REVIEW OF RELATED LITERATURE

Cancers figure among the leading causes of morbidity and mortality worldwide, with
approximately 14 million new cases and 8.2 million cancer related deaths in 2012. The number
of new cases is expected to rise by about 70% over the next 2 decades. Among men, the 5
most common sites of cancer diagnosed in 2012 were lung, prostate, colorectum, stomach, and
liver cancer. Among women the 5 most common sites diagnosed were breast, colorectum, lung,
cervix, and stomach cancer. More than 60% of worlds total new annual cases occur in Africa,
Asia and Central and South America. These regions account for 70% of the worlds cancer
deaths. It is expected that annual cancer cases will rise from 14 million in 2012 to 22 within the
next 2 decades. (World Health Organization, 2015)
According to the Union for International Control (2013), cancer kills more people than
AIDS, malaria and TB combined globally. Not only does cancer has a huge impact on loss of life
but in the economy as well. In 2010, the disease costs an estimated $290 billion- $154 billion of
which were medical costs.
Cancer is a multi-step process in which cells undergo metabolic and behavioural
changes, leading them to proliferate in an excessive and untimely way. These changes arise
through modifications in mechanisms that control cell proliferation and lifespan, relationships
with neighbouring cells, and capacity to escape the immune system. Modifications that lead to
cancer include genetic changes that modify the DNA sequence. Another way to change the
programme of cells is to modify the conformation of chromatin, the structure that wraps up DNA
and regulates its access by DNA reading, copying and repair machineries. Such changes are
called epigenetic. Advances in cancer molecular biology also identify new ways to inhibit the
growth of cancer, leading to new, more selective and less toxic forms of cancer chemotherapy.
(Kanin, 2008)

Cancer is a disease that is often characterized by too little apoptosis. Apoptosis, or

programmed cell death, is a normal component of the development and health of multicellular
organisms. Cells die in response to a variety of stimuli and during apoptosis they do so in a
controlled, regulated fashion. Under normal circumstances damaged cells will undergo
apoptosis, but in the case of cancer cells mutations may have occurred that prevent cells from
undergoing apoptosis. In these cases there is no check on the cellular proliferation and
consequently the disease can progress to the formation of tumors. (Dash, ????)
In the last decade, basic cancer research has produced remarkable advances in the
understanding of cancer biology and cancer research. Among the most important of these
advances is the realization that apoptosis and the genes that control it have a profound effect on
the malignant phenotype. (Oxford, 1999)
Today, nanotechnology is operating in various fields of science via its operation for
materials and devices using different techniques at nanometer scale. Nanoparticles are a part of
nanomaterials that are defined as a single particles 1100 nm in diameter. From last few years,
nanoparticles have been a common material for the development of new cutting-edge
applications in communications, energy storage, sensing, data storage, optics, transmission,
environmental protection, cosmetics, biology, and medicine due to their important optical,
electrical, and magnetic properties. There are various types of nanoparticles reported in the
literature, e.g., metal nanoparticles, metal oxide nanoparticles, and polymer nanoparticles.
Among all these, metal oxide nanoparticles stand out as one of the most versatile materials, due
to their diverse properties and functionalities. Most preferentially, among different metal oxide
nanoparticles, zinc oxide (ZnO) nanoparticles have their own importance due to their vast area
of applications, e.g., gas sensor, chemical sensor, bio-sensor, cosmetics, storage, optical and
electrical devices, window materials for displays, solar cells, and drug-delivery. ZnO is an
attractive material for short-wavelength optoelectronic applications owing to its wide band gap

3.37 eV, large bond strength, and large exciton binding energy (60 meV) at room temperature.
As a wide band gap material, ZnO is used in solid state blue to ultraviolet (UV) optoelectronics,
including laser developments. In addition, due to its non-centrosymmetric crystallographic
phase, ZnO shows the piezoelectric property, which is highly useful for the fabrication of
devices, such as electromagnetic coupled sensors and actuators. (Hahn et. al., )
Zinc oxide (ZnO) nanopowders are available as powders and dispersions. These
nanoparticles exhibit antibacterial, anti-corrosive, antifungal and UV filtering properties. Zinc is a
Block D, Period 4 element while Oxygen is a Block P, Period 2 element. Some of the synonyms
of zinc oxide nanoparticles are oxydatum, zinci oxicum, permanent white, ketozinc and oxozinc.
Zinc oxide is used in the manufacture of rubber and cigarettes (used as a filter). Popularly
known calamine lotion is made out of zinc oxide powder. It is also used in a host of other
creams and ointments that are used to treat skin diseases and as an additive in the
manufacture of concrete. Ceramic industry has a number of uses for zinc oxide powder. It is
also used as an additive in food products such as breakfast cereals. Various paints use zinc
oxide as a coating agent. (Azonano, 2013)
ZnO nanoparticles, with their unique properties such as biocompatibility, high selectivity,
enhanced cytotoxicity and easy synthesis, may be a promising anticancer agent. Zinc, as one of
the major trace elements of the human body and co-factor of more than 300 mammalian
enzymes, plays an important role in maintaining crucial cellular processes including oxidative
stress, DNA replication, DNA repair, cell cycle progression and apoptosis. Thus, it is evident that
an alteration in zinc levels in cancer cells can cause a deleterious effect. Research has shown
that low zinc concentration in cells leads to the initiation and progression of cancer and high zinc
concentration shows toxic effects. Zinc-mediated protein activity disequilibrium and oxidative
stress through reactive oxygen species (ROS) may be the probable mechanism of this cytotoxic
effect. The selective localization of ZnO nanoparticles towards cancer cells due to enhanced
permeability and retention (EPR) effect and electrostatic interaction and selective cytotoxicity

