scholarly journals Evaluation of cytotoxic potential of L-asparaginase from Scopulariopsis brevicaulis on cell lines in vitro

Biomedicine ◽  
2020 ◽  
Vol 39 (2) ◽  
pp. 254-257
Author(s):  
D’Souza Renita Maria ◽  
Abraham Asha

Introduction and Aim: This study reports the cytotoxic potential of L-Asparaginase isolated from the fungus Scopulariopsis brevicaulis. Materials and Methods: Extracellular L- Asparaginase was isolated from Scopulariopsis brevicaulis and purified by ammonium sulfate precipitation, followed by dialysis, ion exchange and gel filtration chromatography. Varying concentrations (31.25, 62.5, 125, 250, 500 µg/ml) of purified L-Asparaginase was tested on MCF7, HeLa, HepG2 and 3T3L1cell lines by MTT assay. Curcumin was maintained as a positive control. Results: Minimum inhibition of 23.57% was observed at an enzyme concentration of 31.25 µg/ml and maximum inhibition (66.41%) was observed at 500 µg/ml against MCF7 cell line. Minimum inhibition of 2.87% was observed at an enzyme concentration 31.25 µg/ml and maximum inhibition (58.49%) was observed at 500 µg/ml against HeLa cell line. Minimum inhibition of 4.58% was shown at an enzyme concentration of 31.25 µg/ml and maximum inhibition (46.14 %) was observed at 500 µg/ml against HepG2 cell line. Minimum inhibition of 1.4% was shown by enzyme concentration 31.25 µg/ml and maximum inhibition (50.9%) was observed at 500 µg/ml against 3T3L1 cell line. Conclusion: We report for the first time the cytotoxic potential of L-Asparaginase from Scopulariopsis brevicaulis.  

2019 ◽  
Vol 2 (2) ◽  
pp. 49 ◽  
Author(s):  
Renita Maria D'Souza ◽  
Asha Abraham

This study reports the cytotoxic potential of L-Asparaginase isolated from Bacillus sp. Bacillus sp. was isolated from local soil/water samples and identified by rapid plate assay and further confirmed by phenotypic characterization. Extracellular L- Asparaginase was isolated from broth culture of Bacillus sp.  and purified by ammonium sulfate precipitation, followed by dialysis and ion exchange and gel filtration chromatography techniques. The purified enzyme was used to study the in vitro cytotoxic potential. Varying concentrations (31.25, 62.5, 125, 250, 500 µg/ml) of purified L-Asparaginase was tested on MCF7, HeLa, HepG2 and 3T3L1cell lines by MTT assay. Curcumin was maintained as a positive control. The results revealed that the enzyme showed a significant cytotoxic activity and a dose dependent effect. Minimum inhibition of (19.44%) was observed at an enzyme concentration of 31.25 µg/ml and maximum inhibition (71.14%) was observed at 500ug/ml against MCF7 cell line. Minimum inhibition of (10.04%) was observed at an enzyme concentration 31.25 µg/ml and maximum inhibition (68.92%) was observed at 500 µg/ml against HeLa cell line. Minimum inhibition of (7.45%) was shown at an enzyme concentration of 31.25 µg/ml and maximum inhibition (68.28 %) was observed at 500 µg/ml against HepG2 cell line. Minimum inhibition of (4.4%) was shown by enzyme concentration 31.25 µg/ml and maximum inhibition (47.4%) was observed at 500 µg/ml against 3T3L1 cell line. Curcumin at 5µM concentration showed an inhibition of 48.13% against MCF-7 cells, 54.42% against HeLa cells, 64.94% against HepG2 cells and 44.5% against 3T3L1 cell lines.


Molecules ◽  
2020 ◽  
Vol 25 (4) ◽  
pp. 888 ◽  
Author(s):  
Szabolcs Mayer ◽  
Péter Keglevich ◽  
Péter Ábrányi-Balogh ◽  
Áron Szigetvári ◽  
Miklós Dékány ◽  
...  

Chrysin is a naturally occurring flavonoid with mild anticancer activity. In this paper we report the synthesis of new chrysin derivatives alkylated with N-phenylchloroacetamides in position 7. A novel method was developed for the preparation of 7-aminochrysin derivatives via the Smiles rearrangement, resulting in diphenylamine-type compounds. In silico studies of the Smiles rearrangement were performed. We also present the in vitro antiproliferative activity of the synthesized compounds against 60 human tumor cell lines (NCI60). The most potent derivative exhibited nanomolar antitumor activity on the MCF7 cell line of breast cancer (GI50 = 30 nM) and on the HCT-15 cell line of colon cancer (GI50 = 60 nM).


2019 ◽  
Vol 17 (1) ◽  
pp. 943-954
Author(s):  
Abeer N. Al-romaizan ◽  
Thoraya S. Jaber ◽  
Nesreen S. Ahmed

AbstractA series of new 2-phenyl-7-methyl-1,8-naphthyridine derivatives with variable substituents at C3 were synthesized for an in vitro evaluation of their anticancer activity against human breast cancer cell line (MCF7). On one hand, compounds 3f, 6f, 8c, and 10b showed IC50 values (6.53, 7.88, 7.89, 7.79 μM, respectively) compared to that of the mentioned drug staurosparine (IC50 = 4.51 μM). On the other hand, derivatives 10c, 8d, 4d, 10f and 8b displayed better activity than staurosporin with IC50 values (1.47, 1.62, 1.68, 2.30, 3.19 μM, respectively).


