Cytotoxic Effects of Hydro-Alcoholic Extract of the Leaf of Elaeagnus angustifolia in Hepatocellular Carcinoma Cell Line (HepG2)

Background: Cancer is one of the most complicated diseases with various treatments, which each has its special side effects. So, pharmaceutical companies are intended to develop new drugs with minimum side effects. Objectives: The current study aimed to investigate the cytotoxicity effects and the redox potential of alcoholic extract of Oleaster leaf on liver carcinoma cell line (HepG2). Methods: Oleaster leaves were collected from Qazvin (Iran), and the alcoholic extract of the plant leaves was prepared. HepG2 cells were cultured in DMEM medium and treated with 50, 100, 200, 400, and 600 μg/mL of the extract. The cytotoxicity effect of the extract was evaluated using the MTT and the Neutral Red assays. Redox potential in HepG2 cells was assessed using NO, catalase, and GSH tests. The expression of Bax and bcl-2 genes in HepG2 cells was evaluated for apoptosis analysis. Results: The results showed that the extract could significantly (P < 0.001) reduce the viability of HepG2 cells. Also, the extract significantly increased the amount of released NO, catalase activity, and GSH concentration. RT-PCR results showed that Oleaster leaf extract significantly change the expression of bax and bcl-2. Conclusions: The results showed that the leaves of the Oleaster plant contain compounds with cytotoxicity properties, so it can be considered as a potent candidate for liver cancer treatment.

Isolation of cancer stem-like cells from hepatocellular carcinoma cell line HepG2 by methods of magnetic-activated cell sorting, spheroid culture, and anti-tumor drug-resistant selection: A primary evaluation

Introduction: Recently reported data have suggested that only a small subset of cancer cells possess the capability to initiate malignancies. These observations were based on investigation of cells within the primary tumors displaying a distinct surface marker pattern. CD133 marker is a putative hematopoietic and neuronal stem cell marker, which is also considered to be a tumorigenic marker in brain, prostate and liver. Recent studies have shown that a small population of CD133-positive cells, indeed, exists in human hepatocellular carcinoma (HCC) cell lines and primary HCC tissues. This study was aimed at isolating the cancer stem-like cells from hepatocellular carcinoma cell line HepG2 using three different methods: magnetic-activated cell sorting (MACS), spheroid culture (SC), and anti-tumor drug (ATD) resistant selection. Methods: HepG2 hepatocellular carcinoma cells were expanded to yield enough cells that could be used to isolate cancer stem-like cells by these three methods. For MACS, cancer stem-like cells were sorted using anti-CD133 monoclonal antibody. For the second method, cancer stem-like cells were enriched by selection of anti-tumor drug resistance property. Lastly, for the third method, three-dimensional (3D) culture was used to enrich for the cancer stem-like cells. The cells obtained by the three methods were expanded to obtain an adequate number of cells for confirmation of CD133 expression. Results: The expression of CD133+ cells in the three methods was found to be different. In the MACS method, the expanded CD133+ sorted cells cultured through 2 passages only contained 0.40 % CD133+ cells. In the 3D spheroid cell culture, of the population of cells there were 38.39 % that were CD133+ cells. Lastly, in the anti-tumor drug (doxorubicin at 150 nM) resistant selection, 66.22 % were CD133+ cells. Conclusion: This study shows that isolation of HepG2 derived CD133+ population by culture with doxorubicin (150 nM) yields the highest efficiency and purity of the 3 methods studied.

Facile synthesis and antiproliferative activity of new 3-cyanopyridines

Background Pyridines have been reported to possess various pharmacological activities. Results Sodium 3-oxo-3-(2-oxo-2H-chromen-3-yl)prop-1-en-1-olate (2) and sodium 3-oxo-3-(3-oxo-3H-benzo[f]chromen-2-yl)prop-1-en-1-olate (7) were prepared and reacted with 2-cyano-N’-(1-aryl(heteryl)ethylidene)acetohydrazides 3a–d to produce 2-oxo-1,2-dihydropyridine-3-carbonitrile derivatives 5a–d and 9a–d, respectively, in good yields. Also, 3a–d reacted with sodium (2-oxocyclopentylidene)methanolate (11a) or sodium (2-oxocyclohexylidene) methanolate (11b) to yield 2-oxo-tetrahydro-1H-cyclopenta[b]pyridine-3-carbonitriles 13a–d and 2-oxo-hexahydroquinoline-3-carbonitriles 13e–h, respectively. The mechanisms that account for the formation of the products are discussed. Additionally, the structures of all the newly synthesized products are confirmed, based on elemental analysis and spectral data. Several of the newly synthesized compounds are evaluated for their antitumor activity against HEPG2 and their structure activity relationship (SAR) was studied. Conclusions The results revealed that the pyridine derivatives 5c and 5d (IC50 = 1.46, 7.08 µM, respectively) have promising antitumor activity against liver carcinoma cell line (HEPG2), compared to the reference drug, doxorubicin.