Cell death in HeLa cells upon imperatorin and cisplatin treatment

There is growing evidence that commonly applied chemotherapy regimens can be improved by introducingnew, specific, active and low side-effect drugs, or by combining substances to obtain the required clinicaleffect. The aim of the present study was to investigate the effects of imperatorin and cisplatin, applied separatelyor in combination, on apoptosis, necrosis and autophagy induction in the human cervical carcinoma cell line(HeLa). Imperatorin appeared to be a potent autophagy inducer, rather than a necrotic or apoptotic one. Incontrast, cisplatin induced mainly apoptosis and necrosis after 6 h and 24 h, while longer incubation resultedonly in necrosis induction. When HeLa cells were incubated with both drugs, autophagy appeared most frequently,although to a smaller extent than that observed after imperatorin administered alone. At the molecularlevel, autophagy was correlated with the presence of the cleaved form of microtubule-associated protein 1 lightchain LC3 — LC3II. It was also accompanied by the inhibition of heat shock proteins Hsp27 and Hsp72 expression.Our results indicate that imperatorin alone, or in combination with cisplatin, is mainly an autopahgy inducerin HeLa cells.There is growing evidence that commonly applied chemotherapy regimens can be improved by introducingnew, specific, active and low side-effect drugs, or by combining substances to obtain the required clinicaleffect. The aim of the present study was to investigate the effects of imperatorin and cisplatin, applied separatelyor in combination, on apoptosis, necrosis and autophagy induction in the human cervical carcinoma cell line(HeLa). Imperatorin appeared to be a potent autophagy inducer, rather than a necrotic or apoptotic one. Incontrast, cisplatin induced mainly apoptosis and necrosis after 6 h and 24 h, while longer incubation resultedonly in necrosis induction. When HeLa cells were incubated with both drugs, autophagy appeared most frequently,although to a smaller extent than that observed after imperatorin administered alone. At the molecularlevel, autophagy was correlated with the presence of the cleaved form of microtubule-associated protein 1 lightchain LC3 — LC3II. It was also accompanied by the inhibition of heat shock proteins Hsp27 and Hsp72 expression.Our results indicate that imperatorin alone, or in combination with cisplatin, is mainly an autopahgy inducerin HeLa cells

Hepatocyte growth factor (HGF), heat shock proteins (HSPs) and multidrug resistance protein (MRP) expression in co-culture of colon tumor spheroids with normal cells after incubation with interleukin-1<img src='/image/spc_char/beta.gif' border=0> (IL-1<img src='/image/spc_char/beta.gif' border=0>) and/or camptothecin (CPT-11)

354-364Tumor chemoresistance and metastasis are some of the most important problems in colon cancer therapy. In the present study, co-cultures of human colon carcinoma cell spheroids, obtained from different grades of tumor, with human colon epithelium, myofibroblast and endothelial cell monolayers were performed. The purpose of these co-cultures was to reflect, in in vitro conditions, different stages of colon tumor development. In order to investigate the invasive capacities of the tumor cells and their resistance to chemotherapy, HGF, HSP27, HSP72 and MRP levels were analyzed after incubation of the co-cultures with IL-1 and irinotecan (CPT-11) added as single agents or in combination. Myofibroblasts produced significantly higher amounts of HGF than epithelial cells. Tumor cells released trace amounts of this molecule. In co-cultures, IL-1 induced HGF release, while CPT-11 alone or combined with IL-1decreased HGF secretion. An immunoblotting analysis followed by densitometry revealed that the combination of IL-1 plus CPT-11 added to the co-cultures led to a decrease in HSPs and MRP levels. In conclusion, direct and paracrine interactions of colon tumor cell spheroids with normal cells and exogenously added CPT-11 change HSP27, HSP72 and MRP expression in comparison to monocultures. IL-1 and CPT-11, dependent on whether they are added separately or jointly, differentially modulate HGF expression in monocultures of colon tumor spheroids or normal cells and their co-cultures

Essential Oil from Arnica Montana L. Achenes: Chemical Characteristics and Anticancer Activity