2019 ◽  
Vol 43 (3-4) ◽  
pp. 130-134
Author(s):  
Shao-Rui Chen ◽  
Hao Wu ◽  
Hai-Yan Zhao ◽  
Yu-Mei Zhang ◽  
Peng-Qi Li ◽  
...  

A new series of 4-azasteroidal-17-hydrazone derivatives have been synthesized from androstenedione. Their structures were characterized by analysis and spectroscopic data. The antiproliferative activity of synthesized compounds against three cancer cells (human lung adenocarcinoma, human oesophageal cervical cancer, human gastric adenocarcinoma) and a normal cell line (human gastric mucosal) was investigated. The studies show that the compound bearing a naphthyl group displayed the same antiproliferative activity in vitro against tested cells as cis-platin did (a positive control). Most of the compounds show very weak toxicity towards normal human gastric mucosal cell line.


Parasitology ◽  
2009 ◽  
Vol 136 (8) ◽  
pp. 895-904 ◽  
Author(s):  
S. RATHAUR ◽  
R. RAI ◽  
E. SRIKANTH ◽  
S. SRIVASTAVA

SUMMARYSetaria cervi, a bovine filarial parasite contains significant acid phosphatase (AcP) activity in its various life stages. Two forms of AcP were separated from somatic extract of adult female parasite using cation exchange, gel filtration and concavalin affinity chromatography. One form having a molecular mass of 79 kDa was characterized as dual specific protein tyrosine phosphatase (ScDSP) based on substrate specificity and inhibition studies. With various substrates tested, it showed significant activity in the order of phospho-L-tyrosine>pNPP>ADP>phospho-L-serine. Inhibition by orthovanadate, fluoride, molybdate, and zinc ions further confirms protein tyrosine phosphatase nature of the enzyme. Km and Vmax determined with various substrates were found to be 16·66 mM, 25·0 μM/ml/min with pNPP; 20·0 mM, 40·0 μM/ml/min with phospho-L-tyrosine and 27·0 mM, 25·0 μM/ml/min with phospho-L-serine. KIwith pNPP and sodium orthovanadate (IC5033·0 μM) was calculated to be 50·0 mM. Inhibition with pHMB, silver nitrate, DEPC and EDAC suggested the presence of cysteine, histidine and carboxylate residues at its active site. Cross-reactivity withW. bancrofti-infected sera was demonstrated by Western blotting. ScDSP showed elevated levels of IgE in chronic filarial sera using ELISA. Underin vitroconditions, ScDSP resulted in increased effector function of human eosinophils when stimulated by IgG, which showed a further decrease with increasing enzyme concentration. Results presented here suggest thatS. cerviDSP should be further studied to determine its role in pathogenesis and the persistence of filarial parasite.


2001 ◽  
Vol 382 (2) ◽  
pp. 291-297 ◽  
Author(s):  
Stephan Hinderlich ◽  
Markus Berger ◽  
Oliver T. Keppler ◽  
Michael Pawlita ◽  
Werner Reutter

Abstract The first two steps in mammalian biosynthesis of Nacetylneuraminic acid, an important carbohydrate moiety in biological recognition systems, are performed by the bifunctional enzyme UDPNacetylglucosamine 2-epimerase/Nacetylmannosamine kinase. A subclone of the human B lymphoma cell line BJAB K20, lacking UDPNacetylglucosamine 2- epimerase/Nacetylmannosamine kinase mRNA as well as epimerase activity, displayed hyposialylated, functionally impaired cell surface glycoconjugates. Here we show that this cell line surprisingly still retains Nacetylmannosamine kinase activity. A gel filtration analysis of BJAB K88 control cells, which express UDPNacetylglucosamine 2-epimerase/Nacetylmannosamine kinase, revealed two Nacetylmannosamine kinase activity peaks, one coeluting with UDPNacetylglucosamine 2-epimerase activity and one coeluting with Nacetylglucosamine kinase. For this enzyme previous studies already showed ManNAc kinase activity in vitro. In contrast, the hyposialylated BJAB K20 subclone displayed only the Nacetylmannosamine kinase peak, comigrating with Nacetylglucosamine kinase. The CMPNacetylneuraminic acid content of both K88 and K20 cells and the sialylation of cell surface glycoconjugates of K20 cells could be significantly increased by supple menting the medium with Nacetylmannosamine. This Nacetylmannosamineinduced increase was drastically reduced by cosupplementation with Nacetylglucosamine only in K20 cells. We therefore propose the phosphorylation of Nacetylmannosamine as a hitherto unrecognized role of Nacetylglucosamine kinase in living cells.


2007 ◽  
Vol 85 (1) ◽  
pp. 88-95 ◽  
Author(s):  
Amandeep Kaur ◽  
Sukhdev Singh Kamboj ◽  
Jatinder Singh ◽  
Rajinder Singh ◽  
Melissa Abrahams ◽  
...  

