scholarly journals CYTOTOXIC ACTIVITY AND APOPTOSIS INDUCTION OF T47D CELL LINES BY Turbinaria decurrens EXTRACT

Author(s):  
Muhammad Nursid ◽  
Nurrahmi Dewi Fajarningsih ◽  
Ekowati Chasanah

Marine algae is known to contain a wide variety of biomedical compounds having pharmaceutical applications. The aim of this research was to evaluate cytotoxic activity and apoptosis induction of Turbinaria decurrens extract on T47D cell lines.  Cytotoxic activity test was conducted by using MTT assay whereas detection of apoptosis was evaluated by DNA fragmentations and flow cytometry analysis. The MTT test showed that crude extract had medium cytotoxic activity to T47D, HepG2, and C28 cell lines with IC50 value of 172, againts 360 and 330 µg ml-1, respectively. After solvent partition of crude extract, the cytotoxic activity of n-hexane and ethyl acetate fractions T47D cell increased, the cytotoxic activity of n. hexane and ethyl acetate fractions T47D cell increased with  IC50  value of with IC50  43.1 and 51.9 µg ml-1, respectively, whereas IC50 value of methanol fraction was 383.0 µg ml-1. Analysis of DNA fragmentation of T47D cell showed that  both n-hexane and ethyl acetate fractions could not fragment DNA as a features of apoptosis. However, flow cytometry analysis by using annexin-V and propidium iodide staining revealed that n-hexane and ethyl acetate fractions could induce apoptosis in T47D cell. This research indicated that Turbinaria decurrens had potency to induce apoptosis in T47D cells.

Author(s):  
Poppy Anjelisa Zaitun Hasibuan ◽  
Panal Sitorus ◽  
Denny Satria ◽  
Rizka Damela Sibuea

Research into plants with anticancer effects is actively encouraged in orderto discover new drugs with lessertoxicity but more potent effects. The aims of study are to evaluate the antioxidant properties and to investigate the cytotoxic activity of Plectranthus amboinicus (Lour.) Spreng. leaves ethyl acetate fractions on HeLa,T47D and MCF7 cell lines. The extract was prepared by graded maceration using n-hexane and ethyl acetate. The ethyl acetate extract was fractionated in vacuum liquid chromatography with n-hexane: ethyl acetate; and ethyl acetate: methanol as mobile phase. Then, the fractions were analyzed with thin layer chromatography (TLC). The free radical scavenging activity was measured by DPPH method, the total flavonoid content was calculated by quercetin equivalent and the absorbance is measured by using UV-Visible spectrophotometry. The cytotoxic activity were determined using MTT assay. The fractions contained 5 sub fractions with same TLC profile. The fractions showed antioxidant activity by DPPH method with different IC50 values, namely: 130 µg/mL(I), 127 µg/mL(II), 137 µg/mL(III), 129 µg/mL(IV), and 124 µg/ mL(V), respectively. The measurement of total flavonoid content showed 118 mg QE/g (I), 50 mg QE/g (II), 207 mg QE/g (III), 56 mg QE/g (IV), and 55 mg QE/g (V). The IC50 of each sub fractions on HeLa cell were 77 µg/mL, 46 µg/mL, 93 µg/mL, 71 µg/mL and 476 µg/mL; for T47D cell were 1621 µg/mL, 111 µg/mL, 128 µg/mL, 150 µg/mL and 209 µg/mL; and for MCF7 were 259 µg/mL, 343 µg/mL, 575 µg/mL, 408 µg/mL and 250 µg/mL. Based on the results, the fractions derived from ethyl acetate extract of Plectranthus amboinicus (Lour.) Spreng. leaves exhibit antioxidant. The Fraction II from ethyl acetate extract of Plectranthus amboinicus (Lour.) Spreng. was the most cytotoxic on HeLa, T47D and MCF7 cell lines. It is potential to undergo further isolation of its cytotoxic compounds.Keywords : antioxidant, cytotoxic, Plectranthus amboinicul (Lour.) Spreng., ethyl acetate fractions


