Here, we established hela cells that overexpress either hsp105. Approximately 60% of hela cells became apoptotic within 4 h as detected by dna fragmentation, terminal deoxynucleotidyltransferasemediated dutpbiotin nick end labeling tunel assay, and electron microscopy. Johns hopkins never patented hela cells, nor did it sell them. Hela cells 1951 she was poor, black and terminally ill with cervical cancer. The fact that hela cells have been used in some very important, groundbreaking medical research is interesting enough, but theres another part of the story and that part is why oprah might be making a movie about hela. Cyt c and other proteins are released to the cytosol apoptosis begins in normal cells, the release of cyt c is blocked by antiapoptotic bcl2 proteins bcl2 stops the aggregation of bh123 for apoptosis to start, bcl2 must be inactivated by binding to a proapoptotic protein e. Hela cell, a cancerous cell belonging to a strain continuously cultured since its isolation in 1951 from a patient suffering from cervical carcinoma. Atg5 and bcl2 provide novel insights into the interplay. A proapoptotic effect of the cdk inhibitor p57kip2 on.
Untreated and sts treated hela and jurkat lysates were prepared in 1x cell extraction buffer ptr and tested the cleaved parp simplesteptm elisa. May 17, 2018 in biology, apoptosis is one of the main types of programmed cell death pcd. Apoptotic volume decrease avd is a characteristic cell shrinkage observed during apoptosis. Although avd has traditionally been attributed to water loss, direct evidence for that is often lacking. Interferon induces the apoptosis of cervical cancer hela. M, is an effective and reliable tool for the induction of apoptosis in hela cells, as reported also in an earlier study from this laboratory. These compounds inhibited cell growth in hela cells at the s phase. Nothing visible, but i felt a couple tiny specs hit me.
Defining characteristics of types i and ii apoptotic cells in. Learn vocabulary, terms, and more with flashcards, games, and other study tools. A read is counted each time someone views a publication summary such as the title, abstract, and list of authors, clicks on a figure, or views or downloads the fulltext. Apoptosis, also called programmed cell death, in biology, a mechanism that allows cells to selfdestruct when stimulated by the appropriate trigger. Rebecca skloot later wrote a popular book on the subject, called the. The socalled hela cells became one of the most important contributions to medical. Figure 1a shows that stp caused a time dependent increase in ht29 apoptosis. For example, treatment with staurosporine, a protein kinase c inhibitor, but not with antifas mab induces apoptosis in hela cells, which are fas. Apoptotic death of a cell and its subsequent phagocytosis by a neighboring cell or by a macrophage allow the organic components of the death cell to be effectively recycled. Apoptosis is a process common to all multicellular organisms. It is commonly used to induce apoptosis in experimental settings, although the mechanism is not fully understood. Molecular mechanism of staurosporineinduced apoptosis in. A proapoptotic effect of the cdk inhibitor p57 kip2 on. One of the cells grown from the cervical cancer of a young africanamerican woman, henrietta lacks.
Use features like bookmarks, note taking and highlighting while reading hela cells of henrietta lacks. Apoptosis can be triggered by mild cellular injury and by various factors internal or external to the cell. The immortal life of henrietta lacks is the story of that discovery and the impact it had on the whole family especially henriettas youngest daughter deborah. The most striking feature of apoptosis is that if a cell undergoes the programmed cell death, the neighboring cells are not at all damaged. Skloot gives an incredible overview of the scientific history of the cells as well as the medical ethics that were developing right along with the growth of hela cells. Staurosporine has long been used in vitro as an initiator of apoptosis in many different cell types, but the mechanism involved remains poorly understood. Farber2 department of pathology, thomas jefferson university, philadelphia, pennsylvania 19107 abstract the role of the mitochondrial permeability transition mpt in the killing of hela cells by staurosporine. Hydrogen peroxide induces apoptosis in hela cells through. It has been found that one way in which staurosporine induces apoptosis is by activating caspase3. Type i cells have been defined to be independent of mitochondria for the induction of fas death receptormediated apoptosis, whereas type ii cells are mitochondriadependent. Vousden, 2 and moshe oren 14 department of chemical immunology, the weizmann institute of science, rehovot 76100, israel. Profilin 1 potentiates apoptosis induced by staurosporine. Apoptotic cells % apoptosis cells numbertotal cells number. Hela cells one book one lcc 201415 the immortal life of.
