Purification of hemolysin from Aspergillus fumigatus and study its cytotoxic effect on normal cell line (REF) in vitro

About (20) Pseudomonas rogenosa isolate were experienced for their ability of protease production by calculating the diameter of lysis area after developing on skim milk agar medium (qualitatively ) and the results exhibited that only isolate no (5), was higher isolate for protease making of (26mm) of lysis area. Then, the protein concentration also identified by Bradford method and it was found of 0.16 mg/ ml , then purification was done by using an ion- exchange chromatography with DEAE sephadex G- 100 column and the results showed the presence of 1 peak of enzyme with 50 Kd of molecular weight 2 peaks of other proteins . we tried to investigate the invitro Cytotoxic effect of purified enzyme against two human cancer lines, HeP2 (Human larynx epidermed carcinoma ) , RD ( Rabdo- Sarcoma ) , and one normal cell line Ref ( Rat embryonic fibroblast ) . The cancer and normal cells were treated with different concentrations of protease enzyme ranging from ( 0.05, 0.1, 0.2, 0.4,0.8and 0.16 mg/ml) then incubated for additional 48h at 37C0 and the results showed highest toxicity ( 80.28%) of protease enzyme on RD , moderate cytotoxicity (45.52%) on Hep andslight toxicity ( 37.12% ) on normal cell line (Ref) in a concentration (0.8mg/ml).

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Design, Synthesis and In Vitro Anti-Cancer Evaluation of Novel Derivatives of 2-(2-Methyl-1,5-diaryl-1H-pyrrol-3-yl)-2-oxo-N-(pyridin-3- yl)acetamide

Medicinal Chemistry ◽

10.2174/1573406415666190425153717 ◽

2020 ◽

Vol 16 (3) ◽

pp. 340-349

Author(s):

Ebrahim S. Moghadam ◽

Farhad Saravani ◽

Ernest Hamel ◽

Zahra Shahsavari ◽

Mohsen Alipour ◽

...

Keyword(s):

Cell Cycle ◽

Peripheral Neuropathy ◽

Cell Line ◽

Normal Cell ◽

Caspase 3 ◽

Annexin V ◽

Normal Cell Line ◽

Anti Cancer ◽

Mcf 7

Objective: Several anti-tubulin agents were introduced for the cancer treatment so far. Despite successes in the treatment of cancer, these agents cause toxic side effects, including peripheral neuropathy. Comparing anti-tubulin agents, indibulin seemed to cause minimal peripheral neuropathy, but its poor aqueous solubility and other potential clinical problems have led to its remaining in a preclinical stage. Methods: Herein, indibulin analogues were synthesized and evaluated for their in vitro anti-cancer activity using MTT assay (on the MCF-7, T47-D, MDA-MB231 and NIH-3T3 cell lines), annexin V/PI staining assay, cell cycle analysis, anti-tubulin assay and caspase 3/7 activation assay. Results: One of the compounds, 4a, showed good anti-proliferative activity against MCF-7 cells (IC50: 7.5 μM) and low toxicity on a normal cell line (IC50 > 100 μM). All of the tested compounds showed lower cytotoxicity on normal cell line in comparison to reference compound, indibulin. In the annexin V/PI staining assay, induction of apoptosis in the MCF-7 cell line was observed. Cell cycle analysis illustrated an increasing proportion of cells in the sub-G-1 phase, consistent with an increasing proportion of apoptotic cells. No increase in G2/M cells was observed, consistent with the absence of anti-tubulin activity. A caspase 3/7 assay protocol showed that apoptosis induction by more potent compounds was due to activation of caspase 3. Conclusion: Newly synthesized compounds exerted acceptable anticancer activity and further investigation of current scaffold would be beneficial.

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In vitro anticancer activity of methanolic extract of Granulocystopsis sp., a microalgae from an oligotrophic oasis in the Chihuahuan desert

PeerJ ◽

10.7717/peerj.8686 ◽

2020 ◽

Vol 8 ◽

pp. e8686

Author(s):

Faviola Tavares-Carreón ◽

Susana De la Torre-Zavala ◽

Hector Fernando Arocha-Garza ◽

Valeria Souza ◽

Luis J. Galán-Wong ◽

...

