Oxidative stress and its implications in cervical cancer.pptx

M
Mahima GuptaStudent at Home
“OXIDATIVE STRESS AND ITS IMPLICATION IN
CERVICAL CANCER”
Submitted By
Mahima Gupta
Class: M.Sc. Biotechnology, IV Semester
Roll No.: 2110014155034
Under the supervision of
Prof. Monisha Banerjee
Institute of Advanced Molecular Genetics & Infectious Diseases (IAMGID)
ONGC Centre for Advanced Studies (OCAS)
University of Lucknow
Submitted to
Department of Biochemistry,
University of Lucknow
CONTENTS
 Introduction to cervical cancer
 Oxidative Stress and Cervical Cancer
 Antioxidant enzyme and their role in cervical cancer
 Methodology
Protein estimation
GSH estimation
Catalase estimation
SOD Activity
 Results
 Discussion
 Conclusion
 References
INTRODUCTION
Cervical cancer is the third most
common cancer among women in world.
In India , cervical cancer is the second
most common.
Cervical cancer is a type of cancer that
occurs in the cells of the cervix — the
lower part of the uterus that connects to
the vagina.
Major cause is Human papillomavirus
(HPV).
CELL TYPES OF CERVICAL CANCER
 Squamous cell :This type of cervical cancer begins in
the thin, flat cells (squamous cells) lining the outer part
of the cervix, which projects into the vagina. Most
cervical cancers are squamous cell carcinoma.
 Adenocarcinoma:This type of cervical cancer begins
in the column-shaped glandular cells that line the
cervical canal.
According to Globocan 2020 total cases of cervical cancer were 1393717 in
worldwide ( age group 5-74 years) among those 297119 were died.
 In India estimated number of prevalent cases (5 years) in 2020, females ages
5-74 is 1654540 and total estimated deaths in 2020 females ages 5-74 is 363844.
Prevalence of cervical cancer
RISK FACTOR
OXIDATIVE STRESS AND CERVICAL CANCER
 Oxidative stress is an imbalance
between the production of reactive
oxygen species (ROS) and the body’s
ability to detoxify them.
 ROS can cause damage to cellular
components such as DNA, proteins,
and lipids, leading to cellular
dysfunction and death.
 This process has been implicated in
the development and progression of
various cancer, including cervical
cancer.
ANTIOXIDANT ENZYME AND THEIR ROLE
IN CERVICAL CANCER
 Antioxidant enzyme are a critical components of our body’s defense
system against oxidative stress , which can cause damage to cells &
DNA.
 These enzymes work by neutralizing harmful molecules called free
radicals, which are produced during metabolic processes as well as
exposure to environmental toxins like cigrette smoke and pollution.
 These are –
 SOD
 CAT
 GSH
 SOD (Superoxide dismutase): Converts superoxide radicals
into hydrogen peroxide. Superoxide is produced as a by-
product of oxygen metabolism and, if not regulated,
causes cell damage.
 CAT (Catalase): Break down hydrogen peroxide into water
and oxygen. In cervical cancer, CAT activity tends to decrease,
potentially leading to higher levels of hydrogen peroxide,
which can promote DNA damage and cancer development.
 GSH (Glutathione): GSH is the most abundant non-protein
thiol present at millimolar concentrations in mammalian
tissues. As an important intracellular antioxidant, it acts as a
regulator of cellular redox state protecting cells from damage
caused by lipid peroxides, reactive oxygen and nitrogen
species, and xenobiotics.
OBJECTIVE
 Estimation of antioxidant enzymes superoxide
dismutase (SOD), catalase (CAT), and GSH
levels, in serum samples of cervical cancer
patients.
STUDY DESIGN
The inclusion and exclusion criteria for cases and
controls were as follows:
Inclusion criteria for cervical cancer patients
• Histopathologically proven cases of squamous cell
carcinoma (SCC), all stages and adenocarcinomas (AD).
• Women between 35-70 years with cervical cancer
symptoms such as vaginal discharge, pain in lower
abdomen and contact bleeding.
• Cervical biopsy positive.
 Exclusion Criteria for Cervical Cancer cases
• Women 70 years.
• Cases having double malignancy.
• Cases having any co-morbid condition such as diabetes,
tuberculosis etc.
• Cervical biopsy negative
 Inclusion criteria for control subject
• Healthy age matched
• Histopathologically negative for squamous cell
carcinoma, all stages and cervical intraepithelial
neoplasia (CIN).
• No previous history of any type of cancer.