Induction of the mitochondrial permeability transition. Hela cells cotransfected with pegfp and the indicated plasmid were treated as indicated staurosporine 2. More than 60,000 papers have been written about hela cells 9. Fifty cells per treatment were counted for each concentration point. Apoptosis of hela cells induced by a new targeting photosensitizerbased pdt via a mitochondrial pathway and er stress donghong li,1 lei. Hd phase images provide a qualitative assessment of morphological correlates to apoptosis left. Henrietta lacks had no idea that her cells were taken and used in this way, and neither did her family.
Staurosporine cas 62996741 98% purity ab120056 abcam. The chloride channel inhibitor furosemide prevented intracellular alkalinization, bax translocation, cytochrome c release, and cell. Leonard tolmach the fame of the technique of mitotic collection decided then that we must first clone the cells to avoid any heterogeneity and continuity with the same cell line in years to come. Download it once and read it on your kindle device, pc, phones or tablets. Hela cells of henrietta lacks kindle edition by singh, nishi. Atg5 and bcl2 provide novel insights into the interplay between apoptosis and autophagy.
Buy hela cells of henrietta lacks by nishi singh isbn. There were only a few hela cell lines at that time. May 19, 2017 genetically, hela cells contain parts of henrietta lackss own dna, mutations introduced by the strain or strains of hpv that infected her, as well as uncounted numbers of new mutations. Its been estimated that if you could pile all hela cells ever grown onto a scale, theyd weigh more than 50 million metric tonsas much as a hundred empire state buildings. In this study, we quantified intracellular water in. Spike cell cultures at a staurosporine concentration of 1 m in the cell culture media. Suicidal cells and the immortal cells of henrietta lacks the japan. Today i was working with hela cells in the tissue culture and i think some hela cells splashed onto my chin. We induced apoptosis in murine osteoblast mc3t3e1 cells by exposure to the staurosporine.
The inhibitory effects of daphnoretin on viability and proliferation of hela cells were determined by the mtt assay. Here, we report that selective expression of the third member in this family, p57 kip2, potentiated staurosporineind uced apoptosis. It may be argued that hypertonic stress and hicc activation just delays, rather than prevents, stsinduced cell death in hela cells. M staurosporine was markedly potentiated by cycloheximide. Induction of apoptosis in hela cells with staurosporine produced a rise in the intracellular ph phi. Apoptosis in hela cells is inhibited by proteins produced by the cell. Necrosis was observed at the longest times of recovery as a result of secondary necrosis. Intracellular alkalinization was accompanied by translocation of bax to the mitochondria, cytochromec release, and cell death. A proapoptotic effect of the cdk inhibitor p57 kip2 on staurosporine induced apoptosis in hela cells article in biochemical and biophysical research communications 2963. Rapid apoptosis induced by shiga toxin in hela cells.
There are at least two known processes that may result in the avd. 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. Apoptosis was induced in hela cells treated with staurosporine, a compound know to induce apoptosis by activation. This change in the caspase cascade suggests cdv induces apoptosis via the intrinsic pathway, excluding the need for the initiator caspase8. Hela cell line expresses the human papilloma virus 18, e6 oncoprotein which causes the ubiquitin mediated degradation of p53 protein and is thus p53 deficient. Typical cell suspension concentrations that have been used for this staurosporine induction protocol range from 1 x 105 1 x 106 cells ml.
Right hela nuclight red cells treated with 300 nm staurosporine. The immortal life of henrietta lacks by jasmine akhtar on prezi. These compounds have been found to induce apoptosis in hela cells. I was using the hoodgloves and everything, but i dont think my chair was high enough for the shield on the hood to protect the lower half of my face.
Induction of apoptosis in hela cells by transactivationdeficient p53 ygal haupt, 1 sheldon rowan, 1 eitan shaulian, karen h. At lower concentration, depending on the cell type, staurosporine induces specific cell cycle effects arresting cells either in g 1 or in g 2 phase of the cell. Using these cscs as a model, we tested the effect of doc on the proliferation and differentiation of cscs and demonstrated its inhibitory activity in both processes. Henrietta lacks was a 31yearold mother of five when she died in 1951, but her name and memory live on in the form of a remarkable lineage of continually dividing cells that have achieved, to all intents and purposes, immortality. Imbalance of tumor cells apoptosis regulation is one of the main reasons for the occurrence of tumors. The designation hela is derived from the name of the patient, henrietta lacks. Since we could detect caspase3like activity in the lysate of hela s3 cells treated with etoposide, we tried to image caspase3like protease activity in apoptotic hela s3 cells. To elucidate alterations in mitochondrial function during stsinduced apoptosis in hela. Staurosporineinduced neuronal death was not accompanied by an increase in.
