VOLUME - 7

CHIEF EDITOR

Sergiy Fedorov

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 RESEARCH TRENDS
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MED CAL SCIENCES
Volume - 7

Chief Editor
Sergiy Fedorov (MD, Ph.D., MBA, D.Sc.)
Professor of Therapy and Family Medicine Department of Postgraduate
Faculty, Ivano-Frankivsk National Medical University, Ukraine

AkiNik Publications
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 Contents

Chapters Page No.

1. Genetic Variation and Cardiovascular Adaptation to Exercise 01-21
(Parul Khare, Manpreet Kaur Taluja and Prabhat K. Budholia)

2. Management of Ulcerative Colitis: Present and Future

Treatments 23-38
(Arpan Kumar Maiti, Spoorthi B.C., Shashwati Ghosh and Ishita Saha)

3. Graphene Quantum Dots for Application in Biosensing and

Bioimaging 39-53
(Narendra B. Patil, Ketan B. Patil and Paresh A. Patil)

4. Hypertension and Management 55-85

(Dr. Naga Subrahmanyam S, Dr. Tagoore Vijaya Lakshmi D, Dr. G.V.
Nagaraju, K.S.S.S. Swarna Suma and G. Jigeesha Lakshmi)

5. Rutin: A Type of Flavonoid with Immense Health Benefits 87-108

(Rajesh Prasad and Surya Bali Prasad)

6. Lung Cancer and Tobacco Smoking: Changes in Epigenetic

DNA Methylation 109-126
(Diana Monserrat Aguilar-Beltrán, Brenda Ugalde-Villanueva, Alma Delia
Bertadillo-Jilote, David Gustavo García-Gutiérrez, Marco Antonio Meraz-Ríos
and Karla Isabel Lira-De León)

7. Nephrotic Syndrome 127-146

(Rimpi Devi)
 Chapter - 6
Lung Cancer and Tobacco Smoking: Changes in
Epigenetic DNA Methylation

Authors
Diana Monserrat Aguilar-Beltrán
Maestría En Química Clínica Diagnóstica, Facultad De
Química, Universidad Autónoma De Querétaro, Querétaro,
México
Brenda Ugalde-Villanueva
Maestría En Química Clínica Diagnóstica, Facultad De
Química, Universidad Autónoma De Querétaro, Querétaro,
México
Alma Delia Bertadillo-Jilote
Maestría En Química Clínica Diagnóstica, Facultad De
Química, Universidad Autónoma De Querétaro, Querétaro,
México
David Gustavo García-Gutiérrez
Maestría En Química Clínica Diagnóstica, Facultad De
Química, Universidad Autónoma De Querétaro, Querétaro,
México
Marco Antonio Meraz-Ríos
Departamento De Biomedicina Molecular, Centro De
Investigación Y De Estudios Avanzados Del Instituto
Politécnico Nacional (CINVESTAV-IPN), Ciudad De México
Karla Isabel Lira-De León
Maestría En Química Clínica Diagnóstica, Facultad De
Química, Universidad Autónoma De Querétaro, Querétaro,
México

Page | 109
 Chapter - 6
Lung Cancer and Tobacco Smoking: Changes in Epigenetic
DNA Methylation
Diana Monserrat Aguilar-Beltrán, Brenda Ugalde-Villanueva, Alma Delia Bertadillo-
Jilote, David Gustavo García-Gutiérrez, Marco Antonio Meraz-Ríos and Karla Isabel
Lira-De León

