Modulation of Cell-Cycle gene expression by Chitosan coated Silver nanopartices on Human Epidermoid Carcinoma cells.MODULATION OF CELL-CYCLE GENE EXPRESSION BY CHITOSAN COATED SILVER NANOPARTICES ON HUMAN EPIDERMOID CARCINOMA CELLS

  • Muppalaneni Pavani SASTRA University
  • Sundaramurthy Dhakshinamoorthy SASTRA University
  • Kuppuswamy Ashok Ayyappa Assistant Professor( Research) Department of Biotechnology, School of Chemical and Biotechnology, SASTRA University

Abstract

Objective: To detect the variability in the expression of cell cycle genes such as Cyclin D1,CDK4 p21,p27 and p57 associated with cytotoxicity of Human epidermoid carcinoma cells using qPCR.

Methods: The synthesis silver nanoparticles and chitosan coated silver nanoparticles was done by chemical reduction method using polyvinyalpyrolidone (PVP) as surfactant and glucose as reducing agent. The cell proliferation assay was performed using MTS assay. Gene expression analysis was performed using Eppendorf Realplex 2 PCR systems.

Results: The lowest concentration of 1mg of Chitosan-AgNPs with 25μg/ml concentration showed 35% reduction in the A431 cell line growth. Significantly increased expression of p21, p27, p57, CCND1 and CD4 gene was observed among 1mg of Chitosan-AgNPs with 25μg/ml concentrations indicating potential apoptotic activity at the lowest concentration of Chitosan-AgNPs on A431 cells.

Conclusions: The effect of AgNPs inducing apoptosis/growth arrest at low concentrations (1mg-25μg/ml) has been more effective while coating the nanoparticle with chitosan. The potential apoptotic properties of Chitosan-AgNPs are evident by the significant up regulation of the cell cycle genes when treated with low concentration of Chitosan-AgNPs.

 

Keywords: Silver nanoparticles, Cell Cycle genes, Apoptosis, Chitosan, Gene expression

Downloads

Download data is not yet available.

Author Biography

Kuppuswamy Ashok Ayyappa, Assistant Professor( Research) Department of Biotechnology, School of Chemical and Biotechnology, SASTRA University

Gained 5 years of research experience in Genetics of type 2 diabetes and dyslipidemia.  My research training as part of the PhD studies consisted of both clinical /population based biochemical studies and Molecular genetic studies involving screening of Novel genetic variants associated with low HDL and High Triglyceride levels among diabetic and non-diabetic subjects.  I have screened over 25 SNPs spanning 5 candidate genes implicated in dyslipidemia and type 2 diabetes.  Determination of haplotypes that could be used as predictive tool in the inheritance of risk was also assessed in my research pertaining to low HDL risk and High Triglyceride risk.

My Current research work focuses on functional validation of some of the novel loci identified in the Genome Wide association studies (GWAS) linked to cardiovascular disease (CVD) using animal models, and regulation of these loci by specific miRNA’s.

 

 

Assistant Professor (Research),

Department of Biotechnology

References

1. Rasmus Foldbjerg, Day Anh Dang, Herman Autrup. Cytotoxicity and genotoxicity of silver nanoparticles in human lung cancer cell lines. Arch Toxicol 2011;85:743–50.
2. OV Salata. Applications of nanoparticles in biology and medicine. J Nanobiotechnol 2004;1477-3155.
3. J Saraniya Devi, B Valentin Bhimba. Silver nanoparticles: antibacterial activity against wound isolates and invitro cytotoxicity on human caucasion colon adenocarcinoma. Asian Specific J Trop Dis 2012;2(12):S87-S93.
4. MA Hettiarachchi, PASR Wickaramachchi. Synthesis of chitosan stabilized silver nanoparticles using gamma ray irradiation and characterization. J Sci Univ Kelaniya 2011;6:65-75.
5. J Saraniya Devi, B Valentin Bhimba. Anticancer activity of silver nanoparticles synthesized by the seaweed ulva lactuca In vitro studies. Open Access Sci Reports 2012;1:4.
6. Muthu Irulappan Sriram, Selvaraj Barath Mani Kanth, Kalimuthu Kalishwaralal Sangiliyandi gurunathan. Antitumor activity of silver nanoparticles in Dalton’s lymphoma ascites tumor model. Int J Nanomed 2010;5:753–62.
7. Dongwei Wei, Wuyong Sun, Weiping Qian, Yongzhong Ye, Xiaoyuan Ma. The synthesis of chitosan-based silver nanoparticles and their antibacterial activity. Carbohydr Res 2009;34(17):2375–82.
8. Shilpa Sharma, Pallab Sanpui, Arun Chattopadhyay, Siddhartha Sankar Ghosh. Fabrication of antibacterial silver nanoparticle—sodium alginate–chitosan composite films. RSC Adv 2012;2:5837–43.
9. KSV Krishna Rao, P Ramasubba Reddy, Yong-Ill Lee, Changdae Kim. Synthesis and characterization of chitosan–PEG–Ag nanocomposites for antimicrobial application. Carbohydr Polym 2012;87(1):920–5.
10. Monica Potara, Monica Baia, Cosmin Farcau, Simion Astilean. Chitosan-coated anisotropic silver nanoparticles as a SERS substrate for single-molecule detection. Nanotechnol 2012;23(5): 055501.
11. Sanpui P, Chattopadhyay A, Ghosh SS. Induction of apoptosis in cancer cells at low silver nanoparticle concentrations using chitosan nanocarrier. ACS Appl Mater Interfaces 2011;3(2):218-28.
12. Mocanu A, Pasca RD, Tomoaia G, Garbo C, Frangopol PT, Horovitz O. Tomoaia-Cotisel New procedure to synthesize silver nanoparticles andtheir interaction with local anesthetics. Int J Nanomed 2013;8:3867-74.
13. Chu IM, Hengst L, Slingerland JM. "The Cdk inhibitor p27 in human cancer: prognostic potential and relevance to anticancer therapy". Nat Rev Cancer 2008;8(4):253–67.
14. Matsuoka S, Edwards MC, Bai C, Parker S, Zhang P, Baldini A, et al. p57kip2, a structurally distinct member of the p21cip1 Cdk inhibitory family, is a candidate tumour suppressor gene. Genes Dev 1995;9:650–62.
15. Atsushi Tsugu, Keiichi Sakai, Peter B Dirks, Shin Jung, Rosanna Weksberg, Yan-Ling Fei, et al. Expression of p57KIP2 potently blocks the growth of human astrocytomas and induces cell senescence. Am J Pathol 2000;157(3):919-32.
16. Brady RL, Dodson EJ, Dodson GG, Lange G, Davis SJ, Williams AF, et al. (May 1993). "Crystal structure of domains 3 and 4 of rat CD4: relation to the NH2-terminal domains". Sci 1993;260(5110):979–83.
Statistics
272 Views | 1369 Downloads
How to Cite
Pavani, M., S. Dhakshinamoorthy, and K. A. Ayyappa. “Modulation of Cell-Cycle Gene Expression by Chitosan Coated Silver Nanopartices on Human Epidermoid Carcinoma cells.MODULATION OF CELL-CYCLE GENE EXPRESSION BY CHITOSAN COATED SILVER NANOPARTICES ON HUMAN EPIDERMOID CARCINOMA CELLS”. International Journal of Pharmacy and Pharmaceutical Sciences, Vol. 7, no. 2, Dec. 2014, pp. 355-9, https://innovareacademics.in/journals/index.php/ijpps/article/view/3698.
Section
Original Article(s)