الفهرس 
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Abstract
Hepatocellular carcinoma is one of the most causes of cancer mortality in Egypt and worldwide that needs exploratory research in cellular signaling pathways. Radiotherapy and chemotherapy treatments cause a variety of undesirable side effects threatening human life. Hence, the main objective of this thesis is to study the development of other therapeutic anticancer alternatives that are more effective and less hazardous to our health.
For that Bidens pilosa and Trianthema portulacastrum were chosen to assess their anticancer properties according to the following plan:
1. Estimation of the anticancer properties of fresh leaves DMSO extracts for both plants against HepG2 cells.
2. Determination of the cell viability and cytotoxic concentration using MTT assay.
3. Quantification of the proliferation and autophagy-related genes by qRT-PCR.
4. Investigation of proinflammatory cytokines using the ELISA.
These extracts were evaluated for their cytotoxic activity and potential concentration (CC50) by MTT assay in HepG2 cells and normal cells. Accordingly, the cell viability rate of treated HepG2 cells was interrupted at a concentration of 0.5 mg/mL of B. pilosa DMSO extract. Hence, the number of survived cells was highly significantly down-regulated in HepG2 cells treated with this extract, whereas it revealed a non-significant change upon treatment with T. portulacastrum. Moreover, normal hepatocytes treated with B. pilosa extract showed non-significant difference in cell morphology when compared to DMSO-treated and non-treated cells. Interestingly, the plant agent disrupted the cancer cells at a low concentration with no substantial cytotoxic impact on the normal cells, according to the CC50 of its extract on the normal cells.
The expressions of Raf-1, MEK-1, LC3B, and Atg12 genes in the HepG2 cell lines were quantified using qRT-PCR, and the cellular total RNA. Also, the released IL-1α and IL-1β were quantified using ELISA assay. The study demonstrated that B. pilosa extract highly significantly reduced the relative expression of proliferation (Raf-1, MEK-1), and autophagy (LC3B, and Atg12) related-genes. T. portulacastrum extract, on the other hand, led to a significantly -reduced level of MEK-1 and a non-significant difference in the Raf-1, LC3B, and Atg12 genes.
Also, B. pilosa extract elevated the IL-1α and IL-1β levels that induced the apoptosis process. Collectively, these data revealed that this extract effectively blocked the Raf/MEK/ERK proliferation signaling pathway and caused a potent inhibition of autophagy process by regulating Raf-1, MEK-1, LC3B and Atg12 respectively, as a potential synergistic effect in liver cancer cells. Additionally, it increased the IL-1α and IL-1β activity and induced the P53 as an apoptotic signaling pathway and eventually, it significantly reduces carcinogenesis. Interestingly, On the basis of the present investigation, we propose that the leaf extract of B. pilosa can be a good candidate for the search of efficient environmentally friendly natural bioactive agents and pharmaceutically important compounds.
Figure 21 Schematic representation of the whole role of Bidens pilosa extract as an anticancer agent for hepatocellularcarcinoma