Keyword(s):

Anticancer Activity ◽

Cell Line ◽

Cell Lines ◽

Normal Cell ◽

Chihuahuan Desert ◽

Methanolic Extract ◽

Membrane Integrity ◽

Skin Melanoma ◽

Normal Cell Line

With the purpose of discovering new anticancer molecules that might have fewer side effects or reduce resistance to current antitumor drugs, a bioprospecting study of the microalgae of the Cuatro Cienegas Basin (CCB), an oasis in the Chihuahuan desert in Mexico was conducted. A microalgae was identified as Granulocystopsis sp. through sequencing the rbcL gene and reconstruction of a phylogenetic tree, and its anticancer activities were assessed using various in vitro assays and different cell lines of human cancers, including lung, skin melanoma, colorectal, breast and prostatic cancers, as well as a normal cell line. The values of IC50 of the microalgae methanolic extract using the MTT assay were lower than 20 μg/ml, except that in the lung cancer line and the normal cell line. In vitro, the microalgae extract caused the loss of membrane integrity, monitored by the trypan blue exclusion test and exhibited marked inhibition of adhesion and cell proliferation in cancer cell lines, through the evaluation of the clonogenic assay. Also, typical nuclear changes of apoptotic processes were observed under the microscope, using the dual acridine orange/ethidium bromide fluorescent staining. Finally, the microalgae extract increased the activity of caspases 3 and 7 in skin melanoma, colon, breast and prostate cancer cells, in the same way as the apoptotic inductor and powerful antitumoral drug, doxorubicin. This study shows the anticancer activity from Granulocystopsis sp., a microalgae isolated from the CCB.

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Synthesis, Characterization and Anti-hepatoma Activity of New Hederagenin Derivatives

Mini-Reviews in Medicinal Chemistry ◽

10.2174/1389557519666191010091612 ◽

2020 ◽

Vol 20 (3) ◽

pp. 252-257

Author(s):

Xiao Liu ◽

Lu Sun ◽

Qing-Hua Liu ◽

Bao-Quan Chen ◽

Yu-Ming Liu

Keyword(s):

Hepatocellular Carcinoma ◽

Cell Line ◽

Hepg2 Cell ◽

Normal Cell ◽

Biological Significance ◽

Cancer Cell Line ◽

Human Hepatocellular Carcinoma ◽

Hepg2 Cell Line ◽

Normal Cell Line

Background: Based on the biological significance of hederagenin-type saponins found in our previous investigation, a series of new hederagenin derivatives were designed and synthesized. Methods : Their in vitro antiproliferative activities were evaluated against the HepG2 liver cancer cell line and normal cell line L929 by MTT assay. Results: The preliminary bioassay results demonstrated that all the tested compounds 1-7 showed potent anti-hepatoma activities, and some compounds exhibited better effects than 5-fluorouracil against human hepatocellular carcinoma HepG2 cell line. Furthermore, compound 5 showed a significant antihepatoma activity against HepG2 cells with an IC50 value of 1.88 µM. Besides, all of the tested compounds showed a low cytotoxic effect against the normal cell line L929. Conclusion: All the compounds 1-7 displayed superior selectivity against human hepatocellular carcinoma HepG2 cell line, and the results suggest that the structural modifications of C ring on the hederagenin backbone are vital for modulating anti-hepatoma activities.

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Cytotoxicity Assay of Agaricus Bisporus Extract , Oxaliplatin and Combination of both on Melanoma-B16 and Vero-101 Cell line:An in Vitro study

10.32792/utq/utjsci/vol7/2/3 ◽

2020 ◽

pp. 10-14

Keyword(s):

Cell Line ◽

Crude Extract ◽

Normal Cell ◽

In Vitro Study ◽

Cytotoxicity Assay ◽

Inhibition Rate ◽

Normal Cells ◽

Normal Cell Line ◽

Melanoma B16

The present study included extracted Agaricus bispours and estimated proteins and carbohydrates amount in extract then assessment activity of methanolic crude extract of medicinal mushroom A. bisporus as in vitro anticancer by using melanoma-B16 cell line ,and compared with chemotherapy Oxaliplatin and approved reduction toxic effect of chemotherapy on normal cells when exposed to combination between chemotherapy and mushroom extract on normal vero-101 cell line. Proteins and carbohydrate estimation results referred to there are about 219.33 and 412.98 µg/ml respectively. The results showed that inhibition rate of melanoma-B16 cell line was about 82.15% according to cytotoxicity assay test by crystal violate when treated these cells by methanol crude extract of A. bisporus at 1000 mg/ml .But the cytotoxicity assay of using chemotherapy oxaliplatin on melanoma-B16 at 100 mg/ml results appeared that the inhibition rate was 64.52%. The cytotoxicity assay of combination between methanolic crude extract of A.bisporus 1000 mg/ml and oxaliplatin 100 mg/ml results the inhibition rate was 70.04% on melanoma B-16 cell line.Crystal violate cytotoxicity assay of treatments on normal vero-101 cell line results were revealed that non significantly inhibition of A.bisporus extract on normal cells only 5.37%.While chemotherapy oxaliplatin have inhibition rate on vero-101 normal cell line about 65% ,While the combination treatment cytotoxicity assay on vero-101 normal cell line showed that inhibition rate was reduced to 20.35% comparing with cancer cell which was about 65.3%

