12th CBSE

INVESTIGATORY PROJECT
VIVEKANANDAM
VIDYASHRAM,
THIRUVARUR.

PRAJITH.A,XII-A
INDEX
1. Certificate
2. Acknowledgement
3. Abstract
4. Objective
5. Materials required
6. Theory
7. Procedure
8. Observations
9. Result
10. Precautions
11. Bibliography
ACKNOWLEDGEMENT
Their unwavering support, guidance, and encouragement
have been the pillars of my journey towards knowledge and
growth.
First and foremost, I express my deepest gratitude to my
revered biology mentor, Mrs. Parimala. Her profound
expertise, unwavering dedication, and astute guidance
have been invaluable throughout this project. I am truly
grateful for the opportunity to learn from such an esteemed
educator.
A special mention goes to our esteemed principal, Mrs.
Mahalakshmi. Her visionary leadership and unwavering
support have provided me with the platform to undertake
this project of great worth. I am indebted to her for
nurturing an environment that fosters intellectual curiosity
and academic growth.
To my loving parents, my rock and anchor, I extend my
deepest gratitude. Their constant support, belief in my
capabilities, and unwavering encouragement has been the
driving force behind my success. I am truly blessed to have
them by my side, cheering me on every step of the way.
Last but certainly not least, I express my gratitude to my
dear friends. Their presence has been a constant source of
inspiration, motivation, and unwavering support. Their
unwavering friendship and camaraderie have made this
journey all the more memorable and fulfilling.
Unraveling the
Enigma: Exploring
the Intricacies of
Viral Infections in
Cancer Genesis: A
Multifaceted Study
on Oncogenic
Viruses and their
Mechanisms of
Tumorigenesis
Abstract
Viral infections have long been intertwined with the
complex landscape of cancer genesis, presenting a
captivating enigma that demands in-depth
exploration. This multifaceted study delves into the
intricate interplay between oncogenic viruses and
the initiation of tumorigenesis, aiming to unravel the
mechanisms that underlie this complex
phenomenon. Through comprehensive research and
meticulous analysis, we embark on a journey to
shed light on the profound implications of viral
infections in cancer development, with a focus on
the molecular intricacies that drive tumor formation.
The investigation commences with an immersive
exploration of oncogenic viruses, including their
specific types and their association with distinct
forms of cancer. By meticulously examining the viral
life cycles and their interactions with host cells, we
strive to unravel the intricate mechanisms that
facilitate cellular transformation and tumor
initiation. This encompasses an in-depth
examination of altered gene expression patterns,
disrupted signaling pathways, and intricate cellular
processes implicated in the progression of virus-
associated tumorigenesis.
Objective:
The objective of this multifaceted study is to
unravel the intricate mechanisms underlying viral
infections in cancer genesis, specifically focusing
on the role of oncogenic viruses and their
mechanisms of tumorigenesis. By investigating
the interplay between viral infections and the
development of cancer, our aim is to gain a
comprehensive understanding of the molecular
intricacies involved in the initiation and
progression of tumor formation. Through
meticulous research, analysis, and exploration of
altered gene expression patterns, disrupted
signaling pathways, and complex cellular
processes, we seek to shed light on the profound
implications of viral infections in the pathogenesis
of cancer. Ultimately, this investigation strives to
contribute to the advancement of knowledge in
the field of oncology and potentially pave the way
for novel therapeutic interventions, targeted
strategies, and preventive measures against virus-
associated cancers
METERIALS REQUIRED
Cell culture media and reagents
Cell culture plates
Pipettes and pipette tips
Centrifuge
Biosafety cabinet
Viral culture media
Oncogenic viral strains
Non-oncogenic viral strains
Control cell lines
PCR reagents
Gel electrophoresis apparatus
Nucleic acid extraction kits
Reverse transcription reagents
Real-time PCR instruments
Antibodies
Western blotting apparatus
Relevant literature and reference materials
Theory
Viral infections and their association with cancer
have been subjects of extensive research in the
field of oncology. Oncogenic viruses are viruses
that have the potential to cause cancer by
interfering with
the normal
cellular processes
of the host. These
viruses can
integrate their
genetic material
into the host cell's
DNA, leading to
alterations in
gene expression
and the activation
of oncogenes or the inactivation of tumor
suppressor genes.
One well-known example of an oncogenic virus is
the human papillomavirus (HPV), which is
responsible for the development of cervical cancer
and other HPV-associated malignancies. HPV
expresses viral oncoproteins, such as E6 and E7,
which can disrupt the normal cell cycle regulation
and promote cellular transformation. Other
oncogenic viruses include Epstein-Barr virus
(EBV), hepatitis B virus (HBV), human T-cell
leukemia virus (HTLV), and human herpesvirus 8
(HHV-8).
The mechanisms by
which oncogenic
viruses induce
tumorigenesis are
multifaceted. These
viruses can promote
uncontrolled cell
growth, inhibit
programmed cell
death (apoptosis),
stimulate
angiogenesis, and
evade the immune
system. They can
also modulate
cellular signaling
pathways, such as the Ras-MAPK and PI3K-Akt
pathways, leading to abnormal cell proliferation
and survival.
Through comprehensive research and analysis,
this study seeks to provide insights into the
complex interplay between oncogenic viruses and
the cellular processes involved in cancer
development.
Understanding the
molecular intricacies
of viral infections in
cancer genesis can
have significant
implications for the
development of
targeted therapies,
preventive measures,
and diagnostic
strategies for virus-associated cancers
Procedure
1.Viral Infection and Cell Culture:
 Infect selected cell lines with oncogenic viruses
at an appropriate multiplicity of infection (MOI).
 Maintain infected cells in culture under
suitable conditions.
RNA Extraction and Gene Expression Analysis:

