Cancer Genetics and Cytogenetics 196 (2010) 201e203
Letter to the editor
Incidence of the L858R and G719S mutations
of the epidermal growth factor receptor oncogene
in an Ecuadorian population with lung cancer
The World Health Organization indicates that lung
cancer (LC) represents the main cause of death in many
developed countries. In Ecuador, according to the National
Cancer Registry, LC is the fourth cause of death among
malignant tumors, followed by gastric, pancreatic, and
breast cancer [1]. Moreover, three provinces present the
highest percentages of the disease d Guayas (32.8%), Pi-
chincha (20.7%), and Manabı́ (7.5%) [1]. It is estimated
that this tendency will increase in the next years, and would
place this disease as the principal cause of death by
neoplasia. Therefore, it is important to perform the first
mutation research in relation to the tyrosine kinase domain
(TKD) of the EGFR gene in Ecuador.
The ERBB family is one of the receptor systems located
in the cytoplasmatic membrane that presents intrinsic tyro-
sine kinase activity (TKA). The epidermal growth factor
receptor (EGFR) is located in 7p12.1wp12.3 and encodes
a precursor protein of 1,210 amino acids [2]. This integral
membrane glycoprotein of 170 kDa is formed by the extra-
cellular membrane receptor domain, the lipophilic trans-
membrane domain, and the intracellular domain [2]. The
phosphorylation of the cytosolic TKD initializes the down-
stream of intracellular signal transduction that regulates cell
proliferation, cell differentiation, and angiogenesis [3]. The
somatic mutations of the EGFR gene are grouped in the
ATP kinase domain. These precise mutations constitute
one of the most common genetic anomalies found in this
gene, due to the fact that these variants correlate themselves
with the presence of oncogenic factors and apoptotic
mechanisms [4].
The population studied consisted of 189 male and
female Ecuadorians. A total of 80 tumor samples were ob-
tained through the Department of Pathology at Hospital
Carlos Andrade Marin, and the control group included
109 healthy volunteers who were selected randomly.
Genotype was determined through the polymerase chain
reaction restriction fragment length polymorphism tech-
nique. The SacI restriction enzyme (New England BioLabs,
Ipswich, MA) was used to identify the presence of G719S,
which consists of the exchange of glycine (G) for serine (S)
in the 719 codon, while the Sau96I enzyme allows for the
detection of L858R, which consists of the exchange of
leucine (L) for arginine (R) in the 858 codon.
0165-4608/10/$ e see front matter Ó 2010 Elsevier Inc. All rights reserved.
doi:10.1016/j.cancergencyto.2009.10.007
The bio-pathologic parameters considered in this
research were age, sex, smoking, and histotype. The
average age of the individuals with LC at the time of diag-
nosis was 68 years, which proved that old age is a risk
factor for acquiring this disease. With regard to gender,
men were 35% more affected than women, but 54% of
women showed EGFR mutations versus 46% of men. As
far as smoking was concerned, 69% of the individuals diag-
nosed with LC used to smoke. With regard to the presence
of EGFR mutations, 15 of the individuals used to smoke
and 23 did not. Finally, according to classification by histo-
type, 94% of the affected individuals had nonesmall cell
lung cancer (NSCLC), and the most representative histo-
type was adenocarcinoma with 91%, of which 49% of indi-
viduals with adenocarcinoma had EGFR mutations. On the
other hand, 6% of the affected individuals had small cell
lung cancer (SCLC), and none of them had EGFR
mutations.
Using the odds ratio (OR) statistitical analysis, it was
determined that LC risk for smokers is 117.7 times higher
for smokers than for nonsmokers. In other words, it is
evident that smoking is the principal cause of LC diagnoses
[5]. Nevertheless, we were able to observe that 61% of indi-
viduals with EGFR mutations were not active smokers and
had an OR Z 0.033, which means that there was no relative
risk between smoking and the presence of variants in the
TKD [6,7].
The studied individuals with L858R and G719S showed
the adenocarcinoma histotype, representing the first study
in which all of the individuals with LC had this histotype.
Some international research studies, such as the one done
by Sharma et al. [8], state that even though most of the
affected individuals with EGFR mutations occur in
NSCLC, those mutations are also found in SCLC, despite
their low incidence [8]. Regarding the L858R mutation,
we could observe a higher frequency of the leucine allele
in controls than in the affected individuals. In fact, the argi-
nine allele frequency was 0.34 in individuals with LC and
0.09 in healthy ones. The chi square test determined the
existence of a highly significant difference related to the
presence of L858R variant between affected and healthy
individuals (c2 Z 22.4; P ! 0.001). Therefore, the pres-
ence of this variant is associated with the development of
202 Letter to the editor / Cancer Genetics and Cytogenetics 196 (2010) 201e203

Table 1 Table 3
Genotype distribution and allele frequency of L858R variant Statistical analysis: chi square and odds ratio tests
Group Genotype Individuals Percent Genotypic Allelic L858R L/L L/R R/R Chi square OR
frequency frequency Affected 63% 6% 31% 22.4 a
5.9 95%
Affected L/L 50 63 0.63 0.66 CI Z 2.7e13.1
(n Z 80) L/R 5 6 0.06 Control 91% 1% 8% P ! 0.001 P ! 0.001
R/R 25 31 0.31 0.34 G719S G/G G/S S/S c2 OR
Control L/L 99 91 0.91 0.91 Affected 91% 3% 6% 2.2b 2.5 95%
(n Z 109) L/R 1 1 0.01 CI Z 0.71e8.9
R/R 9 8 0.08 0.09 Control 96% 1% 3% P O 0.05 P O 0.05
a
Significant.
b
Not Significant.
