Determining the new type of mutation in EGFR gene prevalent in

Bengali population and assessing its frequency

Introduction:
Lung cancer is the most common cancer diagnosed worldwide. It is also the foremost
contributor to cancer-related mortality, resulting in 1.38 million cancer deaths per year
worldwide. Lung cancer accounts for more deaths than any other type of cancer. Several
epidemiological observations performed across varied demographic cohorts in India confirm
the significant burden of lung cancer in India, contributing significantly toward the cancer
morbidity and mortality. It kills 1.76 million people every year. In India, with a mortality rate
5/100,000, it is responsible for 63,475 deaths annually. It is responsible for 7.5% of cancer
Disability-Adjusted Life-Years (DALY) in the country. Out of 27 population-based cancer
registry, 10 in the country have recorded lung ca as the most common cancer in males. In males,
it is the most common cancer, with 10.4% and in females, it stands at seventh place, with 4.4%
contribution to cancer DALY.
Being a cancer with high fatality rate, there is need to have early diagnosis as the main
stay of treatment. In fact, the primary need might be to understand its epidemiology in Indian
population to contain it. Unfortunately, cancer registries are mostly urban in nature, thus not
being able to deliver a true picture of the rural parts. Few studies earlier linked smoking and
air pollution with this cancer. In addition, economic condition, alcohol and diet were also
claimed to have some important role in its distribution.
As a single etiologic agent, tobacco is by far the most important risk factor in the
development of lung cancer. It is estimated that around the world, 80% of lung cancer cases in
men and 50% in women each year are caused by smoking. The abundance of evidence of dose-
response relationship and biological plausibility overwhelmingly supports the existence of a
causal relationship between smoking and lung cancer. The same relationship exists between
passive smoking (so-called second-hand smoke) and lung cancer.
Outdoor or indoor air pollution is a significant environmental risk factor for lung
cancer; long-term exposure to polluted air caused by factories and automobiles, cooking fumes,
or formaldehyde from indoor decoration definitely increases the risk of lung cancer. Early
ecologic studies found that more than 50% of lung cancer occurred in urban areas, which is
most probably more from polluted air from industrial sources and vehicle exhaust than in rural
areas. A series of case control and cohort studies found a notable association between lung
cancer and air pollution with adequate adjustment for tobacco use and other potential risk
factors.
The International Agency for Research on Cancer has identified 12 occupational
exposure factors as being carcinogenic to the human lung (aluminum production, arsenic,
asbestos, bis-chloromethyl ether, beryllium, cadmium, hexavalent chromium, coke and coal
gasification fumes, crystalline silica, nickel, radon, and soot). Asbestos is a well-established
occupational carcinogen and refers to several forms of fibrous, naturally occurring silicate
minerals; exposure to asbestos at high levels can cause lung cancer and mesothelioma. From
cohort studies on cancer mortality among workers exposed to asbestos in China, a significantly
elevated meta–standard mortality ratio for lung cancer is 4.54, with 95% CI of 2.49 to 8.24.
Concurrent smoking and asbestos exposure are synergistic and result in increased cancer
incidence.

Hypothesis: Epidermal Growth Factor Receptor (EGFR) exon 20 insertion (ex20ins)

mutations occur in ~2–3% of all non-small cell lung cancer (NSCLC) cases, representing ~10–
12% of all cancers with documented EGFR mutation. These mutations are the third most
common EGFR mutation subtype after the common sensitizing EGFR mutations, i.e. the exon
19 deletions and exon 21 L858R mutation. In Bengali population, a large number of patients
have shown an altered RFLP pattern, when tested against the EGFR gene by using Hind III
restriction endonuclease. It is likely to be another deletion mutation in the exon 18, that is
causing an immature termination of the polypeptide, which is very much frequent in Bengali
population causing lung cancer.

Objectives:
Primary objective of the study -
Determination of the new type of mutation resulting in premature chain termination and
causing lung cancer in Bengali population in India.
Secondary Objective -
Determining the percentage of lung cancer patients in Bengali population showing this
type of mutation.
Experimental designs:
1) There will be a study consisting of thousand patients with diagnosed lung cancer from five
government hospitals, and assessment of the presence of the mutation in the agarose gel
pattern of RFLP by the restriction endonuclease enzyme Hind III in the EGFR gene, from
the tumour tissue.
2) Then there will be a sanger sequencing of the 18th exon region of the EGFR gene from
patients showing altered RFLP pattern.
3) This sequence obtained from sequencing will be tested by using sequence alignment in
BLAST database.
4) Presence of the particular mutation will be further tested through DNA micro array.
5) The deformed protein will also be tested by SDS page and subsequent western blotting.
6) Further population study for counting the percentage of that specific mutation causing lung
cancer in Bengali population will be assessed by ARMS PCR by using common primer and
ARMS primers.

Conclusion:
1) Different mutation, causing a new variety will be identified among patients with lung
cancer.
2) Drug design and targeted therapy to correct mutation should be possible.
3) The frequency of this type of mutation in Bengali population would be revealed.