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In-vitro Testing of Antioxidant, Anti-Parasite Activities, Cytotoxicity, and Chemical Evaluation of Abutilon Pannosum and Cassia Occidentalis Ethanolic Extracts

Annual Research & Review in Biology ◽

10.9734/arrb/2021/v36i930431 ◽

2021 ◽

pp. 112-121

Author(s):

Ahmed A. Elshikh ◽

Mawahib E. M. ElNour ◽

Hatil H. Elkamali ◽

Ahmed S. Kabbashi

Keyword(s):

Cell Line ◽

Brine Shrimp ◽

Normal Cell ◽

Biological Activities ◽

Ethanolic Extract ◽

Toxicity Tests ◽

Cassia Occidentalis ◽

Normal Cell Line ◽

Ethanolic Extracts

Aims: The aim of this study to detect Anti-giardia, antioxidant activities, cytotoxicity and evaluated the chemical constituent of ethanolic extracts of Abutilon pannosum and Cassia occidentalis. Study Design: Various standard methods were used to detect of bioactivity for ethanolic extracts of plants used in this study. Place and Duration of Study: This study was conducted in the laboratories of microbiology and parasitology and chemistry, the International University of Africa, Khartoum, Sudan, during May 2019. Methodology: The ethanolic extract of Abutilon pannosum and Cassia occidentalis was used as an anti-giardia and anti-oxidant in-vitro, and toxicity tests were performed using brine shrimp and MTT assay. Also, the compounds of the plants used were detected by the GCMS apparatus. Results: The ethanolic extracts of Abutilon pannosum showed high Anti-giardia activity (79%) in concentration (500 ppm) after 72 hours, whereas the activity of Cassia occidentalis extract showed (61%). The highest antioxidant activity of ethanolic extract of Cassia occidentalis was (68.7%), while it was weak in Abutilon pannosum ethanolic extract (45%) by using 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. The results of cytotoxicity revealed that the ethanolic extracts are highly toxic to brine shrimp, but are not toxic to normal cell line (MTT). Chromatographic analysis using gas chromatography-mass spectroscopy (GCMS) showed good separation of compounds. GCMS detected 22 and14 important compounds in Abutilon pannosum and Cassia occidentalis extracts respectively. The common compound in both plant extracts is n-Hexadecanoic acid. This acid was reported as an antioxidant. Conclusion: This study revealed that the biological activities of Abutilon pannosum extracts showed high activities of Anti-giardia and antioxidants. Non-cytotoxic in the normal cell line was shown. Cassia occidentalis showed high activity of Anti-giardia and weak activity antioxidant.

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In vitro biocompatibility evaluation of biscoumarin based random copolyesters

New Journal of Chemistry ◽

10.1039/c5nj00322a ◽

2015 ◽

Vol 39 (6) ◽

pp. 4948-4956 ◽

Cited By ~ 5

Author(s):

Kandaswamy Narendran ◽

Raveendiran Nanthini

Keyword(s):

Cell Line ◽

Normal Cell ◽

Vero Cells ◽

Normal Cell Line ◽

In Vitro Biocompatibility

Copolyester CP5 exhibits cytocompatible properties toward a normal cell line (Vero cells) and requires 13-fold higher concentration in comparison with Hep-2 cells.

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Evaluation for the Effect of Heat Stable Enterotoxin (a) Produced by Enterotoxigenic Escherichia coli on Different Cancer Cells In Vitro

Baghdad Science Journal ◽

10.21123/bsj.7.1.401-410 ◽

2010 ◽

Vol 7 (1) ◽

pp. 401-410

Author(s):

Baghdad Science Journal

Keyword(s):

