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. 2021 Jan-Mar;13(1):42-46.
doi: 10.18502/ajmb.v13i1.4577.

β-sitosterol Mediated Silver Nanoparticles Induce Cytotoxicity in Human Colon Cancer HT-29 Cells

Palaniappan Chithambara Shathviha  1 , Devaraj Ezhilarasan  1 , Shanmugam Rajeshkumar  1 , Jayaraman Selvaraj  2
Affiliations

β-sitosterol Mediated Silver Nanoparticles Induce Cytotoxicity in Human Colon Cancer HT-29 Cells

Palaniappan Chithambara Shathviha et al. Avicenna J Med Biotechnol. 2021 Jan-Mar.

Abstract

Background: Silver nanoparticles (AgNP) are commonly used metallic nanoparticles in health care systems. Colon cancer incidence is increasing worldwide. In this study, AgNP was synthesized using β-sitosterol and its cytotoxic potential was evaluated in human colon cancer (HT-29) cells.

Methods: Characterization of AgNP was analyzed by TEM and spectrophotometry analysis. HT-29 cells were treated with different concentrations (2, 4, 6, 8 and 10 ng/ml) of AgNPs and cytotoxicity was evaluated by MTT assay. The apoptosis was analyzed by the flow cytometry. The expression of p53 protein was analyzed by western blotting.

Results: β-sitosterol mediated AgNP are spherical in shape and induced concentration-dependent cytotoxicity in HT-29 cells. AgNP caused apoptosis related morphological changes as evidenced by annexin positive staining. AgNP treatments also induced the p53 expression in HT-29 cells.

Conclusion: Our present result suggests that β-sitosterol mediated AgNP induce apoptosis in colon cancer cells and this finding may pave the way for further experimental analysis in vivo.

Keywords: Apoptosis; Colon neoplasms; HT29 cells; Silver nanoparticles; β-sitosterol.

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Conflict of interest statement

Conflict of Interest The authors declare they have no conflict of interest.

Figures

Figure 1.
Figure 1.
Synthesis and characterization of AgNP using sitosterol. A) UV-vis spectroscopic spectrum of AgNP. B) TEM image of AgNP. C) Histogram shows the size distribution of nanoparticles. ImageJ software was used to measure different sizes of nanoparticles.
Figure 2.
Figure 2.
β-sitosterol mediated silver nanoparticles (AgNP) induced changes in the proliferation of HT-29 cells. A) Morphology of control and AgNP treated HT-29 cells, scale bar of 100 μm. B) Cytotoxi-city analysis by MTT assay, n=3, *** p<0.001 vs. control.
Supplementary file.
Supplementary file.
β-sitosterol Mediated Silver Nanoparticles synthesis and colour transition.
Figure 3.
Figure 3.
Cell viability analysis by calcein AM staining. Red arrow showing green fluorescence indicates the viable cells.
Figure 4.
Figure 4.
A) Morphological analysis of apoptosis by annexin V staining, B) Quantification of apoptotic cells, n=3, *** p<0.001 vs. control.
Figure 5.
Figure 5.
Flow cytometry analysis of apoptosis using annexin v and propidium iodide staining. Left upper quadrant percentage indicates the presence of apoptotic cells.
Figure 6.
Figure 6.
A) Western blot analysis of p53 expression. B) Quantification of p53 protein expression by densitometry analysis, n=3, *p< 0.05 and ** p<0.01 vs. control.
See this image and copyright information in PMC

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