due to increased ROS present in cancer cells show that ZnO nanoparticles can selectively
target and kill cancer cells, making them a promising anticancer agent. (Bischt et. al, 2016)
In a study by Albrecht et. al. in 2013, ZnO treatment caused a rapid induction of nuclear
condensation, DNA fragmentation, and the formation of hypodiploid DNA nuclei and apoptotic
bodies. The involvement of the essential effector caspase-3 in ZnO-mediated apoptosis could
be demonstrated by immunocytochemical detection of activated caspase-3 in RAW 264.7 cells.
ZnO specifically triggered the intrinsic apoptotic pathway, because Jurkat T lymphocytes
deficient in the key mediator caspase-9 were protected against ZnO-mediated toxicity whereas
reconstituted cells were not. ZnO also caused DNA strand breakage and oxidative DNA damage
in the RAW 264.7 cells as well as p47phox NADPH oxidase-dependent superoxide generation
in bone marrow-derived macrophages.
Studies have shown that ZnO NPs induce cytotoxicity in a cell-specific and proliferationdependent manner, with rapidly dividing cancer cells being the most susceptible, and quiescent
cells being the least sensitive. Clearly, the type of cell in question is important when considering
the cytotoxicity of ZnO NPs toward mammalian cells. Apoptosis is a key process in cancer
development and progression. The ability of cancer cells to avoid apoptosis and continue to
propagate is one of the fundamental hallmarks of cancer and is a major target of cancer
treatment. Apoptosis is controlled by a large number of genes acting as death switches. The
tumor suppressor gene p53 is regarded as the master guardian of the cell and is able to activate
cell-cycle checkpoints, DNA repair, and apoptosis to maintain genomic stability. In the presence
of DNA damage, p53 protein triggers cell-cycle arrest to provide time for the damage to be
repaired or for self-mediated apoptosis. The bax/bcl-2 protein ratio determines the life or death
of cells in response to an apoptotic stimulus. A higher level of bax/bcl-2 ratio decreases the
resistance to apoptotic stimuli, leading to apoptosis. The bcl-2 protein has an anti-apoptotic
effect, whereas the bax is known for pro-apoptotic activity. It has also been well documented

that signalling pathway leading to apoptosis involves the sequential activation of cysteine
proteases known as caspases. (Akhtar et. al, 2012)
Akhtar et.al showed utilized human liver cancer (HepG2) to explore the possible
molecular mechanisms of toxicity induced by ZnO NPs. They observed that the expressions of
both mRNA and protein levels of tumor suppressor gene p53 and proapoptotic genes (bax and
caspase-3) were upregulated while the expression of anti-apoptotic gene bcl-2 was
downregulated in cancer cells treated with ZnO NPs. In the presence of DNA damage or cellular
stress, p53 triggers cell-cycle arrest to provide time for the damage to be repaired or for selfmediated apoptosis. ZnO NPs significantly altered the oxidant/antioxidant status of human liver
cancer cells. ROS generation and membrane LPO were significantly higher, whereas
antioxidant molecule GSH and the activity of antioxidant enzymes SOD, CAT, GPx, and GR
were significantly lower in ZnO NP-treated cells. GSH, a ubiquitous and abundant cellular
antioxidant tripeptide, was found to be strongly depleted in response to NP treatment.
Mitjans and Vinardell in 2015 showed that zinc oxide NPs were used at a very low
concentration and were found to exhibit activity against HepG2 (liver cancer) and MCF-7 (breast
cancer) cancer cells in a dose-dependent manner. The results of these anti-proliferative studies
clearly demonstrate that treatments with NPs sensitize cancer cells. The degree of apoptosis
was found to be enhanced with an increase in the concentration of NPs, and a significant
concentration of NPs resulted in cell death in both cancer cell lines. In the study, quantitative
real-time PCR was utilized to analyze the mRNA levels of apoptotic markers (p53, Bax, bcl-2
and caspase-3) in HepG2 cells exposed to ZnO NPs at a concentration of 50 g/mL for 24 h.
The results showed that the mRNA levels of these apoptotic markers were significantly altered
in HepG2 cells due to ZnO NP exposure.
Gao et. al. in 2014 conducted a study on Zno nanoparticles treatment which induced
apoptosis by increasing intracellular ROS levels in Ltep-a-2 cells. Their results showed that
ZnO NPs imposed distinct toxic effect on LTEP-a-2 cells, and the declines in cell viability and

survival rate coincided with specific morphological changes and the occurrence of apoptosis.
Further exploration of the toxicological mechanism revealed that the increase in ROS
coincided with depletion of GSH in apoptotic cells, suggesting that oxidative stress may be
the primary toxicological mechanism of ZnO NPs in LTEP-a-2 cells. As a common
mechanism for NPs-induced cell oxidative damage, increased ROS generation has been
confirmed by in vivo and in vitro tests of a wide range of NPs species.