Thesis (M.Sc)-Cairo University, 2025.

Bibliography: pages 87-103.

Capparis spinosa L., an edible medicinal plant, is rich in flavonoids. Production of
secondary metabolites preferentially uses amino acids as precursors using in vitro culture
techniques. This study aimed to evaluate the effect of amino acids as elicitors on
morphological characterization,non-antioxidant compounds accumilation, antioxidant
enzymes, and anticancer activity Capparis spinosa was in vitro cultured on MS media
containing three amino acids included; tryptophan, glutamine or phenylalanine at 100,
200, and 300 mg/L and compared with free amino acid culture (as control). After four
weeks morphological characteristics were determined, some phytochemicals panel and
rutin were evaluated by spectrophotometric and HPLC, respectively. The hepatocellular
carcinoma cell line HepG2 was utilized to validate the anti cancer activity and in silico
studies. The results showed that using colorex 15% resulted in 91.67% of
disinfection.Applying 6-Benzylaminopurine(BAP) at 0.8 mg/L in MS media gave the
highest shootlet length(4.03cm) ,but kinetin at 04 mg/L gave the highest shootlet number
(6.11shoot/explant) . the most significant result (100%) to callus induction was achieved
with MS medium supplemented with 0.5 mg/L 2,4-D for leaves.The MS media
supplemented with 200 mg/L of tryptophan produced the most significant results, with an
average shootlet number of 34.67. The explants grown on MS media supplemented with
100 mg/L of phenylalanine showed the greatest shootlet length (6.83 cm). Moreover, the
explant grown on MS media with 200 mg/L of tryptophan had the largest number of
leaves (16.32).Regarding fresh weight, the control group recorded the highest fresh plant
weight 3.50 g, while the highest fresh weight of callus was 4.7 g under the treatment with
tryptophan at 100 mg/L or glutamine at 300 mg/L. .The HPLC analysis of rutin for the
callus treated with glutamine at 100 mg/L or phenylalanine 200 mg/L (the highest
treatments of flavonoids) scored 88.2 and 64.3 µg/g, respectively. The toxicity of callus
extract treated with glutamine at 100 mg/L was more toxic for cancer cells (HepG2)
followed by phenylalanine at 200 mg/L at IC50 356.65 and 371.05 µg/mL, respectively.
Additionally, in molecular docking, rutin effectively reduced the activity in the HepG2
promoter. It downregulated the mRNA expression of HepG2-responsive genes of liver
cancer cells, reducing its capacity to form spheroids and blocking the HepG2 signaling
pathway. These findings suggest that amino acid elicitors can improve the metabolism of
Capparis spinosa L. and could be used to produce high amounts of rutin, which has
antitumor activity against HepG2. Preventing oxidative damage and inducing apoptosis
may be the mechanisms responsible for the anti-tumor effect. Rutin from Capparis
spinosa presents a promising candidate for further in silico and in vitro anticancer
research, particularly through molecular docking studies to explore its interactions with
key oncogenic targets.

هدفت الدراسة إلى اختبار تأثير ثلاثة أحماض أمينية (تربتوفان، جلوتامين، فينيل ألانين) على نمو نبات الكباريس وإنتاج مركباته الثانوية ونشاطه المضاد للسرطان. أثبت محلول الكلوروكس 15% أعلى كفاءة تعقيم، بينما حقق BAP أعلى طول للساق والكينيتين أعلى عدد للأفرع. وأظهر التربتوفان 200 ملجم/لتر أفضل نمو من حيث عدد الأفرع، في حين أعطى الفينيل ألانين 100 ملجم/لتر أطول الأفرع. سجّل الجلوتامين 100 ملجم/لتر والفينيل ألانين 200 ملجم/لتر أعلى محتوى من الروتين. وأظهر الكالس المعالج بالجلوتامين 100 ملجم/لتر أقوى تأثير مضاد لخلايا سرطان الكبد HepG2. كما بيّن الربط الجزيئي قدرة الروتين على خفض التعبير الجيني للخلايا السرطانية وإيقاف مسار إشاراتها، مما يجعله مرشحًا واعدًا لأبحاث علاج السرطان.

Issues also as CD.

Text in English and abstract in Arabic & English.