Expression of miR-1304 in patients with esophageal carcinoma and risk factors for recurrence

Table 1 Comparison of baseline data

Factors	Normal group (n = 50)	Patient group (n = 78)	t / χ2	P value
Gender			0.161	0.688
Male	30 (60.00)	44 (56.41)
Female	20 (40.00)	34 (43.59)
Age (yr)	60.4 ± 4.8	58.7 ± 5.4	0.310	0.757
Smoking history			0.295	0.587
Yes	30 (60.00)	43 (55.13)
No	20 (40.00)	35 (44.87)
History of alcoholism			0.081	0.776
Yes	8 (16.00)	14 (17.95)
No	42 (84.00)	64 (82.05)
Location
The upper thoracic area		19 (24.36)
The middle thoracic area		34 (43.59)
The lower thoracic area		25 (32.05)
Tumor size
≥ 3 cm		45 (57.69)
< 3 cm		33 (42.31)
Differentiation
Low differentiation		25 (32.05)
Moderate + high differentiation		53 (67.95)
TNM stage
Stage I		27 (34.62)
Stage II + III		51 (65.38)

Table 2 Relationship between miR-1304 and pathological data

Factors	High expression (n = 39)	Low expression (n = 39)	χ2	P value
Gender			0.834	0.361
Male (n = 44)	20 (51.28)	24 (61.54)
Female (n = 34)	19 (48.72)	15 (38.46)
Age (yr)			0.821	0.365
≥ 60 (n = 38)	17 (43.59)	21 (53.85)
< 60 (n = 40)	22 (56.41)	18 (46.15)
Lesion location			1.523	0.467
The upper thoracic area (n = 19)	10 (25.64)	9 (23.08)
The middle thoracic area (n = 34)	19 (48.72)	15 (38.46)
The lower thoracic area (n = 25)	10 (25.64)	15 (38.46)
Tumor size			6.356	0.001
≥ 3 cm (n = 45)	28 (71.79)	17 (43.59)
< 3 cm (n = 33)	11 (28.21)	22 (56.41)
Differentiation			4.768	0.029
Low differentiation (n = 25)	17 (43.59)	8 (20.51)
Moderate + high differentiation (n = 53)	22 (56.41)	31 (79.49)
TNM stage			6.854	0.009
Stage I (n = 27)	8 (20.51)	19 (48.72)
Stage II + III (n = 51)	31 (79.49)	20 (51.28)

Table 3 Receiver operating characteristic parameters

Parameters	AUC	95%CI	P value	Sensitivity (%)	Specificity (%)	Cut-off
Tumor size	0.710	0.591-0.829	0.002	42.42	95.56	< 1.158
Differentiation	0.773	0.665-0.881	0.001	77.36	68.00	< 1.363
TNM staging	0.788	0.687-0.889	< 0.001	47.06	96.30	> 1.393

AUC: Area under the curve.

Table 4 Univariate analysis

Factors	Recurrence group (n = 33)	Non-recurrence group (n = 45)	t /χ2	P value
Gender			2.792	0.095
Male (n = 44)	15 (45.45)	29 (64.44)
Female (n = 34)	18 (54.55)	16 (35.56)
Age (yr)			0.179	0.672
≥ 60 (n = 38)	17 (51.52)	21 (46.67)
< 60 (n = 40)	16 (48.48)	24 (53.33)
Smoking history			0.694	0.405
Yes (n=43)	20 (60.61)	23 (51.11)
No (n=35)	13 (39.39)	22 (48.89)
History of alcoholism			1.319	0.251
Yes (n=14)	4 (12.12)	10 (22.22)
No (n=64)	29 (87.88)	35 (77.78)
Lesion location			0.602	0.740
The upper thoracic area (n = 19)	7 (21.21)	12 (26.67)
The middle thoracic area (n = 34)	16 (48.49)	18 (40.00)
The lower thoracic area (n = 25)	10 (30.30)	15 (33.33)
Tumor size			7.648	0.006
≥ 3 cm (n = 45)	25 (75.76)	20 (44.44)
< 3 cm (n = 33)	8 (24.24)	25 (55.56)
Differentiation			7.093	0.008
Low differentiation (n = 25)	16 (48.48)	9 (20.00)
Moderate + high differentiation (n = 53)	17 (51.52)	36 (80.00)
TNM staging			6.825	0.009
Stage I (n = 27)	6 (18.18)	21 (46.67)
Stage II + III (n = 51)	27 (81.82)	24 (53.33)
Adjuvant therapy			0.946	0.331
Chemotherapy (n = 52)	24 (72.73)	28 (62.22)
Radiotherapy (n = 26)	9 (27.27)	17 (37.78)
Expression of miR-1304			8.877	0.003
High expression (n = 39)	23 (69.70)	16 (35.56)
Low expression (n = 39)	10 (30.30)	29 (64.44)

