Carcinogenic Helicobacter pylori in gastric pre-cancer and cancer lesions: Association with tobacco-chewing

Table 1 Clinico-epidemiological details of subjects enrolled in study n (%)

Clinico-epidemiological characteristics	Normal	Pre-cancer	Cancer (adenocarcinoma)
Total subjects	61	30	65
Gender
Male	42 (68.9)	17 (56.7)	42 (64.6)
Female	19 (31.1)	13 (43.3)	23 (35.4)
Histopathological classification of lesions
Intestinal metaplasia	-	15 (50)	-
Dysplasia	-	15 (50)	-
Diffuse adenocarcinoma	-	-	44 (67.7)
Intestinal adenocarcinoma	-	-	21 (32.3)
Age (yr) (median)	7-85 (40)	13-78 (41.5)	22-90 (55)
Habits1
Tobacco chewers	28 (45.9)	15 (50)	24 (37)
Tobacco non-chewers	33 (54.1)	15 (50)	41 (63)
Tobacco smokers	22 (36.1)	11 (36.7)	20 (30.8)
Tobacco non-smokers	39 (64)	19 (63.3)	45 (69.2)
Alcoholic	15 (24.6)	8 (26.7)	16 (24.6)
Non-alcoholic	46 (75.4)	22 (73.3)	49 (75.4)
Multiple habits	22 (36.1)	9 (30)	16 (24.6)

¹Tobacco chewing habits include betel quid, areca nut, and/or pan masala use; multiple habits include two or more of any tobacco and/or alcohol habits.

Table 2 Primer sequences, primer annealing positions, expected lengths of amplified DNA products, and amplification cycle of polymerase chain reaction performed in the study

Primer	Sequence	Annealing position	Product length (bp)	Amplification cycle
Hp ureC-F	5’-AAG CTT TTA GGG GTG TTA GGG GTT T-3’	1293-1318	294	94 °C, 10’, 36 cycle (94 °C, 2’; 55 °C, 2’; 72 °C, 2’), 72 °C, 10’
Hp ureC-R	5’-AAG CTT ACT TTC TAA CAC TAA CGC-3’	1587-1563
Hp cagA-F	5’-AAT ACA CCA ACG CCT CCA AG-3’	2593-2612	400	94 °C, 4’, 39 cycle (94 °C, 2’; 59 °C, 2’; 72 °C, 2’), 72 °C, 10’
Hp cagA-R	5’-TTG TTG CCG CTT TTG CTC TC-3’	2992-2973
β-globin-F	5’-GAA GAG CCA AGG ACA GGT AC-3’	195-176	268	94 °C, 4’, 30 cycle (95 °C, 30’’; 55 °C, 30’’; 72 °C, 1’), 72 °C, 10’
β-globin-R	5’-CAA CTT CAT CCA CGT TCA ACC-3’	54-73

Table 3 Analysis of Helicobacter pylori gene expression in normal gastric tissues with clinico-epidemiological parameters n (%)

Clinico-epidemiological characteristics	Total cases, n	ureC positive	P	cagA positive	P	Hp +ve	P
Normal	61	33 (54.1)	-	21 (34.4)	-	37 (60.7)	-
Age (median 40 yr; range: 7-85 yr)
< 40	29	13 (44.8)	0.167	8 (27.6)	0.284	15 (51.7)	0.174
≥ 40	32	20 (62.5)		13 (40.6)		22 (68.8)
Gender
Male	42	18 (42.9)	0.009	14 (33.3)	0.789	22 (52.4)	0.049
Female	19	15 (78.9)		7 (36.8)		15 (78.9)
Habits1
Tobacco chewers	28	19 (67.9)	0.047	14 (50)	0.018	20 (71.4)	0.113
Tobacco non-chewers	33	14 (42.4)		7 (21.2)		17 (51.5)
Tobacco smokers	22	13 (59.1)	0.557	11 (50)	0.055	15 (68.2)	0.366
Tobacco non-smokers	39	20 (51.3)		10 (25.6)		22 (56.4)
Alcoholic	15	9 (60)	0.597	6 (40)	0.601	10 (66.7)	0.583
Non-alcoholic	46	24 (52.2)		15 (32.6)		27 (58.7)
Multiple habits	22	14 (63.6)	0.262	10 (45.5)	0.173	15 (68.2)	0.366

¹Tobacco chewing habits include betel quid, areca nut, and/or pan masala use; multiple habits include two or more of any tobacco and/or alcohol habit.

