Association between the polymorphisms of angiotensin converting enzyme (Peptidyl-Dipeptidase A) INDEL mutation (I/D) and Angiotensin II type I receptor (A1166C) and breast cancer among post menopausal Egyptian females

Peer review under responsibility of Alexandria University Faculty of Medicine.

Available online 7 January 2014

Figures & data

Figure 1 Agarose gel electrophoresis showing (I/D) polymorphisms of the ACE gene. (D) and (I) alleles were identified by the presence of 190-bp and 490-bp fragments, respectively. Lane A: 100–3000 DNA ladder, Lane B: (DD), Lane C: (ID), Lane D: (II), Lane E: (DD), Lane F: (II), Lane G: (ID), Lane H: (DD), Lane I: (ID).

Figure 2 Allele specific amplification using insertion-specific sequences. The (I) allele produced a 335 bp amplicon. Lane A (upper and lower raws): 100–3000 bp DNA ladder. Lane B (upper raw): II (positive control). Lane B (lower raw): ID mistyped as DD. Lanes C–H (upper and lower raws): correctly typed DD.

Figure 3 PCR product on 2% agarose gel at 850 bp with (100–3000 bp) DNA ladder marker.

Figure 4 Dde I restriction digestion of the PCR product Homozygote (CC) produced three bands (600, 140 and 110 bp long) homozygote (AA) produced two bands (600 and 250 bp long). Heterozygote (AC) produced all four bands. Lane A: 100–3000 bp DNA ladder. Lanes B–D, F, G: homozygous (AA). Lane E: homozygous (CC). Lanes H and I: heterozygous (AC).

Table 1 Comparison of AT₁R A1166C SNP genotype frequencies among the studied groups.

A1166C AT1R SNP genotype	Control group (n = 50)	Patients group (n = 70)	Exact p-value
AA (n = 75; 62.5%)	38 (76%)	37 (52.9%)	p = 0.021108 ∗
AC (n = 43; 35.8%)	12 (24%)	31 (44.3%)
CC (n = 2; 1.7%)	0 (0%)	2 (2.9%)
Total (n = 120; 100%)	50 (100%)	70 (100%)	120

Table 2 Comparison of AT₁R A1166C SNP allele frequencies among the studied groups.

A1166C alleles	Control group (n = 50)	Patients group (n = 70)	Total	Exact p-value	Odds ratio (95% CI)
Allele A	88 (88%)	105 (75%)	193 (80.4%)	p = 0.0134 ∗	2.4444 (1.1967–4.9931)
Allele C	12 (12%)	35 (25%)	47 (19.6%)
Total	100 (100%)	140 (100%)	240 (100%)

Table 3 Assessment of the risk of developing breast cancer according to AT₁R + A1166C SNP genotypes in different models of inheritance.

Assumed model of inheritance	Model (unexposed vs exposed)	Genotype	Control group n = 50 (100%)	Patients group n = 70 (100%)	Exact p-value	Odd’s ratio (95% CI)
^aRecessive	(AA and AC) vs CC	AA and AC	50 (100%)	68 (97.1%)	–	–
		CC	0.0 (0.00%)	2 (2.9%)
^bDominant	AA vs (AC and CC)	AA	38 (76%)	37 (52.9%)	0.013^∗	2.8243 (1.2679–6.2913)
		AC and CC	12 (24%)	33 (47.1%)

Table 4 Variables included in the final logistic regression model with breast cancer risk as the dependent variable.

Variable	Wald	p-Value	Exp (B)	95% CI for Exp (B)
Body mass index (BMI)	11.146	0.001	1.265	(1.102–1.452)
A1166C AT1R SNP genotype (AC and CC versus AA)	6.875	0.009	3.419	(1.364–8.569)

Table 5 Comparison of ACE I/D genotype frequencies among the studied groups.

ACE I/D genotype	Control group (n = 50)	Patients group (n = 70)	Exact P-value
II (n = 21; 17.5%)	8 (16%)	13 (18.6%)	p = 0.940
ID (n = 49; 40.8%)	21 (42%)	28 (40%)
DD (n = 50; 41.7)	21 (42%)	29 (41.4%)
Total (n = 120; 100%)	50 (100%)	70 (100%)	120

Table 6 Comparison of ACE I/D allele frequencies among the studied groups.

ACE I/D	Control group (n = 50)	Patients group (n = 70)	Total	Exact P-value	Odds ratio (95% CI)
Insertion (I)	37 (37%)	54 (39%)	91 (38%)	0.893	0.9353 (0.5507–1.5886)
Deletion (D)	63 (63%)	86 (61%)	149 (62%)
Total	100 (100%)	140 (100%)	240 (100%)