RESULTS
BSA standard curve for protein estimation
Standard curve for BSA (0.1 - 1 mg/ml) concentration and for calculation of protein in
serum sample. A standard curve of BSA is made using absorbance versus micrograms
of protein and determining amounts from the curve .
Figure. BSA standard curve Antioxidant enzyme assays
CAT (CATALASE ACTIVITY)
Figure: Graphical representation of activities of catalase enzyme in
healthy subjects (controls) and cervical cancer cases.
SOD (SUPEROXIDE DISMUTASE)
Figure.: Graphical representation of activities of SOD enzyme in healthy
subjects (controls) and cervical cancer cases.
GSH ( GLUTATHIONE)
Figure.: Graphical representation of activities of GSH enzyme in healthy
subjects (controls) and cervical cancer cases.
DISCUSSION
 The evaluation of antioxidant enzymes viz. Superoxide
dismutase (SOD), Catalase (CAT) and Glutathione (GSH)
was carried out in 13normal control subjects and 50cervical
cancer cases
 There was significant increase in CAT activity in cervical
cancer cases .
 Significant decrease in GSH content and SOD activities in
cervical cancer cases was observed as compared to controls.
CONCLUSION
 We found statistically significant decrease in the SOD
activity and GSH and there was significant increase
in CAT activity in cervical cancer , when compared to
controls. This means that the antioxidant enzyme
production is affected in cervical cancer subjects leading
to higher risk of cell/organ damage. Therefore, detection
of these antioxidant enzyme levels can be used as
markers for monitoring cervical cancer in populations.
REFERENCES
 Halliwell, Barry (2007). "Oxidative stress and cancer: have we moved
forward?"
 Joe M. Mccord, irwin fridovich.(1969).Superoxide dismutase: AN
ENZYMIC FUNCTION FOR ERYTHROCUPREIN (HEMOCUPREIN),
journal of biological chemistry, volume 244, issue 22, 1969, pages 6049-
6055, ISSN 0021-9258, doi.Org/10.1016/s0021-9258(18)63504-5.
 GLOBOCAN. 2012. Database. Cervical cancer: estimated incidence,
mortality and prevalence worldwide in 2012. International Agency for
Research on Cancer. Lyon, France, [cited 2018 Jul 15].
 Cohen, P. A., Jhingran, A., Oaknin, A., & Denny, L. (2019). Cervical
cancer. Lancet (London, England), 393(10167), 169–182.
https://doi.org/10.1016/S0140-6736(18)32470-X
THANK YOU
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Oxidative stress and its implications in cervical cancer.pptx

  • 1. “OXIDATIVE STRESS AND ITS IMPLICATION IN CERVICAL CANCER” Submitted By Mahima Gupta Class: M.Sc. Biotechnology, IV Semester Roll No.: 2110014155034 Under the supervision of Prof. Monisha Banerjee Institute of Advanced Molecular Genetics & Infectious Diseases (IAMGID) ONGC Centre for Advanced Studies (OCAS) University of Lucknow Submitted to Department of Biochemistry, University of Lucknow
  • 2. CONTENTS  Introduction to cervical cancer  Oxidative Stress and Cervical Cancer  Antioxidant enzyme and their role in cervical cancer  Methodology Protein estimation GSH estimation Catalase estimation SOD Activity  Results  Discussion  Conclusion  References
  • 3. INTRODUCTION Cervical cancer is the third most common cancer among women in world. In India , cervical cancer is the second most common. Cervical cancer is a type of cancer that occurs in the cells of the cervix — the lower part of the uterus that connects to the vagina. Major cause is Human papillomavirus (HPV).
  • 4. CELL TYPES OF CERVICAL CANCER  Squamous cell :This type of cervical cancer begins in the thin, flat cells (squamous cells) lining the outer part of the cervix, which projects into the vagina. Most cervical cancers are squamous cell carcinoma.  Adenocarcinoma:This type of cervical cancer begins in the column-shaped glandular cells that line the cervical canal.
  • 5. According to Globocan 2020 total cases of cervical cancer were 1393717 in worldwide ( age group 5-74 years) among those 297119 were died.  In India estimated number of prevalent cases (5 years) in 2020, females ages 5-74 is 1654540 and total estimated deaths in 2020 females ages 5-74 is 363844. Prevalence of cervical cancer
  • 7. OXIDATIVE STRESS AND CERVICAL CANCER  Oxidative stress is an imbalance between the production of reactive oxygen species (ROS) and the body’s ability to detoxify them.  ROS can cause damage to cellular components such as DNA, proteins, and lipids, leading to cellular dysfunction and death.  This process has been implicated in the development and progression of various cancer, including cervical cancer.
