Dear Aliyu,

You might be assisted with the following publication for the determination of the cytotoxicity effect of the endophytes you isolated from roots of plants:

Important Paper: (see attached file)

Antonie Van Leeuwenhoek. 2015; 108(2): 267–289.

Published online 2015 Jun 21. doi:  10.1007/s10482-015-0502-7

PMCID: PMC4491368

Endophytic actinobacteria of medicinal plants: diversity and bioactivity

Patrycja Golinska, Magdalena Wypij, Gauravi Agarkar, Dnyaneshwar Rathod, Hanna Dahm, and Mahendra Rai

Author information ► Article notes ► Copyright and License information ►

Go to:

Abstract

Endophytes are the microorganisms that exist inside the plant tissues without having any negative impact on the host plant. Medicinal plants constitute the huge diversity of endophytic actinobacteria of economical importance. These microbes have huge potential to synthesis of numerous novel compounds that can be exploited in pharmaceutical, agricultural and other industries. It is of prime importance to focus the present research on practical utilization of this microbial group in order to find out the solutions to the problems related to health, environment and agriculture. An extensive characterization of diverse population of endophytic actinobacteria associated with medicinal plants can provide a greater insight into the plant-endophyte interactions and evolution of mutualism. In the present review, we have discussed the diversity of endophytic actinobacteria of from medicinal plants their multiple bioactivities.

Keywords: Actinobacteria, Antimicrobial activity, Bioactive compounds, Endophytes, Medicinal plants

1- Pharm Biol. 2012 Nov;50(11):1416-22. doi: 10.3109/13880209.2012.682118. Epub 2012 Sep 11.

Cytotoxicity effects of various Juglans regia (walnut) leaf extracts in human cancer cell lines.

Salimi M1, Majd A, Sepahdar Z, Azadmanesh K, Irian S, Ardestaniyan MH, Hedayati MH, Rastkari N.

Author information

Abstract

CONTEXT:

Currently, natural products have been shown to present interesting biological and pharmacological activities and are used as chemotherapeutic agents. Plants have historically been used in treating cancer and are recognized for their ability to produce secondary metabolites. Juglans regia L. (Juglandaceae) has medicinal applications to treat a wide range of diseases such as cancer.

OBJECTIVE:

The current study was designed to evaluate the antiproliferative activity of total extract as well as several fractions from the leaves of J. regia.The total phenolics, flavonoids, and condensed tannins content of these extracts were also determined to obtain further information on the correlation between the contents of phenolic compounds and antiproliferative effects as well as the leaf developmental stages.

MATERIALS AND METHODS:

Antiproliferative activity was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and flow cytometry methods against human oral cancer, breast adenocarcinoma and colon adenocarcinoma cell lines. The total phenolics, flavonoids, and condensed tannins were determined by Folin-Ciocalteu, aluminum chloride and butanol-HCl colorimetric methods.

RESULTS:

Our present study has shown that chloroform fraction has the lowest IC(50) values (0.36-0.81 mg/mL) and also induces cell cycle arrest (G0\G1 phase) after a 24 h treatment. The colorimetric methods showed the highest amount of total phenolics, flavonoids, and condensed tannins in the methanol fraction (120.28 ± 2.32, 59.44 ± 0.87, 227.00 ± 4.91 mg/g of dry weight of extract).

DISCUSSION AND CONCLUSION:

The results obtained herein indicate that walnut chloroform fraction may contain effective compounds which can be used as a chemotherapeutic agent.

http://www.ncbi.nlm.nih.gov/pubmed/22906313

2-Springerplus. 2015; 4: 127. (see attached file).

Published online 2015 Mar 14. doi:  10.1186/s40064-015-0871-4

PMCID: PMC4374083

Anticancer effect of black tea extract in human cancer cell lines

Katarína Koňariková, Miriam Ježovičová, Ján Keresteš, Helena Gbelcová, Zdeňka Ďuračková, and Ingrid Žitňanová

Author information ► Article notes ► Copyright and License information ►

Go to:

Abstract

In this study we investigated effects of natural extract from the black tea Camellia sinensis (BTE) against human colon carcinoma cell line HT-29, human breast carcinoma cell line MCF-7, human alveolar carcinoma cell line A549 and healthy cell line NIH-3T3. We identified concentration range for cytotoxic/antiproliferative effects using MTT assay and the trypan blue assay, gel electrophoresis we employed to determine the type of cell death induced by BTE and DNA damage we determined by comet assay. Different concentrations of the extract (0.00078 - 5 μg/mL) we added to the cultured cells and incubated for 216 h. BTE showed cytotoxic effects against all carcinoma cell lines, however HT-29 and MCF-7 cells were more sensitive than A549. BTE showed no antiproliferative effect against healthy cells NIH-3T3 at tested concentrations. We found no apoptotic cell death in HT-29 and MCF-7 cells after 72 h of incubation in case of single administration of BTE but in case of repetitive administration of BTE (BTE was added to the cells each day) we found apoptotic cell death in HT-29 after 72 h incubation. BTE induced also DNA strand breaks and oxidative damage to DNA in carcinoma cells HT-29 and MCF-7.

