Objective: This study intends to explore the potential of galangal extract as a co-chemotherapeutic agent through the analysis of its cytotoxic and migratory effects on metastatic breast cancer cells and as an anti-ageing agent through its senescence inhibitory effect on normal fibroblast cells. Methods: Galangal ethanolic extract (GE) was subjected to a cytotoxicity test with the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay alone or in combination with doxorubicin (Dox) against 4T1 cells but not in NIH-3T3 cells. Evidence of senescent cells was detected using a SA-β galactosidase based assay. In addition, the level of reactive oxygen species (ROS), apoptosis, and cell cycle were measured with a flow cytometry-based assay. Meanwhile, cell migration and matrix metalloproteinase (MMP)-9 expression after GE treatment on 4T1 cells were measured using the scratch wound healing assay and gelatin zymography assay, respectively. The metabolomic profiles of GE were traced using gas chromatography-mass spectrometry (GC-MS) analysis. Results: GE effectively inhibited the growth of 4T1 cells with an IC 50 value of 135 µg/mL and increased the cytotoxic effect of Dox at concentrations of 50 and 100 µg/mL. GE increased the number of senescent cells arrested in the G2/M phase but did not cause apoptosis. This effect is compounded by increasing intracellular levels of ROS. However, GE reduced senescence to normal in fibroblast cells (NIH 3T3 cells) under oxidative stress by Dox without any changes in the ROS level. Moreover, GE also inhibited the migration of 4T1 cells and suppressed the expression of MMP-9 induced by Dox. Conclusion: Galangal has the potential for use as a co-chemotherapeutic agent by inducing senescence in correlation with increasing intracellular ROS toward metastatic breast cancer. However, the effect of GE in decreasing the senescence phenomena toward normal fibroblast cells illustrates its potential as a promising anti-ageing agent.

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Objectives: This experiment aims to investigate the apoptosis effect of curcumin and its analogs pentagamavunon-0 (PGV-0) and PGV-1 on normal and other cancer cell lines.Methods: Growth inhibition effect was investigated using the MTT method. Double staining used acridine orange, 2-(4-aminodiphenyl)-6-indolcarbamidine dihydrochloride and ethidium bromide was performed to determine morphological changes of cells. Detection of PARP, caspase-3, PUMA and BAX using a western blot method was conducted to elucidate the apoptosis effect of the compounds.Results: PGV-1 (2.5 μM) and PGV-0 (5.0 μM) could inhibit T47D-cell growth on 72 h observation, but not for curcumin. DNA staining showed PGV-1 has the strongest apoptosis induction effect on T47D-cells compared to PGV-0 and curcumin as well. Western blot analysis resulted in cleavage PARP (83 kD) on HeLa, T47D, and MCF-7 cells treated with PGV-1 (2.5 µM), PGV-0 (5.0 µM). Curcumin (10.0 µM) just induced apoptosis on T47D-cell and MCF-7 cell, but not HeLa cell. Cleavage PARP resulted by apoptosis process in the cell. PGV-1 (2.5 µM) had a stronger apoptosis effect compared to PGV-0 (5.0 µM) and curcumin (10.0 µM) based on cleaved PARP result qualitatively. On the normal cell (NH3T3), cells that were treated with the compounds resulted in a negative cleavage PARP. This result indicated that the compounds were part of a selectively induced cancer cell line apoptosis process. Conclusion:Curcumin, PGV-0 and PGV-1 could inhibit cell growth by induce apoptosis on cancer cells but not on normal cells, which PGV-1 has strongest apoptosis induction effect on cancer cell lines.

Previous research stated that galangal (Alpinia galanga) extract has a potential as cytotoxic agent with active compound of 1’-Acetoxychavicol Acetate (ACA). The objective of this study was to determine the selectivity of ethanol extract, ethyl acetate fraction, and methanol fraction of of galangal, and ACA on cancer cell lines. Cytotoxic activity was carried out using the MTT method on T47D breast cancer, WiDr colon cancer, HeLa cervical cancer, and Vero normal cell lines. The results showed that galangal ethanol extract and its fractions had selectivity index equal to or less than 2 on cancer cells. Meanwhile, ACA had selectivity index more than 3 on T47D cell and HeLa cell. ACA showed a strong cytotoxic activity against cancer cells T47D, HeLa, and WiDr with IC50 values of 3.14, 7.26, and 12.49 μg/ml, respectively. Based on data, it could be concluded that ACA was the most selective to inhibit T47D cell with a selectivity index of 6.6.Keywords: 1’-Acetoxychavicol acetate, galangal (Alpinia galanga), selective index, cytotoxic

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Universitas Muhammadiyah Surakarta, Jawa Tengah, IndonesiaDue to severe side effect and non-specific chemotherapeutic agent, screening and discovery for cancer therapy are still working, especially from natural resources. Traditionally, people used herbal medicine either to prevent or cure diseases. One of herbal that commonly used in Indonesia is Alpinia galangal. Previous study stated that active compound is acetoxy chavicol acetate (ACA) and active as anticancer. This research aimed to determine ACA concentration and cytotoxic activity of Alpinia galanga extract (AGE) from three local markets on HeLa, MCF7 and T47D cell lines. The galangal used from three local markets namely Pasar Legi Surakarta, Beringharjo Yogyakarta, and Wonogiri. The extraction was performed by maceration using 96% ethanol as solvent. ACA quantitation using UV spectrophotometer at λ = 208.5 nm. Samples were prepared by liquid-liquid extraction using an ethyl acetate. Cytotoxic activities were performed by MTT assay. The result showed that the concentration of ACA of AGE from the three local markets were 3.798; 0.035; and 0.009 % w/w, respectively. Cytotoxic activity, describes as IC50 value, on HeLa cell line of AGE from three local markets, in order were 13.26; 36.32 and > 100 µg/ mL. Meanwhile, AGE from Pasar Legi on MCF7 and T47D cell lines have IC50 value of 15.80; 12.50 µg/ mL, respectively. In contrast, two other samples have IC50 values of greater than 100 µg/ mL. The highest activity was from the highest concentration of ACA on the samples.

Background: Previous research showed that the extract of guazuma leaves had cytotoxic activity on various cancer cell lines. One of the chemical markers of Guazuma ulmifolia Lamk. is tiliroside. Aims and Objectives: The aim and objective of this study was to determine the cytotoxic effects of tiliroside on T47D and MCF7 and their mechanisms. Materials and Methods: Tiliroside (was isolated by laboratory Biological Pharmacy of Universitas Muhammadiyah Surakarta, was confirmed by using liquid chromatography-tandem mass spectrometry/MS). The antiproliferative activity was analyzed by flow cytometry and immunocytochemistry methods. Results: Tiliroside has cytotoxic activity with inhibitory concentration 50% on T47D and MCF7 which was 67.79 and 100.00 µg/ml, respectively. Based on the immunocytochemistry, tiliroside on T47D cell lines induced apoptosis which is mediated by the activation of Caspases 8 and 9 and decreased expression of Bcl-2 protein without interference with gen protein 53 (p53). In line with the previous one, tiliroside on MCF7 cell lines induced apoptosis through extrinsic ways due to the underexpression of p53 and Caspase 9 and increase in the expression of Bcl-2 and Caspase 8. Conclusions: The antiproliferative effects of tiliroside are higher on T47D than MCF7 cancers cell lines. Based on molecular analysis, antiproliferative mechanism was by apoptosis through extrinsic pathways.