Mountain arnica Arnica montana L. is a source of several metabolite classes with diverse biological activities. The chemical composition of essential oil and its major volatile components in arnica may vary depending on the geographical region, environmental factors, and plant organ. The objective of this study was to characterize the chemical composition of essential oil derived from A. montana achenes and to investigate its effect on induction of apoptosis and autophagy in human anaplastic astrocytoma MOGGCCM and glioblastoma multiforme T98G cell lines. The chemical composition of essential oil extracted from the achenes was examined with the use of Gas Chromatography&ndash;Mass Spectrometry GC-MS. Only 16 components of the essential oil obtained from the achenes of 3-year-old plants and 18 components in the essential oil obtained from the achenes of 4-year-old plants constituted ca. 94.14% and 96.38% of the total EO content, respectively. The main components in the EO from the arnica achenes were 2,5-dimethoxy-p-cymene (39.54 and 44.65%), cumene (13.24 and 10.71%), thymol methyl ether (8.66 and 8.63%), 2,6-diisopropylanisole (8.55 and 8.41%), decanal (7.31 and 6.28%), and 1,2,2,3-tetramethylcyclopent-3-enol (4.33 and 2.94%) in the 3- and 4-year-old plants, respectively. The essential oils were found to exert an anticancer effect by induction of cell death in anaplastic astrocytoma and glioblastoma multiforme cells. The induction of apoptosis at a level of 25.7&ndash;32.7% facilitates the use of this secondary metabolite in further studies focused on the development of glioma therapy in the future. Probably, this component plays a key role in the anticancer activity against the MOGGCCM and T98G cell lines. The present study is the first report on the composition and anticancer activities of essential oil from A. montana achenes, and further studies are required to explore its potential for future medicinal purposes

Quercetin suppresses heat shock-induced nuclear translocation of Hsp72

The effect of quercetin and heat shock on the Hsp72 level and distribution in HeLa cells was studied by Western blotting, indirect immunofluorescence and immunogold electron microscopy. In control cells and after quercetin treatment, Hsp72 was located both in the cytoplasm and in the nucleus in comparable amounts. After hyperthermia, the level of nuclear Hsp72 raised dramatically. Expression of Hsp72 in cytoplasm was also higher but not to such extent as that observed in the nucleus. Preincubation of heated cells with quercetin inhibited strong Hsp72 expression observed after hyperthermia and changed the intracellular Hsp72 distribution. The cytoplasmic level of protein exceeded the nuclear one, especially around the nucleus, where the coat of Hsp72 was noticed. Observations indicating that quercetin was present around and in the nuclear envelope suggested an involvement of this drug in the inhibition of nuclear translocation. Our results indicate that pro-apoptotic activity of quercetin may be correlated not only with the inhibition of Hsp72 expression but also with suppression of its migration to the nucleus

Cell death in HeLa cells upon imperatorin and cisplatin treatment

There is growing evidence that commonly applied chemotherapy regimens can be improved by introducing&lt;br /&gt;new, specific, active and low side-effect drugs, or by combining substances to obtain the required clinical&lt;br /&gt;effect. The aim of the present study was to investigate the effects of imperatorin and cisplatin, applied separately&lt;br /&gt;or in combination, on apoptosis, necrosis and autophagy induction in the human cervical carcinoma cell line&lt;br /&gt;(HeLa). Imperatorin appeared to be a potent autophagy inducer, rather than a necrotic or apoptotic one. In&lt;br /&gt;contrast, cisplatin induced mainly apoptosis and necrosis after 6 h and 24 h, while longer incubation resulted&lt;br /&gt;only in necrosis induction. When HeLa cells were incubated with both drugs, autophagy appeared most frequently,&lt;br /&gt;although to a smaller extent than that observed after imperatorin administered alone. At the molecular&lt;br /&gt;level, autophagy was correlated with the presence of the cleaved form of microtubule-associated protein 1 light&lt;br /&gt;chain LC3 — LC3II. It was also accompanied by the inhibition of heat shock proteins Hsp27 and Hsp72 expression.&lt;br /&gt;Our results indicate that imperatorin alone, or in combination with cisplatin, is mainly an autopahgy inducer&lt;br /&gt;in HeLa cells.There is growing evidence that commonly applied chemotherapy regimens can be improved by introducing&lt;br /&gt;new, specific, active and low side-effect drugs, or by combining substances to obtain the required clinical&lt;br /&gt;effect. The aim of the present study was to investigate the effects of imperatorin and cisplatin, applied separately&lt;br /&gt;or in combination, on apoptosis, necrosis and autophagy induction in the human cervical carcinoma cell line&lt;br /&gt;(HeLa). Imperatorin appeared to be a potent autophagy inducer, rather than a necrotic or apoptotic one. In&lt;br /&gt;contrast, cisplatin induced mainly apoptosis and necrosis after 6 h and 24 h, while longer incubation resulted&lt;br /&gt;only in necrosis induction. When HeLa cells were incubated with both drugs, autophagy appeared most frequently,&lt;br /&gt;although to a smaller extent than that observed after imperatorin administered alone. At the molecular&lt;br /&gt;level, autophagy was correlated with the presence of the cleaved form of microtubule-associated protein 1 light&lt;br /&gt;chain LC3 — LC3II. It was also accompanied by the inhibition of heat shock proteins Hsp27 and Hsp72 expression.&lt;br /&gt;Our results indicate that imperatorin alone, or in combination with cisplatin, is mainly an autopahgy inducer&lt;br /&gt;in HeLa cells

