DDIT4	1	9.849133
CCNG2	1	7.944722
负相关
FOXQ1	1	0.349492
CXCL1	1	0.232183
CXCL2	1	0.681442
PFAS1	1	5.388934
HRASLS5	1	0.622723
TNFRSF11B	1	0.946058
TNF	1	0.795536

¹与芯片结果不符.

表5 伴随FOXQ1下调, 上调倍数最大的10个microRNA

microRNA名称	倍比变化
hsa-miR-642b-3p	9.262590109
hsa-miR-550a-3-5p/ hsa-miR-550a-5p	9.084437272
hsa-miR-320d	6.849982996
hsa-miR-195-5p	6.109836399
hsa-miR-4524b-5p	5.078783166
hsa-miR-4506	4.961078779
hsa-miR-4419a	4.827465547
hsa-miR-3175	4.816025478
hsa-miR-4784	4.408422585
hsa-miR-548ap-5p/ hsa-miR-548j-5p	3.97387298

表6 伴随FOXQ1下调, 下调倍数最大的10个microRNA

microRNA名称	倍比变化
hsa-miR-5000-3p	-5.260356627
hsa-miR-206	-2.877979004
hsa-miR-145-3p	-2.773082367
hsa-miR-4796-5p	-2.745003957
hsv1-miR-H8-5p	-2.598690795
hsa-miR-19a-5p	-2.425592891
hsa-miR-302e	-2.311195074
hsa-let-7i-3p	-2.296028082
hsa-miR-491-3p	-2.265283812
hsa-miR-339-5p	-2.161523974