Abstract
Nicotine addiction is the major cause that individuals persist in tobacco
smoking, and this persistent tobacco smoking contributes too many diseases.
Tobacco smoking is recognized as the leading cause of preventable death
worldwide, unfortunately its consumption has increased especially in young
people. In addition, tobacco smoking is associated with chronic pathologies
such as: chronic obstructive pulmonary disease, cardiovascular diseases,
cerebrovascular diseases and lung cancer, the latter being the one with the
highest incidence in relation to tobacco consumption, with 87% of cases and
82% of deaths from lung disease were due to its consumption. One of the
mechanism to develop lung cancer is due to changes in the regulation of gene
expression, particularly epigenetic changes such as DNA methylation, which
refers to the addition of a methyl group in the cytosine paired with a guanine,
which are generally enriched in the promoter regions of the genes. It is known
that this mechanism when presented in an aberrant manner may have direct
implications on the development of carcinogenesis. This review focuses on
analyzing the data that exists in the literature that associate Tobacco smoking
as one of the main risk factors in aberrant epigenetic regulation and
particularly in DNA methylation, whose changes appear in the earliest events
of the process of lung cancer development and evaluate DNA methylation as
one of the most promising biomarkers for the diagnosis of this disease.
Keywords: methylation, epigenetics, tobacco smoking, lung cancer
1. Introduction
Tobacco is a legal product whose exposure, increases the mortality of its
consumers up to 50%. The World Health Organization (WHO) has listed
smoking as the leading cause of preventable death in the world. Smoking is
associated with diabetes, respiratory and cardiovascular diseases, and cancer.

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Lung cancer (LC) is the most common disease worldwide, both in terms of
incidence and mortality, these data are closely related to smoking patterns [1],
Because in early stages of LC their course is asymptomatic, patients go to
medical review at late stages, when they present symptoms such as cough,
dyspnea, dysphonia, hemoptysis and chest pain, characteristic of advanced
stages (III or IV), which unfortunately, it implies a survival of approximately
only 5 years [2]. Tobacco smoke has hazardous effects on exposed populations,
including induction of a large amount of mutations in the genome and several
epigenetic abnormalities, initiation of chronic inflammation, and facilitating
immune escape of transformed cells3. The epigenetic abnormalities include a
deregulation of the methylation in oncogenes which subsequently causes their
overexpression, and conversely, a silencing of both tumor suppressor and
DNA repair genes, this is done through a high level of methylation in their
promoters. These changes can also persist for years after quitting smoking [3,
4]
. There are several reports, using different molecular analyzes and techniques,
that have managed to link smoking with different patterns of aberrant
methylation for LC [5].
2. Tobacco smoking
According to the WHO, smoking causes the death of five million direct
consumers and more than 600,000 indirect or exposed to smokers of tobacco
smoke [6, 7]. Tobacco, an agricultural product processed from the leaves of
Nicotiana tabacum, a plant native to America, was formerly used for a curative
and ceremonial purpose, which has changed completely and has now become
a regular consumption for some people [7]. The most common way in which
tobacco smoke is ingested is through cigarettes [8], the combustion of a
cigarette can generate more than 4000 different chemical products, of which
it is known that at least 250 of them are harmful to health and approximately
60 can cause cancer [9, 10]. Some of these chemicals are: tar, lead, arsenic,
ammonia, carbon monoxide, nicotine (substance responsible for tobacco
addiction), and many others. Tobacco alterations, costs to the world economy
about $ 200 billion each year, both in health services, as well as in
environmental and social damages [11].
Two types of smoke currents can be distinguished when consuming
tobacco; a main or particle phase, which is inhaled by the smoker and contains
25% of the total carcinogens such as nicotine and second a tar of lateral or
condensed stream, with the remaining 75% of products such as carbon
monoxide and tar derivatives, from the passive combustion of tobacco that
passes into the ambient air and is responsible for the occurrence of neoplasms
in passive smokers [12, 13].