LC. The OR test establishes that it is 5.9 times more likely
to have LC when a mutated genotype is present in both allele frequency in the patients was 0.66 and in the controls
heterozygous carriers L/R and the homozygous ones R/R. was 0.91, whereas the arginine allele frequency in the
On the other hand, with regard to the G719S variant, the patients was 0.34 and in the controls was 0.09.
glycine allele frequency was mainly observed in control Table 2 lists the genotypic distribution and allele frequen-
individuals more than in affected individuals; meanwhile, cies of the G719S variants. A total of 105 patients were
the serine frequency was 0.08 for cases and 0.04 for homozygous for the glycine allele (G/G), 1 was heterozy-
controls. The chi square test determined that there was no gous (G/S), and 3 were homozygous for the serine allele
significant difference related to the presence of these muta- (S/S). As for the affected individuals, 73 were homozygous
tions between cases and controls (c2 Z 2.2; P O 0.05). for the glycine allele (G/G), 2 were heterozygous (G/S),
Therefore, the presence of the G719S mutation is not asso- and 5 were homozygous for the serine allele (S/S). The
ciated with the development of LC in this population. The glycine allele frequency in the patients was 0.92 and in the
connection between the G/S and S/S genotypes, however, is controls was 0.96, whereas the serine allele frequency in
evident in the risk of developing LC [9]. the patients was 0.08 and in the controls was 0.04.
In the Ecuadorian population, L858R and G719S muta- Table 3 shows the statistical analysis using the chi
tions are mainly found in NSCLC, in women, nonsmokers, square test to determine the level of significance of
and individuals with adenocarcinoma. At present, researches L858R and G719S variants between affected and healthy
are conducting studies on infections with human papiloma individuals with a significance level of 5%, and OR tests
virus (HPV), nutritional status, genetic susceptibility, and were calculated with 95% confidence intervals (95%CI).
immunologic infections to determine whether any of these With concern to the L858R variant, subsequent to corre-
is a contributing factor in nonsmoking women who have lating the data of the 80 individuals with LC and the 109
LC and EGFR mutations [6]. Concerning EGFR mutations healthy individuals, a value of c2 Z 22.4 (P ! 0.001)
in Ecuadorians with LC, a high impact of L858R variant was obtained. With the G719S variant, a value of
was observed, unlike the G719S mutation. This indicates that c2 Z 2.2 (P O 0.05) were obtained. The presence of the
the TK mutations significantly influence individuals L858R and G719S mutations were correlated with the
diagnosed with this neoplasia [8]. OR obtained. Out of a total of 80 individuals with LC, 30
Table 1 lists genotype distribution and allele frequencies of them showed a mutant allele (L/R or R/R), and the re-
of the L858R variant by caseecontrol status. A total of 99 maining 50 had a normal allele (L/L). In contrast, out of
healthy individuals were homozygous for the leucine allele a total of 109 healthy individuals, 10 had a mutant allele,
(L/L), 1 was heterozygous (L/R), and 9 were homozygous and 99 of them showed a normal allele. The statistical test
for the arginine allele (R/R). On the other hand, among the was applied, obtaining an OR of 5.9 (OR Z 5.9, 95%CI
individuals with LC, 50 were homozygous for the leucine 2.7e13.1, P ! 0.001). Regarding the G719S variant, out
allele (L/L), 5 were heterozygous (L/R), and 25 were of 109 control individuals, 4 had mutant allele (G/S or S/
homozygous for the arginine allele (R/R). The leucine S) and 105 did not (G/G). Moreover, out of 80 affected
individuals, 7 showed a mutant allele and 73 had a normal
allele, obtaining an OR of 2.5 (OR Z 2.5 95%CI 0.71e8.9,
Table 2
Genotype distribution and allele frequency of G719S variant P O 0.05). In addition, the OR test was also applied to
determine the relative risk of affected individuals with the
Group Genotype Individuals Percent Genotypic Allelic
frequency frequency presence of the G719S and L858R variants related to
smoking. Out of 80 individuals with LC, 15 had been
Affected G/G 73 91 0.91 0.92
(n Z 80) G/S 2 3 0.03 exposed to cigarette smoking and had mutant alleles. A
S/S 5 6 0.06 0.08 total of 23 showed mutant alleles and had no exposure to
Control G/G 105 96 0.96 0.96 cigarette smoking. In addition, 40 of them did not have
(n Z 109) G/S 1 1 0.01 mutant alleles (L/L, G/G), but had exposure to cigarette
S/S 3 3 0.03 0.04
smoking. Two of them did not show mutant alleles and
 Letter to the editor / Cancer Genetics and Cytogenetics 196 (2010) 201e203 203

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