The Effects of Colchicum baytopiorum on Regulatory Genes of Apoptotic and Autophagic Cell Death in HeLa Cells.

BACKGROUND: The natural products have increasing important for the development of anticancer agents. Colchicum baytopiorum C.D. Brickell (C. baytopiorum), an endemic species for Turkey, contains colchicine and its derivatives. Stimulation of apoptotic and autophagy-mediated cell deaths are effective strategy for anticancer therapies. AIM: The aim of the study is to determine the role of the extract on both apoptotic and autophagic cell death in HeLa cell line. METHODS: The cell viability of C. baytopiorum (0.1 mg/ml) was determined by MTT assay. Active caspase-3 and t-Bid expressions were evaluated by immunohistochemical method. The mRNA expression of apoptotic regulatory genes (Bcl-xL, Bid, Bad, PUMA, NOXA, Caspase-3, -8, -9, Fas, FADD, TRADD, TRAF2, TNF, TNFR1), autophagic cell death related genes (Atg5-12, Beclin-1, DAPK), and also both autophagic and apoptotic cell death regulatory genes (Bif-1 and BNIP-3) were investigated by qRT-PCR. RESULTS: We determined that the expressions of both apoptotic and autophagic regulatory genes were significantly increased in the treatment group compared to control group. Also, we showed that C. baytopiorum crude extract induces the cross-connection between apoptotic and autophagic cell deaths in HeLa cells. CONCLUSION: We suggested that this endemic plant extract seems to be a new promising therapeutic approach in cancer.

In vitro anticancer and cytotoxic activities of some plant extracts on HeLa and Vero cell lines.

PURPOSE: The aim of our study was to evaluate the effect of in vitro anticancer and cytotoxic activity of the methanolic extracts of 14 medicinal plants, 8 of which are endemic species in Anatolia, against the human HeLa cervical cancer cell line and to compare to the normal African green monkey kidney epithelial cell line (Vero) using the MTT colorimetric assay. METHODS: Values for cytotoxicity measured by MTT assay were expressed as the concentration that causes 50% decrease in cell viability (IC50, µg/mL). The degree of selectivity of the compounds can be expressed by its selectivity index (SI) value. High SI value (>2) of a compound gives the selective toxicity against cancer cells (SI = IC50 normal cell/IC50 cancer cell). RESULTS: Dose-dependent studies revealed IC50 of 293 mg/mL and >1000 mg/mL for Cotinus coggygria Scop., IC50 of 265 µg/mL and >1000 mg/mL for Rosa damascena Miller, IC50 of 2 µg/mL and 454 mg/mL for Colchicum sanguicolle K.M. Perss, IC50 of 427 µg/mL and >1000 µg/mL for Centaurea antiochia Boiss. var. praealta (Boiss & Bal) Wagenitz on the HeLa cells and the Vero cells, respectively. Four plants showed significant SI values which were 227 for Colchicum sanguicolle K.M. Perss (endemic species), >3.8 for Rosa damascena Miller, >3.4 for Cotinus coggygria Scop. and >2.3 for Centaurea antiochia Boiss. var. praealta (Boiss & Bal)Wagenitz (endemic species). CONCLUSION: According to our study, 4 methanolic extracts of 14 tested plants exhibit greater activity on the HeLa cell line and little activity on the Vero cell line, meaning that these plants can be evaluated for potential promising anticancer activity.

Screening for antitumor activity of various plant extracts on HeLa and C 4-I cell lines.

PURPOSE: Cancer is a long process that leads the organism to death and is associated with the normal cells acquiring the ability to divide permanently. Nowadays, the use of natural products in cancer therapy has a great importance. In addition, working with plants that are endemic to Turkey and determining the biological activities of these plant extracts, is extremely important due to the potential for new drug development. There is no comparative study available in the literature on the antitumor effects of Colchicum sanguicolle, a new found species of the genus Colchicum in Turkey, Crateagus microphylla, of the genus Crateagus and Centaurea antiochia of the genus Centaurea. In this study, we tried to demonstrate the antitumor effect of these plant extracts on HeLa and C 4-1 cells. METHODS: Five different doses (0.001, 0.01, 0.05, 0.25 and 0.5 mg/ml) of the three plant types were prepared and applied for 24, 48 and 72 hrs on the cervical cancer derived cell lines. Subsequently, the growth rate was evaluated with the mitochondrial dehydrogenase enzyme method. RESULTS: Colchicum sanguicolle extracts showed the most effective antitumor activity. For the Colchicum sanguicolle extract, the IC50 dose for HeLa cells was 0.01 mg/ml at 48 hrs, while for the C-4 I cells it was 0.001 mg/ml at 48 hrs. These results showed that C-4 I cells were more sensitive to the Colchicum sanguicolle extracts. Conclus?on: The results of from this study regarding the antitumor effect of plant extracts of endemic varieties of Turkey may have an important place in design and development of anticancer drugs and would make contributions to other studies to be conducted in this area.

Antibacterial effect of Kenger gum on mutans streptococci and its cytotoxic effect on the 3T3 fibroblast cell line.

PURPOSE: To evaluate the antibacterial effect of Kenger gum on mutans streptococci (in vivo) and Streptococcus mutans (in vitro) and its cytotoxic effect on the 3T3 fibroblast cell line. MATERIALS AND METHODS: In vitro antibacterial activity of Kenger gum extracts against S.mutans was determined by the disk-diffusion method. The broth dilution method was used to determine the minimum inhibitory concentration (MIC). The cytotoxic effect on 3T3 fibroblast cells at different time intervals was determined using cell culture and viability assays. Clinical studies were then performed on 20 healthy adult subjects, where a sugar-free chewing gum was used as a control. To determine the MS counts, oral rinse samples were taken before chewing as well as 30 and 60 min after 15 min of chewing. Repeated-measures ANOVA was used to compare the bacteria level in the oral rinse samples between the two chewing gums. The Least Significant Difference test was used for adjustment for multiple comparisons. RESULTS: The MIC of the acetone extract of Kenger gum was 30 µg/ml. The acetone extract of Kenger gum possessed moderate antiproliferative properties against the non-tumorigenic cell line 3T3. A statistically significant decrease was observed for both chewing gums at 30 and 60 min. The decrease continued at 60 min after chewing Kenger gum, while the values for control gum tended to approach the baseline after 60 min. CONCLUSION: This preliminary study showed that Kenger gum had particular and prolonged antibacterial activity against S. mutans and salivary mutans streptococci.