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 The need to use tobacco, as shown in Figure 1, is due to the fact that
nicotine acts, in the brain, on the β subunit of the ionotropic acetylcholine
nicotinic receptor (nAch), inducing the release of neurotransmitters such as
dopamine and serotonin, which activate the hypothalamus, which in turn, has
under its influence the path of the system of emotional reactions and cerebral
reward: pleasure, excitement, relief of anxiety, vigilance, reduction of hunger,
increased cortisol, increased frequency and cardiac output, among other
effects [14, 15]. At the same time, a tobacco molecule called Mono-Amino
Oxidase Inhibitor (MAOI) blocks the enzyme MAO (Mono-amino-oxidase)
which is responsible for controlling the excess of dopamine, thereby
prolonging the action of it [16].
Figure 1. Effects of nicotine in the brain. The main active nicotine in
tobacco smoke acts on acetylcholine receptors, activating neurons that release
dopamine, while inhibiting the mono-amino oxidase responsible for
deactivating dopamine, which acts on the smoker's reward circuit, creating
addiction.

Fig 1: Effects of nicotine in the brain

In women, tobacco causes more severe damage than in men due to the
characteristics of the body and the female hormonal system, increasing the
risk of developing diseases of the respiratory, cardiovascular and different
types of cancer, such as breast and lung [17, 18].
3. Lung cancer
LC is a global problem and a public health issue worldwide, and are
closely related to smoking patterns [1]. The lung is a complex organ that
contains many different types of epithelial cells that are distributed in several
different regional microenvironments throughout the lung. LC begins in the
cells of the bronchi, bronchioles and alveoli, when the epithelial cells of the

Page | 113
pulmonary parenchyma have altered composition associated with an acute or
chronic lung injury that has the potential to develop LC [3]. The progression of
the LC goes from the interaction of the cancer cells with the cellular and
extracellular matrix environment. Genetic and epigenetic changes, such as
aberrant promoter methylation and alteration of the expression of microRNAs,
are the cause of the erroneous expression of collagen, proteoglycans, laminins,
proteases and integrins in the tumor microenvironment. In addition, signaling
processes modulate the conversion of fibroblasts to contractile fibroblasts that
are associated with cancer, tumor angiogenesis, immune system escapement
and the formation of a permissive niche for cancer progression [19]. There are
two main types of LC: small cell lung cancer, responsible for 15% of cases
and 85% non-small cell lung cancer, which is subdivided into three subtypes:
adenocarcinoma, squamous cell carcinoma and large cell carcinoma. This
classification is based on histological characteristics [20]. According to the
predominant location, adenocarcinoma is presumed to originate from distal
airways, while squamous cell carcinoma develop from central airways [21].
Unfortunately early stages of LC are asymptomatic and the diagnosis
occur in late stages of the disease, with a consequent bad prognosis, whit a
survival of less than 5 years [2]. The diagnosis includes the staging of the LC
to describe its size, position and whether it has spread. A number/letter system
is used to describe the stages from “I” through “IV”, according to the TNM
System neoplasm staging method, developed by the American Joint
Committee on Cancer in collaboration with the International Union against
Cancer. The “T” category describes the primary tumor site, the “N” category
describes the regional lymph node involvement and the “M” category
describes the presence or otherwise of distant metastatic spread. A patient with
a lower stadium of progression has a better prognosis [21].
4. DNA damage
Tobacco smoking is the main etiologic agent of lung cancer, and
approximately 80 to 90% of cases are related to this addiction. Since, only 10-
15% of active smokers will develop lung cancer, which suggests the
possibility of a specific susceptibility related to the individual's genetic
predisposition. The high variability of the tobacco as a cancer inducer, also
could be attributed to the variable content that cigarettes have of tar and to the
modifications made in the tobacco mixture of each geographical area, which
is done to make the inhalation of nicotine more pleasant; including the use of
filters that reduce the size of inhaled particles, which facilitates the deposition
of carcinogens at a deeper level of the cellular layers of the pulmonary
epithelium and causing that the DNA damage could be more permanent [22, 23].

