Helicobacter pylori-induced miR-136 is a potential predictor of early-stage gastric cancer

Table 1 Characteristics of the patients with gastric cancer and gastritis

Characteristics	Gastric cancer (n = 60)	Gastritis (n = 90)			P value
		NAG	CAG	PLGC
Age, years					0.097
18-45	16 (27)	16 (18)	11 (12)	12 (13)
> 45	44 (73)	14 (16)	19 (21)	18 (20)
Sex					0.244
Male	42 (70)	16 (18)	21 (23)	23 (25)
Female	18 (30)	14 (16)	9 (10)	7 (8)
H. pylori					< 0.001
Negative	30 (50)	30 (33)	0 (0)	0 (0)
Positive	30 (50)	0 (0)	30 (33)	30 (33)
Smoking					0.965
Yes	34 (57)	18 (20)	16 (18)	17 (19)
No	26 (43)	12 (13)	14 (16)	13 (14)
Drinking					0.881
Yes	37 (62)	19 (22)	17 (19)	20 (22)
No	23 (38)	11 (12)	13 (14)	10 (11)
Family history					0.569
Yes	36 (60)	14 (16)	17 (19)	19 (21)
No	24 (40)	16 (18)	13 (14)	11 (12)
OLGA					< 0.001
I	0 (0)	23 (26)	17 (19)	1 (1)
II	1 (2)	6 (7)	11 (12)	5 (6)
III	27 (45)	1 (1)	2 (2)	13 (14)
IV	32 (53)	0 (0)	0 (0)	11 (12)
OLGIM					< 0.001
I	0 (0)	17 (19)	11 (12)	0 (0)
II	2 (3)	12 (13)	15 (17)	3 (3)
III	28 (47)	1 (1)	4 (4)	11 (13)
IV	30 (50)	0 (0)	0 (0)	16 (18)

NAG: Non-active gastritis without Helicobacter pylori infection; CAG: Chronic gastritis with Helicobacter pylori-infection; PLGC: Precursor lesions of gastric cancer with Helicobacter pylori-infection; H. pylori: Helicobacter pylori; OLGA: Operationally Defined Staging Criteria for Gastritis; OLGIM: Operative Link for Gastritis Intestinal Metaplasia.

Table 2 Sequences of primers and microRNAs

Gene		Primer sequence (5’-3’)
miR-136	Forward	ACACTCCAGCTGGGACTCCATTTGTTTTG
	Reverse	CTCAACTGGTGTCGTGGAGTCGGCAATCAGTTGAGTCCATCAT
U6	Forward	CTCGCTTCGGCAGCACA
	Reverse	AACGCTTCACGAATTTGCGT
PDCD11	Forward	GAAGCTACCATGGTTGCAACAGTACAGG
	Reverse	CCTGTACTGTGAATTCGATGGTAGCTTCC
GAPDH	Forward	GAGGGATTTTTGTTTTTATGTTTTG
	Reverse	TATATACACCAACAACTCTCTCCAA
miR-136 mimics	Forward	ACUCCAUUUGUUUUGAUGAUGGA
	Reverse	UCCAUCAUCAAAACAAAUGGAGU
miR-NC	Forward	UUUGUACUACACAAAAGUACUG
	Reverse	CAGUACUUUUGUGUAGUACAAA
miR-136 inhibitor		UCCAUCAUCAAAACAAAUGGAGU
NF-κB WT		GGGGCAACGGCAGGGGAATTCCCCTCTCCTTA
NF-κB Mut		GGGGCAACGGCAGATCTATTCCCCTCTCCTTA

NF-κB: Nuclear factor kappa-B; WT: Wild-type; Mut: Mutated binding site; NC: Control.