Antiglioma Potential of Coumarins Combined with Sorafenib

Coumarins, which occur naturally in the plant kingdom, are diverse class of secondary metabolites. With their antiproliferative, chemopreventive and antiangiogenetic properties, they can be used in the treatment of cancer. Their therapeutic potential depends on the type and location of the attachment of substituents to the ring. Therefore, the aim of our study was to investigate the effect of simple coumarins (osthole, umbelliferone, esculin, and 4-hydroxycoumarin) combined with sorafenib (specific inhibitor of Raf (Rapidly Accelerated Fibrosarcoma) kinase) in programmed death induction in human glioblastoma multiforme (T98G) and anaplastic astrocytoma (MOGGCCM) cells lines. Osthole and umbelliferone were isolated from fruits: Mutellina purpurea L. and Heracleum leskowii L., respectively, while esculin and 4-hydroxycoumarin were purchased from Sigma Aldrich (St. Louis, MO, USA). Apoptosis, autophagy and necrosis were identified microscopically after straining with specific fluorochromes. The level of caspase 3, Beclin 1, PI3K (Phosphoinositide 3-kinase), and Raf kinases were estimated by immunoblotting. Transfection with specific siRNA (small interfering RNA) was used to block Bcl-2 (B-cell lymphoma 2), Raf, and PI3K expression. Cell migration was tested with the wound healing assay. The present study has shown that all the coumarins eliminated the MOGGCCM and T98G tumor cells mainly via apoptosis and, to a lesser extent, via autophagy. Osthole, which has an isoprenyl moiety, was shown to be the most effective compound. Sorafenib did not change the proapoptotic activity of this coumarin; however, it reduced the level of autophagy. At the molecular level, the induction of apoptosis was associated with a decrease in the expression of PI3K and Raf kinases, whereas an increase in the level of Beclin 1 was observed in the case of autophagy. Inhibition of the expression of this protein by specific siRNA eliminated autophagy. Moreover, the blocking of the expression of Bcl-2 and PI3K significantly increased the level of apoptosis. Osthole and sorafenib successfully inhibited the migration of the MOGGCCM and T98G cells

The Dual Blockade of the TIGIT and PD-1/PD-L1 Pathway as a New Hope for Ovarian Cancer Patients

The prognosis for ovarian cancer (OC) patients is poor and the five-year survival rate is only 47%. Immune checkpoints (ICPs) appear to be the potential targets in up-and-coming OC treatment. However, the response of OC patients to immunotherapy based on programmed cell death pathway (PD-1/PD-L1) inhibitors totals only 6–15%. The promising approach is a combined therapy, including other ICPs such as the T-cell immunoglobulin and ITIM domain/CD155/DNAX accessory molecule-1 (TIGIT/CD155/DNAM-1) axis. Preclinical studies in a murine model of colorectal cancer showed that the dual blockade of PD-1/PD-L1 and TIGIT led to remission in the whole studied group vs. the regression of the tumors with the blockade of a single pathway. The approach stimulates the effector activity of T cells and NK cells, and redirects the immune system activity against the tumor. The understanding of the synergistic action of the TIGIT and PD-1/PD-L1 blockade is, however, poor. Thus, the aim of this review is to summarize the current knowledge about the mode of action of the dual TIGIT and PD-1/PD-L1 blockade and its potential benefits for OC patients. Considering the positive impact of this combined therapy in malignancies, including lung and colorectal cancer, it appears to be a promising approach in OC treatment

Mathematical Prostate Cancer Evolution: Effect of Immunotherapy Based on Controlled Vaccination Strategy

Basic immunology research over several decades has led to an improved understanding of tumour recognition by components of the immune system and mechanism of tumour evasion from immune detection. These findings have ultimately led to creating antitumour immunotherapies in patients with different kind of cancer including prostate cancer. The increasing number of reports confirms that immune-based therapies have clinical benefit in patients with prostate cancer with potentially less toxicity in comparison with traditional systemic treatments including surgical resection, chemotherapy, or radiotherapy in various forms. This review focuses on the possibility of modulation of the optimal immunotherapy based on vaccination strategy adopted to individual patients in order to increase quality and quantity of their life