Page | 114
 The mechanisms of how smoking affects DNA are becoming better
known. The tobacco components can induce the disruption of the helical
structure of DNA; in general, these ruptures result from the attack on the
deoxyribose sugar residues of the DNA, modifying the chemical base, forming
adducts, inducing cross-links between chains or even breaking the double
helix of the DNA. Single chain ruptures represent the initial damage of DNA
and are often used as a biomarker of exposure to carcinogens such as tobacco
smoke. Double strand ruptures can create apurinic or pyrimidine sites for
damage to phosphodiester bonds that bind nucleotides or for elimination of
corresponding bases [24] and are considered biologically more relevant, since
they can lead to chromosomal translocation, deletion or insertion and it is
known to be more conducive to the development of cancer, since if the DNA
is not repaired properly, these changes can severely alter the genetic structure
and expression [25].
On the other hand, nicotine can modulate gene expression directly,
through the binding and activation of acetylcholine receptors by increasing the
intracellular calcium leading to downstream activation of cAMP, a second
messenger involved in the transcription of genes that they mediate various
cellular responses such as inhibition of cell proliferation, innate and adaptive
immunological responses and cell cycle progression (G0 stage cells have a
higher cAMP content) [4, 26]. Similarly, cAMP can modulate gene expression
indirectly, regulating the activity of binding factors or DNA transcription
factors. It has been shown, for example, that the condensate of cigarettes
produced by smoking increases the expression of Sp1 and it’s binding to DNA
in lung epithelial cells. Sp1 is a transcription factor that is involved in a wide
variety of essential biological processes such as: preventing de novo
methylation in early embryogenesis, a phenomenon that has proven important
in cell growth, differentiation, apoptosis and carcinogenesis. Having high
levels of Sp1 protein is considered a negative prognostic factor in patients who
have lung cancer [27].
5. Epigenetic processes: methylation
Transcriptional regulation of genes involved in cell proliferation and
death is important in the onset and progression of cancer; It has been shown
that not only genetic mutations contribute to the development of cancer, but
there are also alterations that do not involve changes in the DNA sequence and
that actively participate in the carcinogenic process [28]. These variations are
known by the term "epigenetic variations" and lead to alterations that can be
transmitted to daughter cells (fortunately it is known that some of these
transformations can be reversed) [29]. These modifications include: DNA/RNA

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methylation, histone modifications, nucleosome positioning and altered
expression of non-coding RNAs, all these modifications can be identified as
markers of cancer onset and progression [30], since they decide which functions
are active and which are inactive in each specific tissue.
5.1 S-adenosyl methionine
The degree of expression of a gene is given in part by epigenetic
mechanisms, and these can be altered as a result of environmental influences,
such as nutritional status; This is especially relevant in complex diseases such
as cancer, diabetes or cardiovascular disease. Methyl groups are acquired
through diet and are donated to DNA through the folate and methionine
pathway. Dietary factors are essential to form S-adenosylmethionine (SAM),
which is the largest biological donor of methyl groups. The formation of SAM
involves three interrelated biochemical pathways: the folate cycle, the
methionine cycle and the transsulfuration pathway.
Folate cycle: Folate is a water-soluble vitamin B whose active form is
the tetrahydrofolate (THF), and it is used in the formation of purines and
pyrimidines, and in the remethylation of homocysteine to methionine. THF is
converted to 5-methyl-THF by the enzyme Methyl-tetrahydrofolate reductase
(MTHFR).
1. Methionine cycle
Methionine is an essential sulfur-containing gluconeogenic amino acid
that is converted to S-adenosylmethionine (SAM) by the enzyme methionine
adenosyltransferase (MAT) using ATP as co-substrate. SAM’s methyl group
is then transferred to a legion of different substrates, such as DNA, RNA,
glycine, and guanidinoacetate. When SAM donates its methyl, S-adenosyl-
homocysteine (SAH) is formed, and after the release of the adenosyl group,
we obtain homocysteine. The latter can be remethylated to methionine by
donating the methyl group of 5-methyl-THF, or it can be catabolized to
cysteine through the transsulfuration pathway.
2. Transsulfuration pathway
Homocysteine can be converted into cysteine and α-Ketobutyrate, which
after several reactions will be converted into pyruvate and succinyl CoA
necessary in gluconeogenesis. A decrease in the production of 5-methyl-THF
is associated with high levels of homocysteine and DNA hypomethylation.
The SAM/SAH ratio defines the methylation potential of a cell.
Hyperhomocysteinemia decreases this ratio and decreases the methyler
potential (Guardiola, Vallvé, Zaina, & Ribalta, 2015).

