ABSTRACT
Objective:To determine the effect of rice bran extract (RBE) in combination with doxorubicin on 4T1 triple-negative breast cancer cells and NIH-3T3 cells. Methods:RBE was obtained by maceration with n-hexane. The phytochemical profile of RBE was observed using high-performance liquid chromatography. Cytotoxic effect of RBE was evaluated through MTT assay. In addition, flow cytometry was used for cell cycle and apoptosis analysis. Cellular senescence was observed using SA-β-Gal assay and intracellular reactive oxygen species (ROS) levels were evaluated using DCFDA staining. The pro-oxidant property of RBE was also evaluated through 1-chloro-2,4-dinitrobenzene spectrophotometry and molecular docking. Results:RBE was obtained with a yield of 18.42％ w/w and contained tocotrienols as the major compound. RBE exerted no cytotoxic effect on 4T1 and NIH-3T3 cells. However, RBE in combination with doxorubicin decreased 4T1 cell viability synergistically (combination index<0.9) and induced apoptosis and senescence on 4T1 cells. RBE significantly decreased senescence in doxorubicin-treated NIH-3T3 cells. Additionally, RBE did not increase ROS levels in doxorubicin-treated 4T1 cells. Meanwhile, the combination of RBE and doxorubicin reduced ROS levels in NIH-3T3 cells. RBE significantly reduced glutathione-S-transferase activity and alpha-tocotrienol interacted with glutathione-S-transferase in the glutathione binding site. Conclusions:Rice bran may be used as a co-chemotherapeutic agent to improve the therapeutic effectiveness of doxorubicin while protecting against the cellular senescence effects of doxorubicin on healthy cells.
ABSTRACT
Long-term use of doxorubicin (DOX) causes several side effects, especially induction of metastasis, in breast cancercells. Pentagamaboronon-0 (PGB-0) or 2,5-bis(4-boronic acid benzylidene) cyclopentanone is a novel curcumin analogthat exerts cytotoxic effects on Human Epidermal Growth Factor Receptor 2 (HER2)-positive breast cancer cells. Theobjective of this study was to evaluate PGB-0 as a co-chemotherapeutic agent on DOX-induced metastatic breastcancer cells, 4T1. Potential cytotoxic and antimetastatic activities of PGB-0 were screened by molecular docking underPLANTS software, which revealed that the PGB-0 interacted with matrix metalloproteinase-9 (MMP-9) and InhibitorKappa β Kinase (IKKβ). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay showed that PGB-0 andDOX exhibited cytotoxic effects on 4T1 breast cancer cells, with IC50 values of 294 and 2.4 µM, respectively, andsynergistically increased the cytotoxicity of DOX. Results of propidium iodide staining flow cytometry revealed thatthe PGB-0 and its combination with DOX induced cell cycle arrest in the S phase and the G2/M phase, respectively.In addition, PGB-0 and its combination with DOX induced apoptosis. Regarding the antimetastatic activity, a singletreatment with PGB-0 inhibited cell migration, while its combination with DOX inhibited cell migration with morepotency than that with single treatment, as assessed through wound healing assay. Gelatin zymography revealed thatthe PGB-0 and its combination with DOX inhibited MMP-9 activity. Immunoblotting assay showed that the PGB-0and its combination with DOX decreased the expression of Rac1 and p120. In conclusion, PGB-0 increased thecytotoxicity and inhibited the induction of metastasis by DOX in breast cancer cells.
ABSTRACT
Cisplatin is drug of choice toward cervical cancer despite having many side effects, thus researches are conducted in order to find the effective and synergistic co-chemoterapeutic agent combined with cisplatin. In this study, we observed the potential of the cinnamon essential oil (CEO) isolated from Cinnamomum burmannii as cochemoterapeutic agent of cisplatin on HeLa cells covering cytotoxic effect, cell cycle modulation and induction of apoptosis. Cytotoxic effect was determined by using MTT assay; while induction of apoptosis and cell cycle profile were observed by using flow cytometry. At 24 hours of incubation, CEO showed cytotoxic effect on HeLa cells with IC50 value of 250 μg/mL, while cisplatin showed cytotoxic effect with IC50 value of 18 μM. Combination of CEO and cisplatin reduced cells viability compared to cisplatin solely. Moreover, flow cytometry using annexin-V and PI showed that CEO and its combination with cisplatin induced apoptosis lower than cisplatin alone at 24 hours of incubation. Further analysis on the cell cycle progression showed that CEO induced S-phase arrest on HeLa cells, cisplatin induced G1 arrest, while combination of CEO and cisplatin induced G2/M arrest. Thus, the inhibition of HeLa cells growth at 24 hours is likely through cell cycle modulation rather than apoptosis.
ABSTRACT
The cytotoxic activity and apoptosis induction of 8-hydroxyisocapnolactone-2΄,3΄-diol (HICD) and its combination with doxorubicin (Doxo) on MCF-7 and T47D cells have been evaluated. The cytotoxic assay was performed using MTT method. The IC50 was used to express the cytotoxic potency, while the combination index (CI) was calculated to determine the effect of combination. The apoptotic assay was carried out using acrydine orange - ethidium bromide double staining method, while Bcl-2 and Bax expression were investigated by immunocytochemistry. The HICD exhibited cytotoxic activity on MCF-7 and T47D cells with the IC50 of 8 μg/ml (21.2 μM) and 4 μg/ml (10.6 μM), respectively. The combination of HICD with Doxo showed synergistic effect and increased apoptosis induction on both cell lines. The HICD did not affect Bcl-2 but increased Bax expression on MCF-7 cells, while on T47D cells it suppressed Bcl-2 expression but did not modulate Bax expression.