Table 4 Analysis of Helicobacter pylori gene expression in pre-cancer gastric tissues with clinico-epidemiological parameters n (%)

Clinico-epidemiological characteristics	Total cases, n	ureC positive	P	cagA positive	P	Hp +ve	P
Pre-cancer	30	27 (90)	0.0011	22 (73.3)	< 0.0011	28 (93.3)	< 0.0011
Age (median: 41.5 yr; range: 13-78 yr)
< 41.5	15	14 (93.3)	0.543	11 (73.3)	1.00	14 (93.3)	1.00
≥ 41.5	15	13 (86.7)		11 (73.3)		14 (93.3)
Gender
Male	17	15 (88.2)	0.713	14 (82.4)	0.201	16 (94.1)	0.844
Female	13	12 (92.3)		8 (61.5)		12 (92.3)
Types
Dysplasia	12	12 (100)	0.136	9 (75)	0.866	12 (100)	0.503
Intestinal metaplasia	18	15 (83.3)		13 (72.2)		16 (88.9)
Habits2
Tobacco chewers	15	13 (86.7)	0.543	12 (80)	0.409	14 (93.3)	1.000
Tobacco non-chewers	15	14 (93.3)		10 (66.7)		14 (93.3)
Tobacco smokers	11	11 (100)	0.165	10 (90.9)	0.098	11 (100)	0.265
Tobacco non-smokers	19	16 (84.2)		12 (63.2)		17 (89.5)
Alcoholic	8	8 (100)	0.271	7 (87.5)	0.290	8 (100)	0.377
Non-alcoholic	22	19 (86.4)		15 (68.2)		20 (90.9)
Multiple habits	9	9 (100)	0.232	8 (88.9)	0.207	9 (100)	0.338

¹Normal vs pre-cancer;

²Tobacco chewing habits include betel quid, areca nut, and/or pan masala use; multiple habits include two or more of any tobacco and/or alcohol habits.

Table 5 Analysis of Helicobacter pylori gene expression in adenocarcinoma gastric tissues with clinico-epidemiological parameters n (%)

Clinico-epidemiological Characteristics	Total cases, n	ureC positive	P	cagA positive	P	Hp +ve	P
Adenocarcinoma	65	33 (50.8)	< 0.0011	17 (26.2)	< 0.0011	34 (52.3)	< 0.0011
			0.7082		0.3122		0.3452
Age (median 55 yr; range: 22-90 yr)
< 55	31	14 (45.2)	0.388	10 (32.3)	0.285	15 (48.4)	0.546
≥ 55	34	19 (55.9)		7 (20.6)		19 (55.9)
Gender
Male	42	22 (52.4)	0.725	10 (23.8)	0.561	23 (54.8)	0.592
Female	23	11 (47.8)		7 (30.4)		11 (47.8)
Types
Diffuse	44	24 (54.5)	0.378	13 (29.5)	0.368	25 (56.8)	0.292
Intestinal	21	9 (42.9)		4 (19)		9 (42.9)
Habits3
Tobacco chewers	24	19 (79.2)	< 0.001	12 (50)	0.001	19 (79.2)	0.001
Tobacco non-chewers	41	14 (34.1)		5 (12.2)		15 (36.6)
Tobacco smokers	20	10 (50)	0.934	8 (40)	0.090	11 (55)	0.772
Tobacco non-smokers	45	23 (51.1)		9 (20)		23 (51.1)
Alcoholic	16	9 (56.2)	0.614	4 (25)	0.904	9 (56.3)	0.716
Non-alcoholic	49	24 (49)		13 (26.5)		25 (51)
Multiple habits	16	9 (56.2)	0.614	6 (37.5)	0.234	9 (56.3)	0.716

¹Pre-cancer vs adenocarcinoma;

²Normal vs adenocarcinoma;

³Tobacco chewing habits include betel quid, areca nut, and/or pan masala use; multiple habits include two or more of any tobacco and/or alcohol habits.