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 SAM-dependent methylation is essential for many biological processes
and its stationary level has been accepted as a critical marker of genomic
methylation; being high in quiescent cells and low in proliferating cells [31].
5.2 Histone methylation
At least eight types of post-translational modifications have been found
in histones that participate in the chromatin compaction state; the best known
are acetylation, methylation, phosphorylation and ubiquitination. Methylation
and demethylation, are catalyzed by the methyltransferase enzymes (HMTs)
and demethylases (HDMTs) respectively, and can be: mono, di or tri methyl
lysine (K-me1, K-me2, K-me3), or mono or dimethyl arginine (R-me1, R-
me2). The different histone modifications act in combination to form the so-
called "histone code", which can initiate biological responses such as
activation or repression of transcription. In general, transcriptionally inactive
genes have low levels of acetylation, methylation and phosphorylation, while
transcriptionally active chromatin has elevated levels of acetylation [32, 33].
5.3 Methylation of non-coding RNA (ncRNA) DNA regions
Only less than 2% of the human genome is used to encode proteins, the
majority of the mammalian genome is actively transcribed to produce large
amounts of non-coding RNA (ncRNA). Non-coding RNAs are functional
RNA molecules that, although not coding for proteins, are key in gene
regulation. The mRNAs do not produce changes on the DNA molecule unlike
histone methylation and DNA, but are responsible for controlling gene
expression in a post-transcriptional way by binding to target messenger RNAs
(mRNA), and thus promoting or their degradation, or inhibiting its translation,
preventing in both cases gene expression. The two main types of ncRNA are
micro RNA (miRNA) and long non-coding RNA (lncRNA). The miRNAs (22
nucleotides) are a class of recently discovered and well characterized ncRNAs,
they are responsible for fine-tuning and directing the translation of up to 60%
of the protein coding genes in humans, usually in the cytoplasm, and is often
deregulated in different types and subtypes of cancer. The lncRNA (200
nucleotides) also function as genetic regulators, but basically within the cell
nucleus. Although many studies have shown the deregulation of the
expression of ncRNA in several diseases, the epigenetic regulation of ncRNAs
remains poorly understood in different types of cancer and cancer subtypes.
Generalized epigenetic alterations such as hypermethylation of ncRNA
promoters that cause transcriptional inactivation in different types of cancer
have been observed, for example in breast cancer; Unlike genes that encode
proteins, most epigenetically poorly regulated ncRNAs were shared between

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breast cancer subtypes, but a subset of corresponding transcriptomic and
epigenetic changes occurred in a specific subtype mode [31, 34].
5.4 DNA methylation
DNA methylation is one of the most studied modifications, because of
their more direct effects on gene expression [35]. As shown in Figure 2,
methylation is a mechanism that is produced by adding a methyl group (CH3-)
to the DNA; the most widely differentiated process that represents the greatest
epigenetic modification of the human genome is the covalent addition of the
methyl group on carbon 5 of the cytosine pyrimidine ring, resulting in 5-
methylcytosine (5-mC). These methyl groups project into the major groove of
the helical structure of DNA and inhibit transcription [36]. These modifications
also participate in the protection of genome integrity and in the regulation of
gene expression, contributing with different epigenetic alternations [37], such
as chromosomal inactivation or instability usually associated with
hypomethylation, in the maintaining the silencing of certain genes during cell
differentiation and in the case of hypermethylation collaborates with the
inactivation of chromatin, specific tissue gene expression, genetic imprinting,
or silencing of transposable elements to avoid the generation of mutations.
This makes methylation a key player with an important role in the cell cycle
of healthy or cancer cells [31, 35].
Figure 2. Methylation of cytosine to 5-methylcytosine. Addition process
of a methyl group at position 5 of the cytosine that results in 5-methylcytosine
(5-mC), which can be projected into the major grooves of the DNA inhibiting
transcription.