Author(s):  
Muhammad Da'i ◽  
Khairunnisa Azani Meilinasary ◽  
Andi Suhendi ◽  
Sari Haryanti

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


Author(s):  
Andi Suhendi ◽  
Erindyah Retno Wikantyasning ◽  
Gunawan Setyadi ◽  
Arifah Sri Wahyuni ◽  
Muhammad Da'i

Due 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.Key words: Alpinia galanga, HeLa, T47D, MCF7 and Acetoxy chavicol acetate


2014 ◽  
Vol 2 (2) ◽  
pp. 53-58 ◽  
Author(s):  
Iin Tusanti ◽  
Andrew Johan ◽  
RA Kisdjamiatun

Background: Several studies focused on phytochemical as agents of cancer prevention and co-chemotherapy. One of Indonesian plant which has edible fruit but it hasn’t been completely explored is Medinilla speciosa (Reinw.ex Bl.). Objective : The aim of this study is to examine the cytotoxic activity (IC50 value) of Medinilla speciosa (Reinw.ex Bl.) fruit ethanolic extract. Methods : Medinilla speciosa (Reinw.ex Bl.) fruit ethanolic extract was used in this study. The cytotoxic activity was investigated in vitro on human breast cancer T47D cell-line. The cells viability were assessed using MTT colorimetric assay. Breast cancer T47D cell lines was treated with fruit ethanolic extract (10, 25, 50, 100, 250, 500 and 1000 µg/ml) for 24 hour of incubation. This study also identified phytochemical compound of the fruit with thin layer chromatography (TLC). Results: The result showed that ethanolic extract of Medinilla speciosa (Reinw.ex Bl.) has moderate cytotoxicity on breast cancer T47D cell line with IC50 value of 614.50 µg/ml and yield the decrease of cell viability at higher concentration. Medinilla speciosa fruit can not be used as anticancer agent but chemoprevention agent. Phytochemical test showed that the fruit extract contain flavonoid and saponin compound. Conclusion: Ethanolic extract of Medinilla speciosa fruit exhibited moderate cytotocicity on breast cancer T47D cell lines with IC50 value was 614,50 µg/ml thus it can be used as chemopreventioan agent.


Author(s):  
Zeinab Abedian ◽  
Niloofar Jenabian ◽  
Ali Akbar Moghadamnia ◽  
Ebrahim Zabihi ◽  
Roghayeh Pourbagher ◽  
...  

Objective/ Background: Cancer is still the most common cause of morbidity in world and new powerful anticancer agents without severe side effects from natural sources is important. Methods: The evaluation of cytotoxicity and apoptosis induction was carried out in MCF-7,HeLa and Saos-2 as cancerous cell lines with different histological origin and human fibroblast served as control normal cell. The cells were treated with different concentrations of chitosan and the cytotoxicity was determined using MTT assay after 24, 48 and 72 h .The mode of death was evaluated by flow cytometry . Results: While both types of chitosan showed significant concentration-dependently cytotoxic effects against the three cancerous cell lines, fibroblast cells showed somehow more compatibility with chitosan. On the other hand, there were no significant differences between LMWC and HMWC cytotoxicity in all cell lines. The flow cytometry results showed the apoptosis pattern of death more in Saos-2 and HeLa while necrosis was more observable with MCF7. Also higher viability with both types of chitosan was seen in fibroblast as normal cells Conclusion: Chitosan shows anticancerous effect against 3 cancerous cell lines, while it is compatible with normal diploid fibroblast cells. Furthermore, it seems that the molecular weight of chitosan does not affect its anticancerous property.


2021 ◽  
Vol 11 (11) ◽  
pp. 5300
Author(s):  
Jozef Hudec ◽  
Jan Mojzis ◽  
Marta Habanova ◽  
Jorge A. Saraiva ◽  
Pavel Hradil ◽  
...  

