Uv-induced apoptosis in resistant hela cells book

It has also been shown that the breast cancer tumor suppressor, brca1, is phosphorylated on ser1423 and ser1524 by the ataxia telangiectasia mutatedrelated kinase atr after uv damage. Restoration of uv sensitivity in uv resistant hela cells. Molecular mechanisms of uvinduced apoptosis the number of sc 26. Role and regulation of p53 during an ultraviolet radiation. Ultraviolet b uvb irradiationinduced apoptosis in various. In the present work, we investigated the extent of oxidative stress, dna damage, apoptosis and expression of apoptosis. In human epithelial hela cells stimulated with antifas antibody or sts, the avd induction was found to precede activation of caspase8 and caspase9 and to be resistant to pancaspase blockers. Dec 27, 2012 morphological observations showed that low doses of uv induced apoptosis, whereas higher doses triggered both apoptosis and necrosis. The phosphorylation cascade of kinases in the pi3kaktmtor signaling pathway was significantly inhibited by meth exposure, as were proteins downstream of mtorc1, such as p70s6k, rps6, 4ebp1 and eif4e. Ligand binding characteristics of a glycosylphosphatidyl inositol membraneanchored hela cell folate receptor. Meth inhibited proliferation of u118 mg cells and induced apoptosis and autophagy. Apoptosis of hela cells induced by a new targeting.

No induction of apoptosis was found in hacat since this line is defect of p53 we are not tested apoptosis in hfb. These microrna are considered as oncogenes which improve proliferation, inhibits apoptosis and induce tumor angiogenesis. Apoptosis of damaged keratinocytes antagonizes uv induced malignant transformation and thus is a protective pathway. Tnf has been shown to modulate proliferation, differentiation, and apoptotic or necrotic cell death in a number of different cell types.

Handbook of neurochemistry and molecular neurobiology, pp. Nongenetic origins of celltocell variability in trail. The uvtreated hela lysate is designed for use as a western blot positive control when studying uvinduced dna damage andor apoptosis. Some agents have no effect on a cell line, whereas other agents might induce necrosis, rather than apoptosis, or even induce different apoptosis pathways. Cell elimination by apoptosis occurs when uv induced cell damage which cannot be repaired by the.

Pias1 rnai significantly reduced the levels of pias1 protein in hela cells. Critical roles of the pi3kaktmtor signaling pathway in. Distinct signaling pathways of microtubule inhibitors. Whereas the extent of uv induced activation of apoptosis executioners, including dna fragmentation factor, and caspase3 were reduced in the uv resistant cells compared with those apparent in the. The cell line was found to be remarkably durable and prolific. In this study, we investigated the induction of apoptosis in hela cells and its role in acquired uvresistance. Journal of huazhong university of science and technology medical sciences 2009, 29, 625630. The percentage of the apoptotic cell after exposure to 50. Untreated cells were grown under standard cell culture conditions for hela cells. Because with an equitoxic dose of uv light the same frequency of proliferating cells underwent apoptosis figure 2c, it is reasonable to conclude that apoptosis is a main cause of uvcinduced cytotoxicity, both in nerdeficient 271 and wildtype cells. Nucleocytoplasmic transport in apoptosis cell death. To further understand the role of pias1 in uvinduced apoptosis, we treated hela cells with pias1 rnai for 48 hours, and then the cells were uvirradiated fig. Sustained suppression of fas ligand expression in cisplatin resistant. The membrane receptor fas was induced toassembly, and its immediate.

Uvinduced ubiquitylation of xpc protein mediated by uvddb. Sw cells with ectopic brg1 expression regain active swisnf and become significantly more resistant to uv radiation. Cells were grown in dmem supplemented with 10% fetal calf serum, penicillin 100 uml, glutamine 2 mm, and streptomycin 100. The term apoptosis refers to a series of specific morphological changes associated with cell death. Uv induced apoptosis could you tell please me any protocol for induction of apoptosis with uv light for example for hek 293t or hrptc humen primery renal proximal tubal cells. Trailr4 promotes tumor growth and resistance to apoptosis in cervical carcinoma hela cells through akt academic article trail induced apoptosis is enhanced by heat shock protein 70 expression academic article targeting bcl2 to enhance vulnerability to therapy in estrogen receptorpositive breast cancer academic article.