Keywords: Camellia sinensis, Black tea, Cancer, Protective effect, Apoptosis

Go to:

Introduction

Black tea has a long history of use dating back to China approximately 5,000 years ago. It is made from the dried leaves of Camellia sinensis, a perennial evergreen shrub formerly known as Thea sinensis. It is native to southeastern Asia. Green tea, black tea, and oolong tea are all derived from the same plant. Black tea results from the oxidation of Camellia sinensis leaves. The chemical components in tea include alkaloids (theobromine, caffeine, theophylline), polyphenols, amino acids, polysaccharides, volatile acids, vitamins, lipids as well as inorganic elements (Liang et al. 2013; Xiaorong et al. 2015; Scoparo et al. 2014). Black tea is used for treating headaches, low blood pressure, preventing heart disease, including atherosclerosis and heart attack, preventing Parkinson’s disease, reducing the risk of stomach and colon cancer, lung, ovarian and breast cancers (Lee and Foo 2013). The aim of our study was to evaluate the potential anticancerogenic effect of the black tea extract (BTE) on different types of carcinoma cell lines.

3- Phytomedicine. 2015 Jan 15;22(1):1-4. doi: 10.1016/j.phymed.2014.09.008. Epub 2014 Oct 22.

Cytotoxic effects of stem bark extracts and pure compounds from Margaritaria discoidea on human ovarian cancer cell lines.

Johnson-Ajinwo OR1, Richardson A1, Li WW2.

Author information

Abstract

Margaritaria discoidea (Baill.) G. L. Webster (Euphorbiaceae) is a well-known medicinal plant in Africa used for the treatment of various diseases. So far, no cytotoxic effects of plant extracts on cancer cell lines have been reported.

AIM OF THE STUDY:

To evaluate the cytotoxicity against human ovarian cancer cells of extracts of M. discoidea and characterize the major bioactive compounds.

METHODS:

Both organic and aqueous extracts of this plant were obtained by maceration. The sulforhodamine B cell proliferation assay was used for evaluation of their cytotoxic activities and the potential bioactive compounds were characterized by gas chromatography-mass spectrometry.

RESULTS:

The organic extract of M. discoidea showed stronger cytotoxicity than the aqueous extract with IC50 values of 14.4±3.0, 14.2±1.2 and 34.7±0.5µg/ml on OVCAR-8, A2780 and cisplatin-resistant A2780cis ovarian cancer cells, respectively. The organic extract was further subjected to bioassay-guided fractionation by partitioning with n-hexane, ethyl acetate, and n-butanol in water. The ethyl acetate fraction was the most potent on the three ovarian cancer cell lines. A GC-MS analysis of trimethylsilyl derivatives of this fraction indicated the presence of phenolic compounds such as gallic acid and the alkaloid securinine. The IC50 values of these two compounds were determined to be in the range of 3-16µM, which indicated that they could contribute to the cytotoxic activity of the extract of M. discoidea.

CONCLUSIONS:

This study has evaluated the cytotoxicity of stem bark extracts of M. discoidea against ovarian cancer cells and provided a basis of further development of this plant for the treatment of ovarian cancer.

Copyright © 2014 Elsevier GmbH. All rights reserved.

KEYWORDS:

GC–MS; Gallic acid; Margaritaria discoidea; Ovarian cancer; Securinine

http://www.ncbi.nlm.nih.gov/pubmed/25636863

4-Nutrients 2015, 7, 2707-2718; doi:10.3390/nu7042707 (see attached file)

nutrients

ISSN 2072-6643

www.mdpi.com/journal/nutrients

Article

Cytotoxic Activity of Piper cubeba Extract in Breast Cancer

Cell Lines

Potchanapond Graidist 1,2,

*, Mananya Martla 1 and Yaowapa Sukpondma 3

Abstract: This study aimed to evaluate the cytotoxicity of a crude extract of Piper cubeba against normal and breast cancer cell lines. To prepare the extract, P. cubeba seeds were ground, soaked in methanol and dichloromethane and isolated by column chromatography. Fractions were tested for cytotoxicity effects on normal fibroblast (L929), normal breast (MCF-12A) and breast cancer cell lines (MCF-7, MDA-MB-468 and MDA-MB-231). The most effective fraction was selected for DNA fragmentation assay to detect apoptotic activity. The results showed that the methanolic crude extract had a higher cytotoxic activity against MDA-MB-468 and MCF-7 than a dichloromethane crude extract. Then, the methanolic crude extract was separated into six fractions, designated A to F. Fraction C was highly

active against breast cancer cell lines with an IC50 value less than 4 μg/mL. Therefore, Fraction C was further separated into seven fractions, CA to CG. The 1

H-NMR profile showed that Fraction CE was long chain hydrocarbons. Moreover, Fraction CE demonstrated the highest activity against MCF-7 cells with an IC50 value of 2.69 ± 0.09 μg/mL and lower cytotoxicity against normal fibroblast L929 cells with an IC50 value of 4.17 ± 0.77 μg/mL. Finally, DNA fragmentation with a ladder pattern characteristic of apoptosis was observed in MCF-7, MDA-MB-468, MDA-MB-231 and L929 cells, but not in MCF-12A cells.

Hoping this will be helpful,

Rafik