Three monomeric monocot lectins from Zephyranthes carinata, Zephyranthes candida, and Gloriosa superba with carbohydrate specificity towards mannose derivatives and (or) oligomannose have been isolated and purified from their storage tissues. The lectins were purified by anion-exchange chromatography on DEAE–Sephacyl and by gel filtration chromatography on Biogel P-200 followed by high-performance liquid chromatography. The purified lectins, Z. carinata, Z. candida, and G. superba had molecular masses of 12, 11.5, and 12.5 kDa, respectively, as determined by gel filtration and SDS–PAGE, indicating that they are monomers. In a hapten inhibition assay, methyl-α-d-mannopyranoside inhibited agglutination of both Z. candida and Z. carinata; the latter was also inhibited by Man(α1-2)Man and Man(α1-3)Man. Gloriosa superba showed inhibition only with Man(α1-4)Man of all of the sugars and glycoproteins tested. All purified lectins agglutinated red blood cells from rabbit, whereas G. superba was also reactive towards erythrocytes from guinea pig. All of the lectins were nonglycosylated and did not require metal ions for their activity. They were labile above 60 °C and were affected by denaturing agents such as urea, thiourea, and guanidine–HCl. The lectins were virtually nonmitogenic, like other members of Amaryllidaceae and Liliaceae. Of the 3 lectins, G. superba was found to be highly toxic to the BSC-1 cell line (African green monkey kidney epithelial cells), while both of the Zephyranthes species showed significant in vitro inhibition of poxvirus replication in BSC-1 cells without any toxic effects to the cells. In addition, Z. candida also exhibited significant anticancer activity against SNB-78, a CNS human cancer cell line.


PeerJ ◽  
2018 ◽  
Vol 6 ◽  
pp. e4373 ◽  
Author(s):  
Xueqin Wang ◽  
Huahua Yu ◽  
Ronge Xing ◽  
Xiaolin Chen ◽  
Song Liu ◽  
...  

Mackerel (Pneumatophorus japonicus) viscera contain large amount of protein. We used five proteases to hydrolyze the viscera, and the hydrolysate treated by neutrase exhibited the highest nitrogen recovery (NR). Then we optimized the preparation conditions for mackerel viscera hydrolysate (MVH) by response surface methodology and investigated the antioxidant activity of MVH. The optimal conditions were as follows: enzyme concentration of 1,762.87 U/g, pH of 6.76, temperature of 43.75 °C, extraction time of 6.0 h and water/material ratio of 20.37 (v/w), and the maximum NR was 37.84%. Furthermore, the molecular weight distribution of MVH was almost below 3,000 Da determined by TSK G2000 SWXL gel filtration chromatography, and the MVH exhibited good antioxidant activities in various in vitro assays, including DPPH radical, hydroxyl radical and superoxide anion scavenging activities, reducing power and similar effectivelness as butylated hydroxytoluene and Vitamin E to inhibit lipid peroxidation. The results suggested that MVH could be used as a potential source of antioxidant peptide in food industries.


2018 ◽  
Vol 24 (S) ◽  
pp. 853-858
Author(s):  
Shazia Kanwal Malik ◽  
Maqsood Ahmed ◽  
Farah Khan

Background: Medicinal plants are used in anticancer therapies from ages. The very first time we have not only screened anticancer medicinal plants from Pakistan but also identified anticancer plant terpenes. This study will definitely pave the way towards anticancer drug development after animal trials. Materials and Methods | This study was based on ethanolic extraction of fruits/seed of 25 plants. The cytotoxic potential of extracts and plant terpenes was evaluated against mouse fibroblasts (NIH 3T3) cell line by using MTT (3- [4, 5-dimethylthiazole-2-yl]-2, 5-diphenyl-tetrazolium bromide) assay and compared with cyclohexamide. Results | Ethanolic extracts were nontoxic against NIH3T3 cell line. On the basis of the % inhibition plants were graded as B.lycium, 48% > T.bellerica, 47% > C.verum, 45% > Z.tenuior, 39% > H. adenophyllum, 38% > C. carandas, 31% > C.behen, 29% > P. juliflora, 28% & P.granatum, 28% W.somnifera, 27% > E.cheiri, 26% > S. potatorum, 24% & M.incana, 24% > D. peregrina, 21% & B.lanzan, 21% > L. maldivica, 20% > F. lyrata, 18% > F.arabica, 16% & R. indica, 16% > C.absus, 13% & C.paniculatus, 13%. > C. diurnum, 12% > J.mimosifolia, 11% > D.malabarica, 8% > C. speciosa, 3% as compared to the reference drug Cyclohexamide, 70%. Whereas the maximum % inhibition was exhibited by n-Hexadecanoic acid (88%), followed by Hexadecanoic acid, methyl ester (16%) and Methyl decanoate displayed 11%. C50 of n-Hexadecanoic acid and Cyclohexamide was 3.1±0.2 and 0.8±0.2 respectively. Conclusion| Plant extracts and terpenes was found to be inactive against 3T3 cell line.


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