Cytotoxic and cytogenetic effects of some extracts of convolvulus arvensis on tumor and transformed cell lines in vitro

العناوين الأخرى

التأثيرات السمية الخلوية و الوراثية لبعض مستخلصات نبات المديد على الخطوط الخلوية السرطانية و المتحولة في الزجاج

مقدم أطروحة جامعية

Hasan, Kawthar Khalaf

مشرف أطروحة جامعية

Sukkar, Dhamya Qasim
al-Asadi, Asad Abd al-Wahid Badr

أعضاء اللجنة

al-Rikabi, Nahi Yusuf Yasin
Nada, Sad Muhammad
Ahmad, Fayizah Abd al-Wahhab
al-Asadi, Iqbal Jasim
Isa, Adnan Badran

الجامعة

جامعة البصرة

الكلية

كلية العلوم

القسم الأكاديمي

قسم علوم الحياة

دولة الجامعة

العراق

الدرجة العلمية

دكتوراه

تاريخ الدرجة العلمية

2010

الملخص الإنجليزي

-Plant plays an important role as a source of many anticancer drugs, thus screening of these plants may gives a new valuable drug with minimal side effects.

For this purpose this study was designed to evaluate the cytotoxic effects of (aqueous and methanolic) crude extracts of leaves,stem and root as well as proteoglycan and glycoside leaves extracts of Convolvulus arvensis on human Rhabdomyosarcoma (RD) tumor cell line and Rat embryo fibroblast (REF-3) cell line in vitro.

The study also included the investigation of the effect of glycoside extract on mitotic index of RD tumor cell line and human lymphocytes as well as damaged cells after the estimation of the cytotoxic concentration 50% (CC50%) of each effective extract.

Eight concentrations of two fold dilution of each extract were prepared and tested on each cell line starting from 10000 μg/ml.

ending with 78.125 μg/ml.

with three replicates for each concentration, the optical density of cell growth read by Elisa reader at 492 nm by used tetrazolium bromide (MTT) The result of in vitro study for RD cell growth assay has shown that the glycoside extract had more cytotoxic inhibition after 24, 48 and 72 hours, the effect was time independent against the proliferation of cells whereas the glycoside extract was more effective at 48 and 72 hours on the proliferation of REF-3.The inhibitory effect on the proliferation of RD started from156 .25 μg /ml.

and 312.5 μg/ml.

on REF-3.

On the other hand glycoside extract had cytotoxicity concentration 50% (CC50%) lower than other extracts on RD tumor cell lines were 1775, 870 and 706 μg/ml in all periods of times with an exception for the stem aqueous and methanolic extracts had less CC50% than glycoside 113.333and 482.5 μg/ml.

respectively after 72 hour only.

Whereas the II CC50% of glycoside was >10000 , 9650 and 7540 μg/ml after 24,48 and 72 hours respectively for REF-3 cell line and more than 10000μg/ml of all other extracts (aqueous, methanolic of leaves, stem and root as well as proteoglycan) for all periods of time.

The root aqueous extract had less cytotoxic effect on the proliferation of RD tumor cell line than other extracts only at 48 and 72 hours .

Whereas it had less cytotoxic effect after all period of times in the REF-3 cell line.

The effect of root aqueous extract on proliferation of both types of cell lines occur only with higher concentration 10000 μg/ml .

The CC50% of the root aqueous extract was more than10000 μg/ml on the proliferation of both types of cell lines.

The study showed that the glycoside had a biphasic effect on RD and REF-3, a stimulatory effect at low concentrations, and an inhibitory effect at high concentrations after 24 and 48 hours.

The proteoglycan, (aqueous and methanolic) extracts of stem had the same biphasic effect on RD tumor cell line for 24h.

The result revealed that RD tumor cell line was more sensitive than REF-3 cell line to all extracts and the optical density values were 0.539 and 0.685 respectively.

The cytogenetic study revealed that the glycoside extract showed a significant effect on the mitotic index of the RD cell line ,since the mitotic index values were 1.93, 3.44 and 6.71% for tested cell line treated with 353, 176.5 and 88.25 μg /ml respectively as compared to mitotic index of the control group 10.34 % .

Structural and numerical chromosomal aberrations were observed in both treated and untreated groups.

The structural aberrations included: Ring chromosome, dicentric chromosome, chromatid break and gap, terminal chromatid addition and terminal chromatid deletion with acentric fragment, III multiredial interchange as well as pulverization.

Numerical chromosomal aberration include polyploidy and aneuploidy.

The results of cytogenetic effect of high concentration 353 μg/ml of glycoside extract on human lymphocytes revealed reducing in mitotic index similar to that positive control group(treated with mitomycin C), whereas the lower concentrations 176.5 and 88.25 μg/ml was less effective than higher concentration but all of them caused less damaged cells(any cell containing one or more chromosomal aberrations) than the positive control and more than negative control.

Some of structural and numerical chromosomal aberrations were observed in treated , positive and negative groups.

Structural aberrations included : Ring chromosome, dicentric chromosome, chromatid break , gap , short arm chromatid deletion, multiredial interchange as well as pulverization.

Numerical chromosomal aberration include polyploidy and aneuploidy.

The study was demonstrated that the glycoside significantly inhibits the treated tumor cell line proliferation and caused apoptosis.

Since the light and fluorescent microscope observation showed morphological characteristic of apoptosis including cell volume shrinking, chromatin condensation and nuclear fragmented were observed after the treatment of RD tumor cell line with different concentration of glycoside.

Beside that the apoptotic effect was confirmed by DNA fragmentation into multiply of low molecular weight fragments 180-200 pb.

as well as fragments with high molecular weight by using of agarose gel electrophoresis when treated with different concentrations.

التخصصات الرئيسية

الأحياء

الموضوعات

• الخلايا السرطانية

عدد الصفحات

194

قائمة المحتويات

• APA
• MLA
• AMA

لغة النص

الإنجليزية

نوع البيانات

رسائل جامعية

رقم السجل

BIM-317100