Table 5 Assignment

Factors	Assignment
Tumor size	≥ 3 cm = 1, < 3 cm = 0
Differentiation	Low differentiation = 1, Moderate + high differentiation = 0
TNM stage	Stage I = 1, stage II + III = 0
Expression of miR-1304	High differentiation = 1, low differentiation = 0
Recurrence	Recurrence = 1, non-recurrence = 0

Table 6 Logistic regression analysis

Factors	β	SD	χ2	P value	Exp (HR)	HR 95%CI
						Up	Down
Tumor size	1.861	0.646	8.297	0.004	6.429	1.812	22.803
Differentiation	1.811	0.665	7.414	0.006	6.114	1.661	22.506
TNM stage	-1.801	0.675	7.118	0.008	0.165	0.044	0.620
Expression of miR-1304	1.838	0.627	8.586	0.003	6.286	1.838	21.501

CI: Credibility interval.

Table 7 Cox regression analysis

Factors	3-yr univariate Cox		3-yr multivariate Cox
	P value	H (95%CI)	P value	HR (95%CI)
Gender (male vs female)	0.803	1.075 (0.608-1.903)
Age (≥ 60 yr vs < 65 yr)	0.534	0.835 (0.473-1.474)
Lesion location (upper vs middle vs lower)	0.427	1.166 (0.798-1.705)
Tumor size (≥ 3 cm vs < 3 cm)	0.000	3.473 (1.827-6.601)	0.010	2.402 (1.237-4.665)
Differentiation (low differentiation vs moderate + high differentiation)	0.000	3.09 (1.724-5.539)	0.015	2.153 (1.159-4.002)
TNM stage (stage I vs stage II + III)	0.060	0.543 (0.287-1.027)
Adjuvant therapy plan (chemotherapy vs radiotherapy)	0.939	1.025 (0.549-1.911)
Recurrence (recurrence vs non-recurrence)	0.002	2.561 (1.408-4.657)	0.034	1.949 (1.05-3.619)
miR-1304 (high expression vs low expression)	0.001	2.614 (1.448-4.717)	0.036	1.93 (1.044-3.565)

Table 8 Target genes of miR-1304

Gene	Target gene
miR-1304	LYPD3, MYC, LMNB1, KLHL15, ZNF99, SPRYD4, CCNT2, PLEKHF2, DSEL, ADIPOR2, CBX5, DDX3X, SMAD5, DARS, PCGF3, FAM83H, RRAS, PDE3A, CALM2, and TMBIM6