  • 8. ANTIOXIDANT ENZYME AND THEIR ROLE IN CERVICAL CANCER  Antioxidant enzyme are a critical components of our body’s defense system against oxidative stress , which can cause damage to cells & DNA.  These enzymes work by neutralizing harmful molecules called free radicals, which are produced during metabolic processes as well as exposure to environmental toxins like cigrette smoke and pollution.  These are –  SOD  CAT  GSH
  • 9.  SOD (Superoxide dismutase): Converts superoxide radicals into hydrogen peroxide. Superoxide is produced as a by- product of oxygen metabolism and, if not regulated, causes cell damage.  CAT (Catalase): Break down hydrogen peroxide into water and oxygen. In cervical cancer, CAT activity tends to decrease, potentially leading to higher levels of hydrogen peroxide, which can promote DNA damage and cancer development.  GSH (Glutathione): GSH is the most abundant non-protein thiol present at millimolar concentrations in mammalian tissues. As an important intracellular antioxidant, it acts as a regulator of cellular redox state protecting cells from damage caused by lipid peroxides, reactive oxygen and nitrogen species, and xenobiotics.
  • 10. OBJECTIVE  Estimation of antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and GSH levels, in serum samples of cervical cancer patients.
  • 12. The inclusion and exclusion criteria for cases and controls were as follows: Inclusion criteria for cervical cancer patients • Histopathologically proven cases of squamous cell carcinoma (SCC), all stages and adenocarcinomas (AD). • Women between 35-70 years with cervical cancer symptoms such as vaginal discharge, pain in lower abdomen and contact bleeding. • Cervical biopsy positive.  Exclusion Criteria for Cervical Cancer cases • Women 70 years. • Cases having double malignancy. • Cases having any co-morbid condition such as diabetes, tuberculosis etc. • Cervical biopsy negative
  • 13.  Inclusion criteria for control subject • Healthy age matched • Histopathologically negative for squamous cell carcinoma, all stages and cervical intraepithelial neoplasia (CIN). • No previous history of any type of cancer.
  • 14. RESULTS BSA standard curve for protein estimation Standard curve for BSA (0.1 - 1 mg/ml) concentration and for calculation of protein in serum sample. A standard curve of BSA is made using absorbance versus micrograms of protein and determining amounts from the curve . Figure. BSA standard curve Antioxidant enzyme assays
  • 15. CAT (CATALASE ACTIVITY) Figure: Graphical representation of activities of catalase enzyme in healthy subjects (controls) and cervical cancer cases.
  • 16. SOD (SUPEROXIDE DISMUTASE) Figure.: Graphical representation of activities of SOD enzyme in healthy subjects (controls) and cervical cancer cases.
  • 17. GSH ( GLUTATHIONE) Figure.: Graphical representation of activities of GSH enzyme in healthy subjects (controls) and cervical cancer cases.
  • 18. DISCUSSION  The evaluation of antioxidant enzymes viz. Superoxide dismutase (SOD), Catalase (CAT) and Glutathione (GSH) was carried out in 13normal control subjects and 50cervical cancer cases  There was significant increase in CAT activity in cervical cancer cases .  Significant decrease in GSH content and SOD activities in cervical cancer cases was observed as compared to controls.
  • 19. CONCLUSION  We found statistically significant decrease in the SOD activity and GSH and there was significant increase in CAT activity in cervical cancer , when compared to controls. This means that the antioxidant enzyme production is affected in cervical cancer subjects leading to higher risk of cell/organ damage. Therefore, detection of these antioxidant enzyme levels can be used as markers for monitoring cervical cancer in populations.
  • 20. REFERENCES  Halliwell, Barry (2007). "Oxidative stress and cancer: have we moved forward?"  Joe M. Mccord, irwin fridovich.(1969).Superoxide dismutase: AN ENZYMIC FUNCTION FOR ERYTHROCUPREIN (HEMOCUPREIN), journal of biological chemistry, volume 244, issue 22, 1969, pages 6049- 6055, ISSN 0021-9258, doi.Org/10.1016/s0021-9258(18)63504-5.  GLOBOCAN. 2012. Database. Cervical cancer: estimated incidence, mortality and prevalence worldwide in 2012. International Agency for Research on Cancer. Lyon, France, [cited 2018 Jul 15].  Cohen, P. A., Jhingran, A., Oaknin, A., & Denny, L. (2019). Cervical cancer. Lancet (London, England), 393(10167), 169–182. https://doi.org/10.1016/S0140-6736(18)32470-X