Sarcopoterium spinosum (L.) is a medicinal plant traditionally used for the treatment of various diseases including cancer in the Near- and Middle East. The fractions and constituents of the ethanol extract of S. spinosum were screened for in vitro cytotoxic activities on Jurkat (acute T-lymphoblastic leukemia), HeLa (cervical adenocarcinoma), MCF-7 (mammary gland adenocarcinoma), Caco-2 (human colorectal adenocarcinoma), and MDA-MB-231 (mammary gland adenocarcinoma) cell lines using the MTT (3-(dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. The ethanol extract was subsequently re-extracted with ethyl acetate and in its sub-fraction obtained by column chromatography three compounds (stachydrine, benzalkonium chloride and rutine) were the first time identified by nuclear magnetic resonance (NMR) analyses. The most active subfraction showed cytotoxic activity against HeLa, MCF-7, and Caco-2 cell lines. The three compounds mentioned, as standards of high-performance liquid chromatography (HPLC) quality, were studied individually and in combination. Cytotoxic activity observed might be due to the presence of benzalkonium chloride and rutin. Benzalkonium chloride showed the strongest growth suppression effect against HeLa cells (IC50 8.10−7 M) and MCF-7 cells (IC50 5.10−6 M). The mixture of stachydrine and benzalkonium chloride allowed a synergistic cytotoxic effect against all tested cancer and normal cells to be obtained. Anti-cancer activity of the plant extract of S. spinosum remains under-investigated, so this research describes how the three major compounds identified in the ethyl acetate extract can exert a significant dose dependent in vitro cytotoxicity.


2014 ◽  
Vol 5 (7) ◽  
pp. 557-559 ◽  
Author(s):  
Elenora Kristanty Ruth ◽  
Suriawati Junie ◽  
Sulistiyo Joko

2000 ◽  
Vol 55 (1-2) ◽  
pp. 40-43 ◽  
Author(s):  
Miguel López-Lázaro ◽  
Carmen Martín-Cordero ◽  
Felipe Cortés ◽  
Joaquín Piñero ◽  
María Jesús Ayuso

Abstract Seven flavonoids isolated from chloroform , ethyl acetate and butanol extracts, obtained from the aerial parts of Retama sphaerocarpa, have been assessed for cytotoxic activity against three human cancer cell lines: TK-10 (renal adenocarcinom a), MCF-7 (breast adenocarcinom a) and UACC-62 (m elanom a), using the SRB assay. All of them , extracts and flavonoids, were actives in, at least, one of the three cell lines at the recom m ended N ational C ancer Institute doses. They produce a d ose-dependent inhibition of cell grow th at concentrations in the 10-6-10-4 ᴍ and 25 -250 μg/ml range for the flavonoids and extracts respectively, being the flavonol rhamnazin the most cytotoxic.


2012 ◽  
Vol 2012 ◽  
pp. 1-9 ◽  
Author(s):  
Faizah Bt. Harun ◽  
Syed Mohsin Syed Sahil Jamalullail ◽  
Khoo Boon Yin ◽  
Zulkhairi Othman ◽  
Anita Tilwari ◽  
...  

Eupatorium odoratum (EO)contains many biologically active compounds, the anticancer effects of which are not well documented. This study evaluates the cytotoxic effects and mechanism of action ofEOextracts on MCF-7 and Vero cell lines. Evaluation of the cytotoxic activity using MTT assay, morphological alterations, and apoptosis were carried out. Autophagy was evaluated by LC3-A protein expression. Cytotoxic activity, membrane blebbing and ballooning at 24 hours, replacement by mass vacuolation, and double membrane vesicles mimicking autophagy and cell death were observed in the cancer cells. No apoptosis was observed by DNA fragmentation assay. Overexpression of LC3-A protein indicated autophagic cell death. Cell cycle analysis showed G0 and G2/M arrest. The Vero cells did not show significant cell death at concentrations <100 μg/mL. These results thus suggest that acetone and ethyl acetate extracts ofEOinduce cell death through induction of autophagy and hold potential for development as potential anticancer drugs.


Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 3023-3023
Author(s):  
Miyuki Ookura ◽  
Tatsuya Fujii ◽  
Shinji Kishi ◽  
Hiroko Shigemi ◽  
Naoko Hosono ◽  
...  

Abstract Multiple myeloma (MM) is a molecularly heterogeneous hematologic malignancy and remains mostly incurable despite the recent improvement of treatment strategies by several novel agents. Therefore, it is important to develop more efficacious drug against MM. YM155, a novel small molecule suppressant of survivin, shows anti-proliferative activities against various human cancer cells. YM155 was identified in a survivin gene promoter assay by high throughput screening of chemical libraries. In the present study, we investigated the cytotoxic mechanism of YM155 against human myeloma cells including bortezomib (BTZ) resistant cells (U266/BTZ). Three myeloma cell lines, U266, KMS-11 and KMS-12, were employed. YM155 inhibited the cell growth of these cell lines with the IC50 value of below 5 nM. YM155 suppressed the expression of mRNA and protein of survivin. We also found that YM155 inhibited the protein expression of Mcl-1, as an essential anti-apoptotic protein for survival of myeloma cells. In addition, we observed that YM155 markedly suppressed the phosphorylation of STAT3, which is known as transcription factor of Mcl-1. When KMS-12 cells were incubated with IL-6, phosphorylation of STAT3 and upregulation of Mcl-1 protein were observed. Treatment of KMS-12 with YM155 inhibited these events and eventually induced apoptosis in KMS-12 cells. Interestingly, inhibitory effect of YM155 on Mcl-1 protein expression was much stronger than that on survivin. RQ-PCR analysis indicated that the level of Mcl-1 mRNA was not affected after YM155 treatment. Immunoblot analysis showed that proteasome inhibitor MG-132 blocked the inhibition of Mcl-1 expression by YM155, suggesting that proteasome-mediated degradation is involved in YM155-induced Mcl-1 downregulation. MM is a low-growth-fraction disease and low proliferation of MM seems to contribute to resistance to various anticancer drugs. To determine whether YM155 shows cytotoxic effect against quiescent (G0/G1) MM cells, U266 were cultured in low-serum medium to enrich the G0/G1 population. Dual-parametric flow cytometric analysis using Hoechest33342 and the RNA specific dye pyronin Y revealed that YM155 potently induced cell death of quiescent (G0/G1) MM cells. In quiescent MM cells, inhibitory effect of YM155 on Mcl-1 protein expression was much stronger than that on survivin. We also examined whether similar effect of YM155 could be observed in primary MM cells. The majority of primary MM cells from patients was found to be in quiescent phase by cell-cycle analysis. YM155 showed similar cytotoxic activity against primary MM cells. In contrast, Ara-C, the S-phase specific anticancer drug, never killed quiescent primary MM cells. We established BTZ-resistant MM cell line (U266/BTZ). The IC50 value was 45-fold higher than its parental cell line. DNA sequencing data indicated that U266/BTZ cells possess a point mutation, G322A, in the gene encoding the proteasome beta-5 subunit. YM155 almost equally exhibited cytotoxic activity against U266/BTZ compared with parental cells. U266/BTZ displayed significantly lowered amounts of bcl-2, survivin and aurora-B kinase proteins. Interestingly, U266/BTZ overexpressed the Mcl-1 protein. Treatment with YM155 rapidly suppressed Mcl-1 protein expression and induced apoptosis. These data suggest that overexpression of Mcl-1 may contribute to bortezomib resistance and downregulation of Mcl-1 by YM155 could overcome it. In conclusion, our data indicate that YM155 may exert robust cytotoxic activity against quiescent (G0/G1) MM and bortezomib resistant cells via inhibition of Mcl-1 and survivin. Disclosures No relevant conflicts of interest to declare.


Sign in / Sign up

Export Citation Format

Share Document