Table 9 Gene ontology enrichment analysis

ID	Term	Gene	Count	P value
GO: 0043392	Negative regulation of DNA binding	WFIKKN2, SP100, HAND2, HMOX1, HMGA2	5	0.001
GO: 0051091	Positive regulation of sequence-specific DNA binding transcription factor activity	IL6, SP100, HIPK2, PYCARD, HMGA2, TRIM22, TCF3, ATF2	8	0.001
GO: 0000122	Negative regulation of transcription from RNA polymerase II promoter	ATF7IP, SATB1, BACH2, SP100, GABPA, DICER1, YBX3, KLF16, ZEB2, HMGA2, ZNF345, ATF2, SUV39H2, VDR, OLIG3, MLX, PRMT6, HIPK2, ZNF431, TCF3, SMARCA2, SUDS3	22	0.002
GO: 0006351	Transcription, DNA-templated	XRCC5, ZNF555, IRX5, CCNT1, YBX3, BBX, TTLL5, ZNF250, ZEB2, ZNF652, ZNF184, VDR, OLIG3, PRMT6, ZNF426, KDM3B, BAZ2A, TCF3, MAP2K6, ATF7IP, SATB1, SP100, ZNF620, ZNF92, KLF16, SCAI, TRIM22, ATMIN, ZNF585B, FAM208A, SUV39H2, ZBTB25, EYA4, ZNF439, MLX, PARP14, HIPK2, ZNF431, ZNF432, SMARCA2, FOXD4L6, SUDS3, ZNF573, ZBTB8A	44	0.002
GO: 0006355	Regulation of transcription, DNA-templated	ZNF555, IRX5, YBX3, BBX, ZNF250, ZNF652, ATF2, VDR, ZNF184, ZNF426, KDM3B, BAZ2A, INSR, TCF3, MAP2K6, SATB1, ZNF620, ZNF92, SCAI, AFF1, HMGA2, TRIM22, ZNF585B, FAM208A, ZBTB25, EYA4, TULP4, ZNF439, PARP14, CDKN2AIP, MLX, ZNF431, ZNF432, SMARCA2, ZBTB8A, ZNF573	36	0.003
GO: 0005138	Interleukin-6 receptor binding	IL6, PYCARD, ADAM17	3	0.004
GO: 0015031	Protein transport	SLC7A6OS, RAB9A, RAB3D, KIF17, DUOXA1, RAB39A, VPS52, PKDCC, HOOK3, GOPC, SNX30, SNX20, EXOC6B	13	0.011
GO: 0050892	Intestinal absorption	F11R, VDR, KCNQ1	3	0.016
GO: 0042147	Retrograde transport, endosome to Golgi	SPAG9, RAB9A, STX16, VPS52, VPS26A	5	0.017
GO: 0032259	Methylation	METTL8, TRMT10B, SETD9, CARNMT1, NNMT	5	0.020
GO: 0032290	Peripheral nervous system myelin formation	NCMAP, DICER1	2	0.028
GO: 0005509	Calcium ion binding	PCDH11Y, PCDH11X, RPH3AL, DAG1, STIM1, MMP17, FSTL4, TTN, IQGAP1, CALU, ITPR2, PLSCR1, CALML4, TPT1, SYTL2, EGF, CALM2, CACNA1B	18	0.029
GO: 0008544	Epidermis development	SATB1, STS, ELF3, INSR, SCEL	5	0.033
GO: 0003677	DNA binding	XRCC5, ZNF555, AGFG1, KIAA1958, CCNT1, YBX3, BBX, ZNF250, ZEB2, ZNF345, ZNF652, ZNF184, VDR, OLIG3, HIST1H4C, ZNF426, BAZ2A, TCF3, SATB1, SP100, ZNF620, CHTF8, GABPA, HMGA2, ZBTB25, PLSCR1, ZNF439, RFC2, MLX, HIPK2, RAD18, ZNF432, ZNF573, ZBTB8A	34	0.035
GO: 0046872	Metal ion binding	ZNF555, MGAT5B, AGFG1, DICER1, ZNF250, ZEB2, ZNF345, ZNF652, ATF2, ZFC3H1, ZNF184, CLEC17A, FGG, HMOX1, ZC3H12B, ZNF426, KDM3B, STS, KCND3, NRXN3, ZNF620, ZNF92, KLF16, RPH3AL, ATMIN, ZNF585B, ZBTB25, EYA4, ZNF439, GNAQ, ADAM17, ZNF431, YME1L1, ZNF432, ADAM12, ZNHIT6, CACNA1D, UGP2, ZBTB8A, ZNF573	40	0.042
GO: 0070644	Vitamin D response element binding	VDR, TCF3	2	0.042
GO: 0005794	Golgi apparatus	STS, RAB39A, FGFRL1, VPS52, PKDCC, PRKG1, SART1, CALU, HOOK3, PLSCR1, NRAS, COPB1, STX16, GOPC, CNTNAP2, CAND1, B4GALT7, TRAPPC3, SLC30A7	19	0.044
GO: 0007173	Epidermal growth factor receptor signaling pathway	NRAS, ADAM17, EGF, IQGAP1	4	0.045

Table 10 Kyoto Encyclopedia of Genes, and Genomes pathway enrichment analysis

ID	Term	Genes	Count	P value
hsa04912	GnRH signaling pathway	NRAS, GNAQ, CACNA1D, MAP2K6, CALM2, ITPR2	6	0.007
hsa04725	Cholinergic synapse	NRAS, GNAQ, CACNA1D, KCNQ1, ITPR2, CACNA1B	6	0.015
hsa04022	cGMP-PKG signaling pathway	GNAQ, PRKG1, CACNA1D, INSR, CALM2, ITPR2, ATF2	7	0.017
hsa04925	Aldosterone synthesis and secretion	GNAQ, CACNA1D, CALM2, ITPR2, ATF2	5	0.021
hsa04728	Dopaminergic synapse	GNAQ, CACNA1D, CALM2, ITPR2, ATF2, CACNA1B	6	0.026
hsa05164	Influenza A	IL6, AGFG1, PYCARD, DNAJC3, MAP2K6, HLA-DRA, ATF2	7	0.026
hsa04540	Gap junction	NRAS, GNAQ, PRKG1, EGF, ITPR2	5	0.028
hsa04066	HIF-1 signaling pathway	IL6, PFKFB3, HMOX1, EGF, INSR	5	0.037
hsa04915	Estrogen signaling pathway	NRAS, GNAQ, CALM2, ITPR2, ATF2	5	0.040
hsa04922	Glucagon signaling pathway	GNAQ, CALM2, G6PC2, ITPR2, ATF2	5	0.040
hsa04730	Long-term depression	NRAS, GNAQ, PRKG1, ITPR2	4	0.043