Inhibition of Proliferation of HeLa Cells by Pulsed Electric Field Treated Mentha piperita (Mint) Extract

Abstract

In this research, the effectiveness of Mentha piperita (M. piperita) leaf extract against HeLa, a human cervical cancer cell line was investigated. We assessed the phytochemicals present in this extract by phytochemical screening tests and analysed the free radical scavenging activity and anticarcinogenic activity using without Pulsed Electric Field (PEF). The methanolic M. piperita extract was prepared by the Soxhlet extraction method, and was treated by PEF using 50 unipolar pulses, 10 kV/cm, 100 µs and the effect of PEF on the antioxidant and anticarcinogenic activities of M. piperita extract was investigated and compared to Soxhlet extraction (70°C for 8 h). The preliminary phytochemical analysis indicated the presence of phenols, tannins, flavonoids, alkaloids, steroids, reducing sugars and the absence of saponins, anthocyanins, catechins, terpenoids, carotenoids, and anthraquinones. The free radical scavenging activity of both PEF treated and untreated extract was determined by 2,2-Diphenyl-1-Picrylhydrazyl assay. Compared to untreated extract, 7.36% greater antioxidant activity was obtained from PEF treatment. The anticarcinogenic activity of both extracts were determined by MTT proliferation assay against Hela cells. The results indicated a viability of 34.77% for the PEF treated M. piperita extract (compared to 41.2% for the Soxhlet extract and 100% for the control). This indicates that the PEF-treated M. piperita extract showed higher antioxidant activity and anticarcinogenic activity than the other extracts. Hence, the application of PEF may be appropriate to achieve antioxidant rich extract and maximal cell death could be achieved with PEF treatment thereby, reducing the concentration of extract required for treating cancer.

KEYWORDS:

• Anticancer drug
• Antioxidant
• Cervical cancer
• HeLa cells
• Mentha piperita
• Organic leaf extracts
• Pulsed electric field

Additional information

Notes on contributors

Thulasidas Jeya Shree

Thulasidas Jeya Shree received the BE degree in electrical and electronics engineering from PSR. Engineering College, Sivakasi, India, in 2014 and ME degree from the College of Engineering, Guindy, Anna University, Chennai, India in 2016. She is a research student in the Division of High Voltage Engineering, College of Engineering, Anna University, Chennai, India. At present, she is a visiting researcher at Purdue University, West Lafayette, IN, USA. Her research interests include the application of pulsed electric field to enhance the extraction of bioactive compounds and to study the synergic effect of bioactive compound and pulsed electric field in treating cancer. E-mail: [email protected], [email protected]

Varadarajan Gowri Sree

Varadarajan Gowri Sree received her doctorate in High Voltage Engineering in 2008 from the College of Engineering, Guindy, Anna University, India. She is presently an associate professor in the same university with over 25 years of teaching experience. She has contributed a chapter to the book, titled, “Electroporation-based Therapies for Cancer: From Basics to Clinical Applications” (Wood head Publishing, 2014). Her current research topics are application of high voltage engineering for cancer treatment and enhanced bioactive compound extraction. E-mail: [email protected]

Sadasivam Poompavai

Sadasivam Poompavai received the BE degree in electrical and electronics engineering from Prince Shri Venkateshwara Padmavathy Engineering College, Anna University, in 2013. She received the ME degree in high voltage engineering from the College of Engineering, Guindy, Anna University, in 2015, and at present she is a research student in the Division of High Voltage Engineering, Anna University, Chennai, India. Her research interests include the application of pulsed electric fields to cancer, modelling of breast cancer, study of effectiveness of natural herbal sources as an alternative chemotherapeutic drug. E-mail: [email protected], [email protected]

Elisabetta Sieni

Elisabetta Sieni received her degree in electronic engineering in 2002 and her Doctorate in bio-electromagnetics and electromagnetic compatibility and in information engineering, bioengineering, from Padova University in 2006 and 2011, respectively. She is currently working at the Department of Industrial Engineering of Padova University as a post-doc. Her research interest is focused on biomedical application of electromagnetic fields. E-mail: [email protected]

Paolo Sgarbossa

Paolo Sgarbossa graduated in chemistry at the University of Padua in 2003 discussing a thesis on the “Synthesis, coordination to transition metals and reactivity of N-functionalized isocyanides” and the PhD degree in chemical science at the same University in 2007 with a thesis on “Platinum (II) complexes as Lewis Acids in catalytic oxidation reactions with hydrogen peroxide”. He is currently associate professor at the Department of Industrial Engineering at the University of Padova. E-mail: [email protected]

Ignacio Camarillo

Ignacio Camarillo is an associate professor in the Department of Biological Sciences at Purdue University. He received his PhD in the Department of Physiology from Wayne State University. His work was supported by the Minority Biomedical Research Support (MBRS) Program. He continued his training as a National Cancer Institute Postdoctoral Fellow in the Department of Molecular Cellular and Developmental Biology at the University of California Santa Cruz, where he studied “ The protective effect of pregnancy against breast cancer”. He is a member of the Purdue Center for Cancer Research and serves on the Executive Committee for the Obesity and Cancer Discovery group. His current research focuses on the mechanisms involved in the relationship between diet, obesity, and breast cancer. E-mail: [email protected]

Raji Sundararajan

Raji Sundararajan is a professor at the School of Engineering Technology, Purdue University. She obtained her initial training on electroporation at Johns Hopkins Medical School and has been doing research on electroporation for the past several years. She was the recipient of the 2010 Indiana ‘Women & Hi-Tech’ award for ‘Distinguished use of technology in healthcare Life Science’ and won a prestigious, 1-year (2010/11) fellowship to study regenerative medicine therapy using optoinjection and electroporation techniques at Purdue. She is a reviewer of NIH, NSF, US International Science & Technology Center, and US National Research Council proposals, and various scholarly journals including International Journal of Cancer, Molecular Biotechnology, Journal of Biomedical Microdevices, Journal of Anticancer Drugs, and several IEEE Transactions.