Fig 2: Methylation of cytosine to 5-methylcytosine

In human DNA, 5mC, is found in approximately 1.5% of somatic cells,
5mc is produced almost exclusively in the context of symmetric methylation,
in which a cytosine nucleotide is located next to a guanine (CpG sites). CpG
dinucleotides are usually distributed heterogeneously throughout the genome,

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but are more concentrated in areas of the genome known as CpG Islands
(CGIs), which are usually associated with the promoter region of genes. The
methylation of CGIs has been associated with the repression of genes because
the presence of methylated cytosine produces a conformational change in the
double strand of DNA that makes it difficult to bind transcription factors and
even transcription itself. Gene promoters are normally non-methylated and
play a key role in controlling gene regulation [4, 36].
The DNA methylation profile and regulation of gene expression are
specific to each cell and tissue; the methylation pattern is established during
the embryonic development of mammals, while the zygote is formed, a non-
methylation pattern of the germ cell DNA takes place and the methylation
begins again during the necessary cell divisions of development ("epigenetic
reprogramming"). During this cellular development in vertebrates, DNA
methylation patterns must be established, maintained and eliminated. In
somatic cells it is enzymatically controlled by methyl transferases (DNMTs);
these are the only enzymes that have been shown to mediate the transfer of a
methyl group of SAM to cytosine. Without the DNMT's, the replication
machinery itself would produce non-methylated daughter chains, over time, it
would lead to passive demethylation [38]. At least three DNMT´s are involved
in DNA methylation and can be divided into two categories (see Figure 3): de
novo DNMT´s (DNMT3A and DNMT3B) which methylated non methylated
DNA; and maintenance DNMT´s (DNMT1) responsible for the
hemimethylated DNA, associated with DNA replication, to ensure that the
same methylation patterns are transmitted to daughter cells [32].
Figure 3. Role of methyltransferases. It shows the process of
methyltransferase enzymes (DNMT's), can mediate the cytosine methylation
patterns, at least three of them are involved in this process; DNMT3A and
DNMT3B methylated unmethylated DNA (de novo methylation) and DNMT1
methylated hemimethylated DNA (maintenance methylation).

Fig 3: Role of methyl transferases

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 Aberrant changes in methylation can suppress transcription, at least by
two known mechanisms: the first one is due to the presence of the methyl
group in a specific CpG that can directly block the recognition of DNA by
transcription factors and alternatively, some proteins (MBD, methyl binding
domain) can preferentially bind to methylated DNA, blocking access to the
transcription factor; Second, they indirectly do so by promoting the "closed or
compact" chromatin structure, preventing the union of transcriptional
machinery. As a consequence of these changes, two phenomena can occur:
1. Hypermethylation of promoter sequences, which leads to
transcriptional silencing of tumor suppressor genes and genes
involved in cell cycle control, DNA repair and apoptosis
2. Hypomethylation with the consequent overexpression of certain
proteins involved in the processes of invasion and metastasis [36, 39]
6. Lung cancer and DNA methylation
Although somatic genetic aberrations play an important role in the
carcinogenesis of lung cancer, epigenetic alterations are more frequent than
somatic mutations [40]. Therefore, the goal of new cancer research is to
understand the how? And the why? Of the onset and tumor progression.
In 2014 Nawaz et al. developed a specific multiple PCR of methylation
for the early detection of non-small cell lung cancer, to demonstrate the
silencing of tumor suppressor genes through DNA methylation, the sensitivity
of the assay was 87% and with a specificity of 94% and when related to
smoking showed a significant difference in more than one of the genes (out of
six they tested) for this activity [41].
The group of Gao et al. in 2016 examined whether smoking could alter
methylation of genes at lung cancer risk loci identified by genome-wide
association studies. DNA methylation levels of 2854 CpG candidates
corresponding to 75 genes were measured by the Illumina Infinium Human
Methylation 450 Beadchip array in whole blood samples. As a result, 13 CpG
sites were confirmed as significantly smoking-related loci, one site was
previously identified within the KLF6 gene, while 12 novel sites were located
in 7 genes: STK32A, TERT, MSH5, ACTA2, GATA3, VTI1A and CHRNA5.
Which may indicate that lung cancer susceptibility genes might be regulated
by methylation changes [42]. In the same line of research, Stueve et al. in 2017
identified seven hypomethylated CpGs associated with tobacco use using the
Infinium Human Methylation Chip, suggesting that biomarkers in blood may
reflect changes in target tissue [43].