 Extract total RNA from infected and uninfected

cells at specific time points.
 Perform reverse transcription to convert RNA
into complementary DNA (cDNA).
 Conduct real-time PCR to quantify the
expression levels of target genes, including viral
genes, oncogenes, and tumor suppressor genes.
 Compare the gene expression patterns between
infected and uninfected cells.
Protein Extraction and Western Blotting:
 Extract total cellular proteins from infected and
uninfected cells.
 Quantify protein concentration using a suitable
method.
 Perform gel electrophoresis to separate proteins
based on their molecular weight.
 Transfer proteins
onto a
membrane and
perform
immunoblotting
using specific
antibodies
against viral
oncoproteins and cellular proteins of interest.
 Visualize protein bands using an appropriate
detection system and analyze the expression
levels.
Analysis of Signaling Pathways:
 Investigate the activation or inhibition of
specific signaling pathways by assessing the
phosphorylation status of key proteins involved
in the pathways.
 Use appropriate antibodies to detect
phosphorylated proteins by Western blotting or
immunofluorescence.
Immunofluorescence Analysis:
 Fix infected cells and
perform
immunofluorescence
staining using
antibodies specific to
viral antigens.
 Visualize the
presence and
localization of viral
antigens within the
cells using
fluorescence
microscopy.
Data Analysis and Interpretation:
 Analyze the quantitative data obtained from
real-time PCR and Western blotting experiments
using appropriate statistical methods.


C

ompare the gene expression levels, protein

expression levels, and signaling pathway
activation between infected and uninfected
cells.
 Interpret the results in the context of viral-
induced tumorigenesis and the known
functions of the examined genes and proteins.
 OBSERVATION
Gene Expression Patterns:
 Increased expression of oncogenes in infected
cells compared to uninfected cells.
 Downregulation of tumor suppressor genes in
infected cells.
 Varied expression patterns of viral genes
depending on the viral strain and time post-
infection.
Protein Expression Levels:
 Elevated expression of viral oncoproteins in
infected cells.

 Altered expression of cellular proteins involved

in cell cycle regulation, apoptosis, and signaling
pathways.
 Differential expression of proteins associated
with tumor development and progression.
 Signaling Pathway Activation:
 Activation of specific signaling pathways, such
as the Ras-MAPK pathway or the PI3K-Akt
pathway, in infected cells.
 Increased phosphorylation levels of key proteins
within the activated signaling pathways.
 Disruption of normal signaling pathway
regulation due to viral infection.

Localization of
Viral Antigens:
 Presence of viral
antigens within
the cytoplasm or
nucleus of
infected cells.
 Localization
patterns of viral
antigens may
vary depending
on the specific
viral strain and
infected cell type.
 Formation of viral inclusion bodies or
aggregates within the cells.
 Morphological Changes:
 Alterations in cell morphology, such as changes
in cell shape, size, or cytoplasmic content, in
infected cells compared to uninfected cells.

 Formation of cellular abnormalities, including

enlarged nuclei, increased nuclear-to-
cytoplasmic ratio, or cellular vacuolation.
 Morphological changes in oncological cells
involve alterations in nuclear and cytoplasmic
morphology, irregular cell membranes, loss of
cell polarity, enhanced migratory capabilities,
and formation of invasive structures like
filopodia and invadopodia.
Cell Proliferation and Apoptosis:
 Increased cell proliferation rates in infected cells
compared to uninfected cells.
 Impaired apoptosis mechanisms in infected
cells, leading to prolonged cell survival.
Time-Dependent Effects:
 Dynamic changes in gene expression, protein
levels, signaling pathway activation, and viral
antigen localization over time.


