Evaluation of cytotoxicity of aqueous extract of Graviola leaves on squamous cell carcinoma cell-25 cell lines by 3-(4,5-dimethylthiazol-2-Yl) -2,5-diphenyltetrazolium bromide assay and determination of percentage of cell inhibition at G2M phase of cell cycle by flow cytometry: An in vitro study

Visveswaraiah Paranjyothi Magadi, Venkatadasappa Ravi, Anantharaju Arpitha, Litha, Kikkerilakshminarayana Kumaraswamy, Krishnappa Manjunath, Visveswaraiah Paranjyothi Magadi, Venkatadasappa Ravi, Anantharaju Arpitha, Litha, Kikkerilakshminarayana Kumaraswamy, Krishnappa Manjunath

Abstract

Introduction: Malignancies constitute a wide variety of disorders having high mortality and morbidity rates. Current protocols for management include surgical intervention, chemotherapy, and radiation which possess numerous adverse effects. Many phytochemicals are available with anticancer properties similar to anticancer drugs. Major benefit of these compounds is apparent lack of toxicity to normal tissues. Graviola (botanical name: Annona Muricata) contain bioactive compound "annonaceous acetogenins" known for anticancer activity on cancer cell lines.

Aims: To determine cytotoxicity of Graviola and percentage cell inhibition at G2M phase of cell cycle.

Settings and design: The cytotoxicity of aqueous extract of Graviola leaves on squamous cell carcinoma (SCC-25) cell lines at various concentrations evaluated using 3-(4,5-dimethylthiazol-2-Yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The percentage of SCC-25 cell inhibition at G2M phase of cell cycle determined using flow cytometry.

Methods: Graviola Leaves, American Type Culture Collection SCC-25 cell lines were procured from Skanda Laboratories, Bengaluru. The cytotoxicity of aqueous extract of Graviola on SCC-25 cells at various concentrations evaluated using MTT assay. The percentage of SCC-25 cell inhibition at G2M phase of cell cycle determined using flow cytometry.

Statistical analysis: Statistical analysis was done using one-way ANOVA.

Results: MTT assay showed statistically significant (P < 0.001) dose-dependent inhibition of SCC-25 cell lines by Graviola with IC50 value of 12.42 μg/ml. Flow cytometry revealed that Graviola at 25 and 50 g/ml arrested 53.39% and 52.09% cells in G2M phase of cell cycle respectively, which was statistically significant.

Conclusion: Graviola showed significant cytotoxic activity and percentage of cell inhibition at G2M phase cell cycle against SCC-25 cell lines.

Keywords: 3-(4,5-Dimethylthiazol-2-Yl)-2,5-diphenyltetrazolium bromide assay; G2M phase; Graviola; flow cytometry; squamous cell carcioma-25 cell lines.

Figures

Graph 1
Graph 1
Aqueous extract of Graviola showed dose-dependent percentage inhibition of squamous cell carcinoma-25 cells
Figure 1
Figure 1
Histogram representation of flow cytometric analysis of the aqueous extract of Graviola at 25 μg/ml concentration on squamous cell carcinoma-25 cells
Figure 2
Figure 2
Histogram representation of flow cytometric analysis of control (untreated cells)
Figure 3
Figure 3
Histogram representation of flow cytometric analysis of colchicine on squamous cell carcinoma-25 cells
Figure 4
Figure 4
Histogram representation of flow cytometric analysis of the aqueous extract of Graviola at 50 μg/ml concentration on squamous cell carcinoma-25 cells

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Source: PubMed

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