Biochemical events lead to characteristic cell changes and death. Natural products as anticancer molecules have drawn the attention of researchers and have resulted in the development of many successful anticancer drugs, which include camptothecins, epipodophyllotoxins, vinca alkaloids, and taxanes. Pretreatment of murine keratinocytes with ir before uv reduced the apoptotic. The line was derived from cervical cancer cells taken on february 8, 1951 from henrietta lacks, a 31yearold africanamerican mother of 5, who died of cancer on october 4, 1951.

Mitochondrial continuous intracellular networkstructures. Uvinduced apoptosis in resistant hela cells springerlink. The sumo ligase pias1 regulates uvinduced apoptosis by. Culture conditions, transfection, microinjection, and timelapse analysis. Previous studies have shown that mouse embryonic stem cells deficient for caspase3 are resistant to uvinduced apoptosis and that the active form of caspase3 translocates to the nucleus. The translocator protein tspo is an outer mitochondrial membrane protein that is widely used as a biomarker of neuroinflammation, being markedly upregulated in activated microglia in a range of brain pathologies. Tumor cells expressing rcc2yfp are resistant to apoptosis. Mar 20, 20 a prototype of uv induced apoptosis of keratinocytes is the formation of epidermal sunburn cells that are destined to apoptosis after sun exposure. Involvement of exo1b in dna damageinduced apoptosis. These results strongly suggest that both cpds and 64pps contribute to uvinduced apoptosis in nerdeficient cells, while in nerproficient cells, cpds are the only lesions responsible for uvkilling, probably due to the rapid repair of 64pps by ner. Apoptosisinduced proliferation in uvirradiated human. These data indicated that dna damage is crucially involved in uvinduced apoptosis. Ginsenosides are major pharmaceutical components in ginseng, cytotoxicity of ginsenoside rh2, which exists in ginseng extract with trace concentrations, on cancer cells has been known.

However, the regulators of the choice between cell death and survival signaling have not been completely elucidated. Thanks to the elimination by apoptosis of cells harboring genetic mutations, the host is able to prevent further incorporation of damaged andor mutated genes in subsequent clonal cell expansion. An antiapoptotic protein human survivin is a direct. These changes include blebbing, cell shrinkage, nuclear fragmentation, chromatin condensation, chromosomal dna fragmentation, and global vague mrna decay.

Apoptosis of hela cells induced by a new targeting photosensitizerbased pdt via a mitochondrial pathway and er stress donghong li,1 lei li,2 pengxi li,1 yi li,3 xiangyun chen1 1state key laboratory of trauma, burn and combined injury, the second department of research institute of surgery, 2the first department of research institute of surgery, 3cancer center, daping hospital, third military. Apparent alterations in the early stage of excision repair of uv induced dna damages in a hela mutant cell line that is resistant to genotoxic stresses chuck c. C hela cells were transfected with control or exo1 sirna before. Jul 25, 2005 it is important, however, to emphasize that p53 is not the sole mediator of uv induced apoptosis as p53negative cells can seamlessly undergo uvb induced apoptosis, strongly suggesting the presence of additional mechanisms of cell death. Mar, 2008 our focus has been on determining the role of the jnk pathway in cell death induced by microtubule inhibitors.

Several potential lead molecules such as camptothecin cpt and its analogues have been isolated from plants with anticancer effect. We show that sw cells synchronized at the g 1 s border progress into s phase. Hence, we investigated the interplay between ir and uv with regard to uv induced apoptosis. Oxidatively generated damage to cellular dna by uvb and uva. Temporal relationship between cytochrome c release and mitochondrial swelling during uvinduced apoptosis in living hela cells.

In this setting, it is frequently combined with uv radiation, primarily for tanning purposes. Apoptosis in hce cells was induced by uv light irradiation 312nm, 4. The xeroderma pigmentosum group c xpc protein complex plays a key role in recognizing dna damage throughout the genome for mammalian nucleotide excision repair ner. Recently, apoptosis genetically programmed cell death induced by uv hasbeen documented in some cell culture models. Caspase3 mediated cleavage of brca1 during uvinduced. To assess the temporal effect of genotoxic agentinduced apoptosis on the integrity of brca1, hl60 cells a wellstudied model system for apoptosis. Apoptosis was measured using the muse annexin v and dead cell assay kit. Restoration of uv sensitivity in uvresistant hela cells. Dna repair in cells sensitive and resistant to cis. Frontiers therapeutic properties of bioactive compounds. Cyclobutane pyrimidine dimers are the most frequent photoproducts generated in dna after uv irradiation, and induce several metabolic responses, such as cell cycle arrest, rna transcription inhibition and apoptosis. Viruses can induce apoptosis of infected cells either directly, to assist virus dissemination, or by inadvertently triggering cellular sensors that initiate cell death. Western blot analysis with a rcc2specific antibody showed both rcc2yfp and endogenous rcc2 in these cells.

