Impact of AKT1 polymorphism on DNA damage, BTG2 expression, and risk of colorectal cancer development

Zubair, Hina; Khan, Zahid; Imran, Muhammad

doi:10.2478/raon-2022-0031

Figures & Tables

Akt rs1130233 single nucleotide polymorphism in (A) Control and (B) colorectal cancer patients. Representative images have been shown. AA genotype (379 bp band); AG genotype (245 and 379 bp bands); GG genotype (245 bp band). The number above the lanes indicate subjects identity.C = Control, P = CRC Patient, M = DNA Marker

Figure 2

Association of AKT rs1130233 single nucleotide polymorphism with AKT expression and phosphorylation (A) Representative images of immunoblotting showing expression of AKT and pAKT in lymphocytes of GG, AG and AA carriers. (B) Mean densitometry profile of AKT and pAkt expression of different subjects of various genotypes. P < 0.05 GG vs AG, and P < 0.01 GG vs AA.

Figure 3

Association of AKT rs1130233 single nucleotide polymorphism BTG2 expression. Representative images have been shown. (A) BTG2 (360 bp) expression was determined in leukocytes of control individuals carrying GG, AG or AA genotypes of AKT1. GAPDH (158 bp) was used as a loading control. M = DNA Marker. (B) Mean densitometry profile of Btg2 mRNA expression of different subjects of various genotypes. P < 0.05 GG vs AG, and P < 0.01 GG vs AA.

Association of AKT1 rs1130233 single nucleotide polymorphism with DNA damage. (A) Representative images of comet assay have been shown. (B) Comet tail was quantified in leukocytes of control individuals carrying GG, AG or AA genotypes of AKT1. P < 0.05 GG vs AG, and P < 0.01 GG vs AA.

Frequencies of AKT1rs1130233 polymorphism and alleles in rectum cancer cases and control

Type of polymorphism	Genotype	n Rectum = 98 (%)	n Control = 197 (%)	P value	OR (95% CI)
	GG	30 (30.61)	90 (45.69)		Reference
Genotype frequency	AG	59 (60.20)	101 (51.27)	0.04	1.75 (1.04–2.96)
	AA	9 (9.19)	6 (3.04)	0.008	4.50 (1.48–13.69)
Dominant model	GG	30 (30.61)	90 (45.69)	0.01	1.91 (1.14–3.18)
	AG+AA	68 (69.39)	107 (54.31)
Recessive model	GG+AG	89 (90.81)	191 (96.96)	0.03	3.22 (1.11–9.32)
	AA	9 (9.19)	6 (3.04)

Primers sequences and amplification conditions for genes

Genes	Direction	Primer sequence	Amplification condition
BTG2	Forward	5^/-CCTGGGCAGAGAGTGAAAAG-3^/	95°C for 5 min, followed by 30
	Reverse	5^/-CCTTCCATCCTAACCCCAAT-3^/	cycles of 95°C for 30 s, 58°C for
	Forward	5^/-CCATGGAGAAGGCTGGGG-3^/	45 s, 72°C for 45 s and 72°C for
GAPDH	Reverse	5^/-CAAAGTTGTCATGGATGACC-3^/	10 min
	Forward	F1-5^/-ATAGGGAGTCATGGAGGGTTTG-3^/	95°C for 5 min, followed by 35
AKT1	Reverse	R1-5^/-CTTTACCAAATCCTGGTCACTGAA-3^/	cycles of 95°C for 30 s, 60°C for
	Forward	F2-5^/-AAAAAATTGATTGATGGGAGGAAG-3^/	45 s, 72°C for 45 s and 72°C for
	Reverse	R2-5^/-TAATCCCTGGCCTGCTCAG-3^/	10 min

Frequencies of AKT1 rs1130233 polymorphism and alleles in colon cancer cases and control

Type of polymorphism	Genotype	n = Colon 102 (%)	n Control = 197 (%)	P value	OR (95% CI)
	GG	30 (29.41)	90 (45.69)		Reference
Genotype frequency	AG	61 (59.80)	101 (51.27)	0.02	1.81 (1.08–3.05)
	AA	11 (10.79)	6 (3.04)	0.001	5.50 (1.87–16.15)
Dominant model	GG	30 (29.41)	90 (45.69)	0.006	2.02 (1.21–3.36)
	AG+AA	72 (70.59)	107 (54.31)
Recessive model	GG+AG	91 (89.21)	191 (96.96)	0.01	3.85 (1.38–10.73)
	AA	11 (10.79)	6 (3.04)

Demographic and clinical information of control subjects and colorectal cancer patients

Characteristics	Cases n = 200(%)	Control n = 197(%)
Age*
40 ≤	131 (65.5)	120 (60.91)
40 >	69 (34.5)	77 (39.09)
Sex*
Male	119 (59.50)	112 (57.50)
Female	81 (40.50)	85 (42.50)
Food consumption*
Mainly vegetables	106 (53.00)	97 (49.24)
Mixed Food	94 (47)	100 (50.76)
Smoking*
Ever	36 (18.00%)	29 (14.72%)
Never	164 (82.00%)	168 (85.28%)
Cancer family history*
Yes	27 (13.50)	17 (8.63)
No	173 (86.50)	180 (91.37)
Cancer Stages
I	1 (0.50)
II	33 (16.50)
III	112 (56.00)
IV	54 (27.00)

Gene and allele frequencies of AKT1 rs1130233 polymorphism and its association with colorectal cancer

Type of polymorphism	Genotype	n = Cases 200 (%)	n Control = 197 (%)	P value	OR (95% CI)
	GG	60 (30.00)	90 (45.69)		Reference
Genotype Frequency	AG	120 (60.00)	101 (51.27)	0.01	1.80 (1.18–2.74)
	AA	20 (10.00)	6 (3.04)	0.001	5.00 (1.90–13.18)
Dominant Model	GG	60 (30.00)	90 (45.69)
	AG+AA	140 (70.00)	107 (54.31)	0.001	1.96 (1.30–2.96)
Recessive Model	GG+AG	180 (90.00)	191 (96.96)
	AA	20 (10.00)	6 (3.04)	0.01	0.28 (0.11–0.72)
Allele Frequency	G A	0.6000 0.4000	0.7132 0.2868

Impact of AKT1 polymorphism on DNA damage, BTG2 expression, and risk of colorectal cancer development

Figures & Tables

Figure 1