Browsing by Author "Bulimo, Wallace"

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    Antiproliferative Activity, c-Myc and FGFR1 Gene Expression Profiles and Safety of Annona muricata Fruit Extract on Rhabdomyosarcoma and BALB/c Mice
    (Journal of Complementary and Alternative Medical Research, 2021) Chikwana, Naomi; Maina, Esther N.; Gavamukulya, Yahaya; Bulimo, Wallace; Wamunyokoli, Fred
    Rhabdomyosarcoma is an aggressive solid tumour of skeletal muscles origin whose current treatment is associated with high expenses, severe side effects, drug resistance and tumour regrowth. There is a need to develop safer and more effective chemotherapeutic agents. Annona muricata is one of the widely used plants in treating various diseases due to its reported effectiveness. However, there is a dearth of scientific information regarding the efficacy of Annona muricata on rhabdomyosarcoma and its safety. This study aimed to evaluate the effects of Annona muricata ethanolic fruit extract on the antiproliferative activity and gene expression in RD cell line, including its biosafety in BALB/c mice. Materials and Methods: The resazurin metabolic assay was used to assess the antiproliferative and cytotoxic activities of Annona muricata ethanolic fruit extract on RD and Vero cells. Quantitative real-time polymerase chain reaction was used to assess the gene expression profiles on c-Myc and FGFR1 genes. To evaluate the safety of the Annona muricata ethanolic fruit extract, an acute oral toxicity study was conducted on BALB/c mice. Results: Annona muricata ethanolic fruit extract significantly inhibited the growth of RD cells in a concentration and time-dependent manner while being highly selective on the Vero cells (selectivity index of 6.10 at 72h) compared to a reference cancer drug, doxorubicin (Selectivity index of 1.38 at 72hr). The c-Myc and FGFR1 genes were under expressed in RD cells treated with Annona muricata ethanolic fruit extract with (3.4 and 6.1 fold), respectively, compared to untreated cells. Acute oral toxicity studies revealed no significant difference (p ≥ 0.05) between the treated mice and the control group, indicating the safety of the fruit extract. Conclusion: Annona muricata ethanolic fruit extract can serve as effective and safe anticancer agents against rhabdomyosarcoma and further develop into standard drugs. Non-human primate studies need to be undertaken to step towards the clinical utilization of the Annona muricata ethanolic fruit extract in the management of rhabdomyosarcoma.
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    Dermaseptin B2’s Anti-Proliferative Activity and down Regulation of Anti-Proliferative, Angiogenic and Metastatic Genes in Rhabdomyosarcoma RD Cells in Vitro
    (Advances in Bioscience and Biotechnology, 2021) Abdille, Ahmed A.; Kimani, Josephine; Wamunyokoli, Fred; Bulimo, Wallace; Gavamukulya, Yahaya; Maina, Esther N.
    Rhabdomyosarcoma (RMS) is the most prevalent soft tissue sarcoma in children, representing approximately 50% of pediatric sarcomas and can develop in any part of the body though more frequently at the extremities. Aim: Evaluating the in vitro anti-proliferative activity of Dermaseptin B2 on Rhabdomyosarcoma RD (CCL-136TM) cells and its effect on the expression of MYC, FGFR1, NOTCH1, and CXCR7 genes involve in processes including proliferation, angiogenesis and metastasis. Methods: RD cells were grown in Dulbecco’s Modified Eagle’s Medium supplemented with 10% Fetal Bovine Serum. Exponentially growing cells were treated with Dermaseptin B2 and Antiproliferative activity was assayed using the resazurin and migration assays at three time-points. In order to determine the gene expression profiles of MYC, NOTCH1, FGFR1 and CXCR7, total RNA was extracted from the cells and q-RT-PCR was performed with β-Actin as reference gene. Results: Dermaseptin B2 inhibited the proliferation of RD cells in a time and concentration dependent manner as with IC50 values of 7.679 μM, 7.235 μM, 5.993 μM. The 2-dimentional wound healing assay showed inhibition of migration and motility of the RD cells at time-points of 6, 24, 48 and 72-hours with the greatest inhibition observed at 72-hours. Dermaseptin B2 downregulated the target MYC (fc; 1.5013, 1.5185, 2.4144), CXCR7 (fc; 2.8818, 4.4430, 3.9924), FGFR1 (fc; 2.3515, 2.0809, 2.2543), NOTCH1 (fc; 2.4667, 4.6274, 4.3352) genes for the three-time points respectively. NOTCH1 and CXCR7 showed higher fold changes with respect to β-Actin than MYC and FGFR1. Conclusion: The results of this study indicate that Dermaseptin B2 is a target molecule for signaling pathways including PI3K/AKT, RTK and NOTCH pathways that could affect the transcription of these genes and overall inhibition of cancer progression. Further studies are needed to give a better understanding of the detailed mechanisms of action as well as the effects of the Dermaseptin B2 peptide in vivo.
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    Sub-acute and sub-chronic toxicity assessment of the antimicrobial peptide Dermaseptin B2 on biochemical, haematological and histopathological parameters in BALB/c mice and Albino Wistar rats
    (Heliyon, 2022) Abdille, Ahmed A.; Kitimu, Shedrack Reuben; Ndubi, Mark M.; Kimani, Josephine; Maina, Esther N.; Bulimo, Wallace; Gavamukulya, Yahaya; Wamunyokoli, Fred
    Dermaseptins (Drs) are peptides found in the skin secretions of a variety of Hylid frogs, particularly those belonging to the Agalychnis and Phyllomedusa families. Dermaseptin B2 (Drs B2), an amphipathic, α-helical polypeptide was reported as the most active of the Dermaseptin B family. We have previously shown that Drs B2 has strong anti-proliferative activities against RD cells in vitro and thus required further evaluations for future medical applications. Aim: The aim the study was to evaluate the 14-day sub-acute and 90-day sub-chronic toxicities Drs B2 in vivo. Materials and Methods: BALB/c mice were treated with increasing concentrations of 5–25 mg/kg of Drs B2. Rats were treated with 2, 4 and 10-fold concentrations of the calculated LD50 of Drs B2 following OECD recommendations. At the end of the experimentation periods, the animals were sacrificed and dissected to collect blood and selected organs for analysis of any effects caused by Drs B2 treatment on the biochemical, haematological, and histological parameters. Results: The 14-day sub-acute toxicity tests did not cause significant alteration in the biochemical, hematological and histological parameters. The 90-day sub-chronic toxicity study showed lower ALT and AST than control at doses 1.9 mg/kg and 4.6 mg/kg, respectively. Their haematology results also showed higher platelet count than the controls but the differences were not statistically significant. Histological analysis showed increased megakaryocytes in the spleen for both the mice and the rats. Conclusion: The results of this study indicate that short term treatment of Drs B2 could be safe to the animals, however, long-term treatment can have mild effects on the liver parameters and cause an inflammatory response in the spleen.