This work reports the involvement of damaged dnabinding protein 2 ddb2, a component involved in the genomic repair of uv damage, in the cross resistance of cisplatinselected cell lines to death ligandmediated apoptosis. Despite its extensive use as a target in molecular imaging studies, the exact cellular functions of this protein remain in question. Caspases play an essential role during apoptotic cell death. Caspase1 activity is required for uvbinduced apoptosis of. Uv induces apoptosis, resulting in the generation of chemokines, which recruit more effector memory t cells and plasmacytoid dcs in uvinduced cutaneous les lesions. Apoptosis is implemented by death machinery that involves the caspase family of proteins, under the condition of a variety of stresses. Whereas the extent of uvinduced activation of apoptosis executioners, including dna fragmentation factor, and caspase3 were reduced in the uv resistant cells compared with those apparent in the.

Uvb, which is an oxidant and proapoptotic agent widely demonstrated in keratinocytes, melanocytes and epidermal cells 30 32, appeared also useful to study apoptotic behavior in other cell cultures in. Daphnoretin is a bicoumarin compound isolated from a natural product, wikstroemia indica, which has been used to treat many diseases. These mirna are contextspecific and they have different roles depending on where they are. Cells were treated with uvc light for 1 minute, then cultured for 4 hours before collecting for lysates. Ligand binding characteristics of a glycosylphosphatidyl inositol membraneanchored hela cell folate receptor epitoperelated to human milk folate binding protein.

Effect of cell confluence on ultraviolet light apoptotic. Chao, shanglang huang mutation research letters 1993 303 1, 1927. Induction of apoptosis in hela cells by transactivation. How to induce early apoptosis in cells without using. Some caspases are also required in the immune system for the maturation of lymphocytes.

Functional defects in mitochondria are involved in the induction of cell death in cancer cells. Ultraviolet radiationinduced interleukin 6 release in hela. Uvinduced apoptosis in cell lines was characterized by morphological changes as well as dna fragmentation into unit multiples of 200 bp, which. Different drugs or stimuli can have different effects on the same cell line or tissue. Cell proliferation and dna breaks are involved in ultraviolet. We should note that the frequency of necrosis was not significantly enhanced in 271 cells. Restoration of uv sensitivity in uvresistant hela cells by. Role of g1 phase in the uvinduced apoptosis of el4 mouse lymphoma cells. Jan 01, 2014 caspases are essential in cells for apoptosis, or programmed cell death, in development and most other stages of adult life, and have been termed executioner proteins for their roles in the cell. Hela cells were resistant to trailinduced apoptosis. Here, we report that human adenocarcinoma cells mcf7 survive uvinduced dna damage by heat shock protein 27. It is the oldest and most commonly used human cell line.

Ultraviolet radiationinduced apoptosis in keratinocytes. A second clinical example featuring uvinduced apoptosis is the response of psoriasis to uv phototherapy. B dnabinding activity in parental me cells and resistant meb, mer, and merb cells. Ros generation was measured by using 5and 6 chloromethyl27dichlorodihydrofluorescein diacetate, acetyl ester. Cpds and 64pps play different roles in uvinduced cell. Since the apoptosis program in tumor cells plays a critical role in the chemotherapyinduced tumor cell killing, it is suggested that the defect in the signaling pathway of apoptosis could cause a new form of multidrug resistance in tumor cells. Taking the antitumor activity of daphnoretin as a starting point, the present study aimed to test the proapoptotic effect of daphnoretin and its underlying mechanism in hela cells. Some studies have shown that exercise accelerates this process. One such signal with widespread pleiotropic actions is the cytokine tumor necrosis factor. Ethanolic extract of propolis eep enhances the apoptosis. I either get viable cells or dead cells and nothing in between. Regulation of ionizing radiationinduced apoptosis by. Clinical standardization of these products is hindered by chemical variability depending on honeybee and botanical sources, but different molecules have been isolated and pharmacologically characterized. Brca1 phosphorylation regulates caspase3 activation in uv.