ABSTRACT
Objective:To evaluate of hesperidin to overcome resistance of doxorubicin in MCF-7 resistant doxorubicin cells (MCF-7/Dox) in cytotoxicity apoptosis and P-glycoprotein (Pgp) expression in combination with doxorubicin. Methods:The cytotoxic properties, 50%inhibition concentration (IC50) and its combination with doxorubicin in MCF-7 cell lines resistant to doxorubicin (MCF-7/Dox) cells were determined using MTT assay. Apoptosis induction was examined by double staining assay using ethidium bromide-acridine orange. Immunocytochemistry assay was performed to determine the level and localization of Pgp. Results: Single treatment of hesperidin showed cytotoxic activity on MCF-7/Dox cells with IC50 value of 11 μmol/L. Thus, combination treatment from hesperidin and doxorubicin showed addictive and antagonist effect (CI>1.0). Hesperidin did not increase the apoptotic induction, but decreased the Pgp expressions level when combined with doxorubicin in low concentration. Conclusions: Hesperidin has cytotoxic effect on MCF-7/Dox cells with IC50 of 11 μmol/L. Hesperidin did not increased the apoptotic induction combined with doxorubicin. Co-chemotherapy application of doxorubicin and hesperidin on MCF-7/Dox cells showed synergism effect through inhibition of Pgp expression.
ABSTRACT
The selaginella ethanolic extract shows cytotoxic activity against T47D and MCF-7 cells. The aim of this research is to evaluate the cytotoxic effect and apoptosis induction of selaginella fractions on MCF-7 cells. The Selaginella plana powder was extracted by absolute ethanol. Ethanolic extract was dilluted by methanol:water (4:1) and then fractionated by hexane (S_Hex), methylene chloride (S_MTC), ethyl acetate (S_EA), and buthanol (S_BuOH). Cytotoxic activity was examined by MTT assay. Apoptosis examination used acrydine orangeetidium bromide staining (double staining). The result showed that the IC50 value of S_Hex, S_MTC, S_EA, and S_BuOH on MCF-7 cells were 30 μg/mL, 19 μg/mL, 24 μg/mL, and 2 μg/mL respectively. The active fractions (S_Hex, S_MTC, S_EA and S_BuOH) at its IC50 concentration increased apoptotic cells on the MCF-7 cells 35.33%, 20.33%, 24% and 45.67% respectively compared to control. Based on the result, buthanol fraction of Selaginella plana (S_BuOH) showed the highest apoptotic induction on MCF-7 cancer cells.
ABSTRACT
Objective: To investigate the estrogenic effect of (8,9)-furanyl-pterocarpan-3-ol (FPC) on growth of human breast cancer T47D cells and the interactions between the FPC and tamoxifen (TAM), on the growth of estrogen receptor-dependent breast cancer T47D cells.Methods:T47D cells were treated with FPC alone (0.01-200 μmol/L) or in combination with TAM 20 nmol/L. The proliferation effect of FPC were conducted on T47D cells in vitro by MTT test. Furthermore, the expression of ERα or c-Myc were also determined by immunohistochemistry.Results:inhibitory effect on T47D cells, wheraes co-administered with low concentration (less than 1μmol/L) of FPC attenuated to promote cell proliferation. In contrast, the combination of TAM with higher doses (more than 20 μmol/L) of FPC showed growth inhibitory. This result was supported by immunocytochemistry studies that the administration of 20 nmol/L TAM down-regulated ER-αand c-Myc, but the combination of 20 nmol/L TAM and 1 μmol/L FPC robustly up-regulated expression of ER-α. Thus, the reduced growth inhibition of TAM 20 nmol/L by FPC 1 μmol/L on T47D cells may act via the modulation of ER-α.Conclusions:The findings indicate and suggest that FPC had estrogenic activity at low The results indicated that administration of an anti-estrogen TAM showed growth concentrations and anti-estrogenic effect that are likely to be regulated by c-Myc and estrogen receptors. We also confirm that low concentration of FPC attenuated the growth-inhibitory effects of TAM on mammary tumor prevention. Therefore, the present study suggests that caution is warranted regarding the consumption of dietary FPC by breast cancer patients while on TMA therapy.
ABSTRACT
Objective: To evaluate the effects of n-hexane insoluble fraction (HIF) of Ficus septica leaves in combination with doxorubicin on cytotoxicity, cell cycle and apoptosis induction of breast cancer T47D cell lines. Methods: The in vitro drugs-stimulated cytotoxic effects were determined using MTT assay. Analysis of cell cycle distribution was performed using flowcytometer and the data was analyzed using ModFit LT 3.0 program. Apoptosis assay was carried out by double staining method using ethydium bromide-acridin orange. The expression of cleaved-poly (ADP-ribose) polymerase (PARP) on T47D cell lines was identified using immunocytochemistry. Results:The combination exhibited higher inhibitory effect on cell growth than the single treatment of doxorubicin in T47D cells. In addition, combination of doxorubicin and HIF increased the incidence of cells undergoing apoptosis. HIF could improve doxorubicin cytotoxic effect by changing the accumulation of cell cycle phase from G2/M to G1 phase. The combination also exhibited upregulation of cleaved-PARP in T47D cells. Conclusions: Based on this results, HIF is potential to be developed as co-chemotherapeutic agent for breast cancer by inducing apoptosis and cell cycle arrest. However, the molecular mechanism need to be explored further.