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. 2019 Jan 25;20(1):123-130.
doi: 10.31557/APJCP.2019.20.1.123.

Evaluating of Induction of Apoptosis by Cornus Mass L. Extract in the Gastric Carcinoma Cell Line (AGS)

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Free PMC article

Evaluating of Induction of Apoptosis by Cornus Mass L. Extract in the Gastric Carcinoma Cell Line (AGS)

Farzaneh Sadat Hosseini et al. Asian Pac J Cancer Prev. .
Free PMC article

Abstract

Aim and objectives: Natural products and derivatives of medicinal vegetation can play an important role to the cure tumor. The Present study was focused to determine the effect of Cornus mass L. extract on the induction of apoptosis in AGS gastric carcinoma cell line in compared to L929 cells. Methods: In this experimental study, AGS and L929 cells were cultured and treated with different concentrations (0–10 mg/ml) of Cornus mass L. extract for 48 and 72 hours. Cell proliferation was assessed by MTT assay. The optical density of the colored solution was quantified at 570 nm wavelengths by an ELISA Reader. Making use of the apoptosis detection kit of Annexin V-FITC, PI and double staining with Annexin V-FITC were carried out for flow cytometry investigations. Data were analyzed by ANOVA. Variations with a P-value less than 0.05 were considered significant. Results: shows a noticeable deviation among various concentrations of extract when cells were treated for 48, 72 h declined cell viability in AGS cell line in comparison L929 cell lines in a dose and time-dependent manner (P < 0.05). This extract also displayed approximately several-fold increased anti-cancer potency in AGS compared to L929 cells. The IC50 value in AGS cells (evaluated after 48,72h) of the extract against AGS cells was 5/44, 2/44 mg/ml (p≤0.05). The analysis results of flow cytometry indicated that apoptosis was induced by the extract in AGS cells treated, compared with L929 cells. Conclusion: Each of our results implicates the reality that Cornus mass L. extract acts as a novel, potent inhibitor of cancer proliferation in in vitro. This may result in developing a promising therapeutic agent for the treatment of indole-sensitive cancers.

Keywords: Apoptosis; Gastric cancer; Cornus mass L. extract; L929 cells; AGS cell line.

Figures

Figure 1
Figure 1
The Effects of Cornus mass L. on AGS Cells’ Viability. The cells were exposed to various con¬centrations of Cornus mass L. in 48 hours and 72 hours, with the survival rates of the cells figured out by the MTT test. Every point of data yields a result average for three separate tests performed three times and reported as M ± SD.
Figure 2
Figure 2
Effects of Cornus mass L. on L929 Cells’ Viability. The cells were exposed to various concentrations of Cornus mass L. in 48 hours and 72 hours, with the survival rates of the cells figured out using the MTT test. Every point of data yields a result average for three separate tests performed three times and reported as M ± SD.
Figure 3
Figure 3
Morphological Changes on AGS(a-b) (control AGS (a), IC50AGS (b)) and L929 cells (c-d) (control L929 (c), IC50 L929 (d)) after exposure with Cornus mass L extract at 0 (untreated) and IC50 concentrations, respectively, that were observed with an inverted microscope.
Figure 4
Figure 4
Inducing Apoptosis in AGS Cells Making Use of Cornus mass L. extract. Cells were exposed to IC50 con¬centrations of Cornus mass L. extract for 48 hours and 72 hours, being smeared with propidium iodide (PI) and Annexin V fluorescein isothiocyanate (FITC). Consequently, flow cytometry was utilized to quantify necrotic and apoptotic cells. Various subsidiary populations were determined as Q1, i.e. PI positive and Annexin V negative, i.e. necrotic cells; Q2, i.e. Annexin positive V/PI double, i.e. late cells of apoptosis; Q3, Annexin negative V/PI double, i.e. ordinary living cells; and Q4, Annexin PI negative yet V positive, i.e. initial cells of apoptosis. *P<0.05 indicates a noticeable deviation from the control cells.
Figure 5
Figure 5
Inducing Apoptosis in L929 Cells Making Use of the Cornus mass L. extract. Cells were exposed to IC50 con-centrations of Cornus mass L. extract for 48 hours and 72 hours and smeared with propidium iodide (PI) and Annexin V fluorescein isothiocyanate (FITC). Next, flow cytometry was utilized to quantify necrotic and apoptotic cells. Various subsidiary populations were determined as Q1, PI positive but Annexin V negative, or necrotic cells; Q2, Annexin positive V/PI double, or late cells of apoptosis; Q3, Annexin V negative /PI double, or ordinary living cells; and Q4, PI negative but Annexin V positive, or initial cells of apoptosis. *P<0.05 shows a noticeable deviation from the control cells.
Figure 6
Figure 6
a, Indicates UL(necrotic cells), UR (late apoptotic cells), LL (normal live cells), LR( early apoptotic cells) in AGS cells after treatment with IC50 concentrations of Cornus mass L in comparison with untreated cells after 48,72 h; b, Indicates UL(necrotic cells), UR (late apoptotic cells), LL (normal live cells), LR( early apoptotic cells) in L929 cells after treatment with IC50 concentrations of Cornus mass L in comparison with untreated cells after 48,72 h.

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