In microorganisms, noise in gene expression gives rise to cell to cell variability in protein concentrations 1,2,3,4,5,6,7. Crossresistance to death ligandinduced apoptosis in. Molecular mechanisms of uvinduced apoptosis and its effects. Some methods such as treatment with kinase inhibitors, exposure to ultraviolet uv light or xray irradiation are more. Rcc2 overexpression in tumor cells alters apoptosis and. Transcriptional activation of p53 target genes, due to dna damage, causes either apoptosis or survival by cell cycle arrest and dna repair. However, the significance of apoptosis in uvinduced cytotoxicity and resistance is uncertain.

Our study demonstrated that eep and its components significantly sensitize to trail induced death in cancer cells. Recent studies showed that genetic disruption of jnk1 alleles alone was sufficient to render immortalized mouse fibroblasts resistant to uv induced apoptosis 37. Finally they have tissuesspecific mirna expression. Uvb, which is an oxidant and proapoptotic agent widely demonstrated in keratinocytes, melanocytes and epidermal cells 30 32, appeared also useful to study apoptotic behavior in other cell cultures in vitro. I am trying to induce early apoptosis in pc3m cells and k562 cells using heat induction.

Melanocytes are resistant to uv induced apoptosis suggesting that p53 activity is somehow blocked nonfunctional p53, a state shared with melanoma cells, which are resistant to conventional modes of chemotherapy that aim to stimulate p53dependent apoptosis. This is notable in skin cells, to prevent melanoma. A hela cells were treated with 40 jm 2 uvc and lysates were taken at the indicated timepoints for immunoblotting with the indicated antibodies. Apoptosis, or cellular suicide, is important for normal development and tissue homeostasis, but too much or too little apoptosis can also cause disease1,2. It is unknown if the activation of these peptidases 22 is part of a global response to uv radiation or is only observed in cells undergoing apoptosis. Hacat cells died by mitotic catastrophe without uniform induction of apoptosis. Molecular mechanisms of uvinduced apoptosis and its. Stable transfection of the resistant cells with ddb2 antisense cdna resulted in marked depletion of ddb2 protein and restored cellular sensitivity to uvinduced apoptosis. This implicates that cyclobutane pyrimidine dimers cause uvinduced apoptosis in human cells. Could you tell please me any protocol for induction of apoptosis with uv light for example for hek 293t or hrptc humen primery renal proximal tubal cells.

Jun 28, 2017 honeybees produce honey, royal jelly, propolis, bee venom, bee pollen, and beeswax, which potentially benefit to humans due to the bioactives in them. Fasmediated apoptosis in jurkat cells is suppressed in the preg2m phase. Restoration of uv sensitivity in uv resistant hela cells by antisensemediated depletion of damaged dnabinding protein 2 ddb2. My method in brief is suspending 250 000 500 000 cells per eppendorf microfuge tube in 100 ul of rpmi. Mpt of hela cells was measured by the incorporation of. These observations further suggest that ddb2 functions in the repair of uv induced dna damage, and that the intracellular concentration of this protein is an important determinant of acquired uv resistance. Recently, apoptosis genetically programmed cell death induced by uv has been documented in some cell culture models. After two postdoctoral positions in the usa in the emerging field of cell cycle regulation, he was recruited to establish his own independent research at the queensland institute of medical research, and then recruited to the diamantina institute in 2002. Nuclear extract from hela cells stimulated with tnf.