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 As we mentioned at the beginning of this chapter, early diagnosis is
crucial for a better prognosis of the evolution of LC. Therefore, liquid biopsy
is currently offered as a tool to help diagnose and prognosis of these patients.
Liquid biopsy makes use of cells, nucleotides and proteins released from
tumor cells into body fluids. Methylation of circulating tumor DNA (ctDNA)
has gained increasing attention as biomarkers LC, ctDNA mostly results from
apoptosis and necrosis of primary and metastatic tumor [44]. However,
circulating cell-free DNA (cfDNA) is also present in body fluids, cfDNA is a
mixture of single or double-stranded DNA in circulation released from
different tissues including tumor, worth mentioning that is difficult to separate
ctDNA from cfDNA originated from non-cancer tissues. For this reason, it's
critical the selection of control group and target genes in a clinical trial. The
cfDNA can be isolated from both plasma and serum, but in the latter there is
more abundant [30, 45]. In non-small cells of LC, CGIs hypermethylation has
been associated with cigarette smoking. On the other hand, DNA
hypomethylation in repetitive sequences occurred in early stage of squamous
cell of LC, and it has been observed that individuals with hypomethylation in
repetitive element has a high risk of developing cancer [46]. Methylation occurs
at early stage of carcinogenesis (Figure 4), and has become an attractive
biomarker for cancer screening and early detection, especially for ctDNA
methylation with its convenience and non-invasion [47].
Figure 4. Aberrant DNA methylation in Lung Cancer. Methylation occurs
at early stage of carcinogenesis, were the hypermethylation in CGIs sites of
promoter sequences, which leads to transcriptional silencing of tumor
suppressor genes and genes involved: in cell cycle control, DNA repair and
apoptosis. This phenomenon does not occur in CGIs sites of promoter
sequences of normal cell tissue.

Fig 4: Aberrant DNA methylation in Lung Cancer

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 In 2020, Morrow et al. identified variable DNA methylation associated
with all-cause mortality in smokers with and without chronic obstructive
pulmonary disease (COPD). They evaluated predictive epigenetic marks of
smokers in peripheral blood that may allow a targeted risk stratification, in
particular they observed differential methylation of two replicated PIK3CD
sites were associated with lung function at enrollment. Also, these sites were
located in genomic regions of DNase hypersensitivity, highlighting the
potential regulatory relevance of these DNA methylation sites as biomarkers
of mortality [25].
Currently most of the aberrant methylation patterns can be reversed using
demethylations agents (chemotherapeutic agents), for this it is necessary to
identify the changes occurred in the genes, which is achieved through analysis
of gene expression, by sequencing techniques, PCR, for treatment with
bisulfite to DNA or pyrosequencing [48].
In conclusion, methylation studies and its changes in DNA could be used
as new markers of cancer, especially in the early stages for smokers, since
non-invasive analysis can provide an early diagnosis and its regulation could
be an efficient and less traumatic treatment for the patient. Although the
growing evidence suggests that epigenetic aberrations contribute to the
development and progression of cancers such as lung cancer, the possible
reversal of this process constitutes a new target, very attractive for the
development of therapeutic strategies that involve reactivation or re-silencing
of the specific genes that are deregulated.
Conflict of interests
The authors declare that they have no real or potential conflict of interest
with this article.
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