Cancer Patients ◽

Cancer Cell ◽

Cell Cultures ◽

Normal Cell ◽

Cytotoxic Effect ◽

Enterotoxigenic Escherichia Coli ◽

Primary Cancer ◽

Normal Cell Line ◽

Heat Stable

This study was conducted for evaluating the cytotoxic effect of heat stable enterotoxin a (STa) produced by enterotoxigenic Escherichia coli on the proliferation of primary cancer cell cultures, obtained from tumor samples that were collected from (13) cancer patients and as follows: (five colon cancer patients, two bladder cancer patients, two breast cancer patients, two stomach cancer patients and two lung cancer patients), and on normal cell line (rat embryonic fibroblast / REF) (in vitro) with the use of different concentrations starting from (1) mg/ml and ending with (0.0002) mg/ml by making two fold serial dilutions by using the 96- well microtiter plate, and in comparison with negative (PBS) and positive (MMC, at concentration of 10 µg/ml) controls . Results showed that, after (24) hours of exposure to STa, the growth of all primary cancer cell cultures obtained from colon cancer patients was inhibited by STa treatment and this inhibition was concentration dependent. Also it was shown that the cytotoxic effect of the high concentration of STa was close to that seen after MMC treatment. While no differences were seen in the growth of all primary cancer cell cultures that were obtained from the other cancer patients, which mean that STa treatment neither inhibit nor enhanced their growth. At the same time STa did not show or has any cytotoxic effect on the normal cell line (REF).

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The effect of crude methanolic extract of ginger (Zingibe officinale) root in different cell lines in vitro

Journal of Biotechnology Research Center ◽

10.24126/jobrc.2009.3.1.56 ◽

2009 ◽

Vol 3 (1) ◽

pp. 122-129

Author(s):

Shamaa Ismael Kadhum ◽

Safaa Al-deen Ahmed Alqysi

Keyword(s):

Cell Line ◽

Cell Lines ◽

Cancer Cell ◽

Plant Extract ◽

Normal Cell ◽

Methanolic Extract ◽

Cancer Cell Lines ◽

The Other ◽

Normal Cell Line

This study involved the affectivity of crude methanolic extract of ginger root on different cells line in vitro, four cancer cell lines were tested Hela, L20B,Hep2, AMN3 compared with normal cell line (REF)and transformed cell line (Vero). Four extract concentrations were prepared (125,250,500,1000) µg/ml respectively, the results showed a significant inhibitory effect on the growth of different cell lines under study, also regression showed a significant negative relationship between plant extract and cell lines,1000 µg/ml concentration showed significant effect on cell lines growth (HELA,Hep2,L20B and Vero) on the other hand AMN3 was not affected by the plant extract, there was a direct relationship between concentrations and the rate of inhibition of the cell lines, on the other hand the normal cell line were more effected than cancer cell lines under study.

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Novel 1,2,3-Triazole-functionalized pyrido[3',2':4,5]furo[3,2-d]pyrimidin- 4(3H)-one Derivatives: Synthesis, Anticancer Activity, CoMFA and CoMSIA Studies

Letters in Organic Chemistry ◽

10.2174/1570178617666200319124017 ◽

2020 ◽

Vol 17 (12) ◽

pp. 969-978

Author(s):

Balakishan Vadla ◽

Sailu Betala

Keyword(s):

Anticancer Activity ◽

Cell Line ◽

Normal Cell ◽

Rational Design ◽

Human Cancer ◽

Strong Basis ◽

Hek 293 ◽

Normal Cell Line ◽

Human Cancer Cell Lines ◽

Mcf 7

A series of novel triazole functionalized pyrido [3',2':4,5] furo[3,2-d] pyrimidin-4 (3H)-one derivatives 7a-p were prepared from ethyl furo[2,3-b]pyridine-2-carboxylate 3 on reaction with ammonia to afford furo[2,3-b]pyridine-2-carboxamide 4. This compound, on reaction with triethyl orthoformate TEOF, gave compound 5. Compound 5 on propargylation, followed by a reaction with substituted aryl azides under Sharpless reaction conditions, furnished triazole tagged pyrido [3',2':4,5]furo[3,2-d] pyrimidin-4(3H)-one derivatives. All the products 7a-p were screened against four human cancer cell lines, such as HeLa - Cervical cancer (CCL-2), COLO 205- Colon cancer (CCL-222), HepG2- Liver cancer (HB-8065), and MCF7 - Breast cancer (HTB-22) and one normal cell line (HEK 293). Compounds 7b, 7n, 7o and 7p, which showed promising anticancer activity, were identified and found to be non-toxic to normal cell line. Studies for HeLa, COLO205, HepG2, and MCF-7 using CoMFA and CoMSIA were carried out . Models from 3D-QSAR provided a strong basis for future rational design of more active and selective HeLa, COLO205, HepG2, and MCF-7 cell line inhibitors.

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