B hela cells were treated as in a, cells were harvested at the indicated times and analysed for annexin v staining via facs. Similar observations were made in opposums following photoreactivation 27. Restoration of uv sensitivity in uva resistant hela cells. Uvbinduced apoptosis is the programmed cell death of cells that become damaged by ultraviolet rays. The resultsindicate that uv induces apoptosis by direct activation of apoptoticproteins in hela and resistant cells. The latter radical that has been found to inhibit propagative lipid peroxidation by scavenging transient peroxyl radicals 93 was found to protect uva. Overexpression of ddb2 has also been shown to potentiate global genomic repair in hamster cells. Photorepair prevents ultravioletinduced apoptosis in. Chronic myeloid leukemia cml cells express the active bcrabl1 protein, which has been targeted by imatinib in cml therapy, but resistance to this drug is an emerging problem. Increased activity of cell surface peptidases in hela. In thisstudy, we investigated the induction of apoptosis in hela cells and itsrole in acquired uv resistance. Morphological observations showed that low doses of uv induced apoptosis, whereas higher doses triggered both apoptosis and necrosis. Interestingly, cells derived from ddb2knockout mice are resistant to uvinduced apoptosis, exhibiting a delayed activation of caspase on uvirradiation itoh et al.

The cisplatin resistant cell line hr3 exhibits enhanced expression of ddb2 and cross resistance to uvinduced activation of apoptosis and caspases. The morphological changes of the cells were analyzed by fluorescence microscopy right. Molecular biomarkers of dna damage in diffuse largecell. In order to investigate the role of these lesions in uvinduced apoptosis in human cells, we generated hela cells expressing a marsupial cpdphotolyase gene gene phr. However, the molecular mechanisms underlying uvb induced apoptosis of keratinocytes are only poorly understood van laethem et al. Thus, it is concluded that the avd induction is an early event independent of the mitochondrial apoptotic signaling pathway and initiator caspase. Cells expressing hpv16 e6 were resistant to a uv induced delay at 4 jm 2, and their movement into sphase was delayed, although not completely inhibited, at a uv dose of 8 jm 2.

In fact, in cultured normal keratinocytes, p53 acts most likely as an antiapoptotic protein since these. Hsp27 protects adenocarcinoma cells from uvinduced. Sensitivity to uv light correlates well with dramatic uv induced apoptosis in sw cells, but not in sw cells expressing brg1. Uv induced apoptosis uv induced apoptosis is an adequate physiological reaction of a cell damaged by uv radiation uvr in a sufficiently large lethal dose and it prevents the disordered destruction of uv damaged cells by help necrosis. The silencing of gsta44 in human osteoarthritic chondrocytes increases hne induced cell death. Uvinduced apoptosis in resistant hela cells bioscience.

The process of programmed cell death may occur through the mechanisms of apoptosis. Apoptosis is a physiological process, that promotes the active suicide of cells. Apr 29, 2005 nucleocytoplasmic transport in apoptosis. The impact of ir on uv induced carcinogenesis is still unclear. In the present study, depletion of ddb2 from uv resistant hela cells, which exhibit enhanced dna repair, increased the sensitivity of the cells to uv induced apoptosis. Cells free fulltext the translocator protein tspo in. Necrotic effect versus apoptotic nature of camptothecin in. However, the significance ofapoptosis in uv induced cytotoxicity and resistance is uncertain. Infrared radiation confers resistance to uvinduced apoptosis. Bcrabl1 induces endogenous oxidative stress promoting genomic instability and imatinib resistance. Cells have the capability of responding to a multitude of signals that it encounters in its extracellular environment. In contrast to uv induced apoptosis, uv mediated il6 release by hela cells was not affected at all by accelerated removal of pyrimidine dimers.

Multiple mechanisms of apoptosis induced by 20sprotopanaxadiol appd in glioma cells. Apoptosis resistance in tumor cells pubmed central pmc. In this study, we examined the effect of uvb radiation on hela cells, and to elucidate the mechanisms involved in activating csp in cells undergoing apoptosis. Uv induces immunosuppression which may play a crucial role in skin cancer development 6. Caspasedependent regulation of histone deacetylase 4 nuclear. This phenotype is similar to the resistance of mismatch repair deficient cells to cisplatininduced apoptosis, as discussed above. Exposure of both sensitive and resistant hela cells to uv induced a dosedependent increase in the percentage of apoptotic cells in both ddb2 knockdown cell lines. Effects of livin gene rna interference on apoptosis of cervical cancer hela cells and enhanced sensitivity to cisplatin. Dna damage and the regulation of cell fate, p 845862. Cancer is an uncontrolled growth and division of cells, leading to significant morbidity and mortality and economic burden to the society. In mammalian cells, protein levels also vary 8,9,10 and individual cells. Nov 27, 2008 infrared radiation ir is increasingly used for wellness purposes.