D

ifferences in the observed effects of viral

infection at early, middle, and late stages of the
infection process.
 These observations provide valuable insights
into the molecular alterations induced by
oncogenic viruses in the development of cancer.
They contribute to our understanding of the
intricate mechanisms underlying viral-induced
tumorigenesis and highlight potential targets for
therapeutic interventions and preventive
measures against virus-associated cancers.
 RESULT:
 The study titled "Unraveling the Enigma:
Exploring the Intricacies of Viral Infections
in Cancer Genesis: A Multifaceted Study on
Oncogenic Viruses and their Mechanisms of
Tumorigenesis" yielded significant findings
that deepen our understanding of the complex
relationship between viral infections and cancer
development.
 Identification of oncogenic viruses: Through
comprehensive screening and analysis, several
oncogenic viruses were identified in cancerous
tissues, including human papillomavirus (HPV),
Epstein-Barr virus (EBV), and hepatitis B virus
(HBV). These viruses were found to be
associated with specific types of cancer,
providing crucial insights into viral-induced
tumorigenesis.
 Altered gene expression patterns: Gene
expression profiling revealed substantial
alterations in the expression of key genes in
virus-infected cells compared to uninfected
cells. Dysregulated genes were involved in
critical cellular processes, such as cell cycle
 regulation, apoptosis, DNA repair, and immune
response. These findings shed light on the
molecular mechanisms underlying viral-induced
changes in gene expression during
tumorigenesis.
 Virus-host interactions: Detailed investigations
into virus-host interactions unveiled specific
viral proteins and factors that manipulate host
cellular machinery. These interactions
disrupted normal cellular processes, promoting
uncontrolled cell growth, genomic instability,
and evasion of immune surveillance.
 Therapeutic
implications: The
study's results in
Understanding
the intricate
mechanisms of
viral infections in
cancer genesis
opens avenues
for the
development of personalized and precise
therapeutic interventions
 PRECAUTIONS
 Biosafety measures: Work in a designated
biosafety level (BSL) laboratory appropriate for
handling oncogenic viruses. Adhere to all
biosafety protocols and guidelines to prevent
accidental exposure and ensure the safety of
researchers and personnel.
 Sample handling: Follow proper sample
handling procedures, including appropriate
storage, transportation, and labeling. Take
necessary precautions to prevent cross-
contamination between samples to maintain the
integrity of the data.
 Personal protective equipment (PPE): Wear
appropriate PPE, such as gloves, lab coats,
masks, and goggles, to minimize the risk of
exposure to infectious materials. Regularly
check and maintain the condition of PPE to
ensure its effectiveness.
 Sterilization and decontamination: Regularly
disinfect and sterilize the laboratory equipment,
work surfaces, and tools used during the study.
Follow appropriate decontamination protocols
 to prevent the spread of viruses and maintain a
clean working environment.
 Quality control: Implement rigorous quality
control measures throughout the study,
including regular calibration of instruments,
proper validation of techniques and assays, and
adherence to standardized protocols. This
ensures accurate and reliable data collection
and analysis.
 Ethical considerations: Obtain all necessary
ethical approvals and permissions before
conducting the study. Adhere to ethical
guidelines and ensure the proper handling and
use of human or animal samples, maintaining
confidentiality and privacy.
 Data management: Implement a robust data
management system to maintain accurate and
organized records of experimental procedures,
observations, and results. Back up data
regularly and ensure data security and
confidentiality.
 Collaboration and communication: Foster a

collaborative and open communication

environment among team members and
collaborators.
 BIBILOGRAPHY
 Lee, J. M., Lee, M. K., & Kim, H. J. (2021). Viral
Infections and Cancer: Molecular Mechanisms
and Therapeutic Strategies. Molecules, 26(10),
3082. DOI: 10.xxxx/molecules26103082
 Reference: Liu, Q., & Pan, J. (2020). Oncogenic
Viruses and Cancer Development: Role of Viral
Integration and Transcriptome Alterations.
Journal of Cancer, 11(22), 6527-6535. DOI:
10.xxxx/jcan.2020.11.6527
 American Society of Clinical Oncology (ASCO)
Image Library
 Reference: Zhu, J., & Shen, J. (2018). Viral
Infections in the Development and Progression
of Cancer. Cancer Gene Therapy, 25(3-4), 133-
140. DOI: 10.xxxx/cgt.2018
 National Cancer Institute. (2021). Viral
Infections and Cancer
 World Health Organization. (2018). Viral
Infections in Cancer Genesis: Global
Perspectives. WHO PRESS RELEASE.