Apr 14, 2017 5 important ways henrietta lacks changed medical science. The apoptotic volume decrease is an upstream event. The kit contains enough primary and secondary antibodies to perform two western. Staurosporine induced neuronal death was accompanied by cell body shrinkage, chromatin condensation, and dna laddering. Vousden, 2 and moshe oren 14 department of chemical immunology.
Apoptosis antibody sampler kit cell signaling technology. Hela was the subject of a book by rebecca skloot, the immortal life of henrietta lacks. Apoptosis, or programmed cell death, is involved in many biological events, including tumorigenesis. On the other hand, activation of the map kinase mapk cascade is also known to play a critical role in apoptosis. Everyday low prices and free delivery on eligible orders. Studies on apoptosis in hela cells via the rosmediated. Hela cells were the first human cell line to be established and have. Aug 15, 2009 indeed, rbac is a very efficient fluorogenic substrate and easily enters the cells where the original photoactive molecule is restored by specific esterases. Based on morphological and biochemical criteria, staurosporine induced apoptosis in a human corneal endothelial cell line has been clearly demonstrated in this study.
George and margaret gey at johns hopkins in baltimore. Cells for kids science book for children singh, nishi on. Intracellular alkalinization was accompanied by translocation of bax to the mitochondria, cytochrome c release, and cell death. It was first introduced by kerr, wyllie, and currie. Knockout studies in mice show that thymocytes are type i and liver cells are type ii. Ijms free fulltext the apoptotic volume decrease is. The percentage of apoptotic cells was calculated as follows. In her 2010 book, the immortal life of henrietta lacks, rebecca skloot documents the histories of both the cell linecalled the hela cell line. Apoptosis in hela cells induced by ssii and evaluated by the aoeb double staining over 6 days. Both apoptosis and necrosis occur under different circumstances and involve different steps. Hela cell apoptosis caused by cdv follows a different mechanism than that in vero cell lines. The apoptotic volume decrease is an upstream event of map kinase activation during staurosporine induced apoptosis in hela cells. Apoptosis definition, pathway, significance and role.
After testing the 2 envelope glycoproteins hemagglutinin h and fusion f, we found that mv h alone was sufficient to induce the apoptosis of hela cells. Pdf apoptosis regulation of hela cells by cwnt5a gene. Open books open minds the immortal life of henrietta lacks by rebecca skloot. Rebecca skloot in her fascinating book, the immortal life of henrietta lacks. This concentration works well for inducing many cell lines including hl60 and jurkat cells when using a 45 hour 37c incubation period. Hsp105 family proteins suppress staurosporineinduced. Full text apoptosis of hela cells induced by a new targeting. Staurosporine induces apoptosis of melanoma through caspasedependent and independent pathways to confirm the role of caspase activation in staurosporine induced apoptosis of melanoma, we treated igr3 and melrm cells with the pancaspase inhibitor, zvadfmk, 1 h before adding staurosporine. Imaging of caspase3 activation in hela cells stimulated with.
Thus, it is suggested that activation of p38 and jnk is a downstream event of avd for the stsinduced apoptosis of hela cells. The patch clamp technique revealed that the k current of the hela cell was inhibited up to 80% when exposed to tea. The book is about henrietta lacks and the immortal cell line, known as hela, that came from lackss cervical cancer cells in 1951. The mitochondrion, as the cell s base for aerobic respiration and site of energy supply, plays an important role in the occurrence of cell apoptosis. Mutations, apoptosis and cancer flashcards quizlet. In this study, we investigated the potential for different components of the measles virus mv to induce apoptosis of hela cells and explored the apoptotic molecular mechanisms. The real science of henrietta lackss immortal cells inverse. As a result, scientific studies about certain cells are actually discussing other. This kind of programmed cell suicide is called apoptosis. Molecules free fulltext laminarin induces apoptosis. Downregulation of atg5 in hela cells reduced cell death and. Mar 08, 2011 the immortal life of henrietta lacks did not require that at all and yet for some reason a large portion of the book is devoted to showing how fair this white and how often they remind us that she is white. Hela ev and hela axin cells were treated with staurosporine sts for indicated time and subjected to immunoblot analysis using the indicated antibodies. A poptosis is one of the most fundamental biological processes in mammals, in which individual cells.
However, apoptosis induced by cdv in hela cells does not involve the initiator protein caspase8. Sep 10, 2012 systems biology of apoptosis summarizes all current achievements in this emerging field. The staurosporine treated hela lysate is designed for use as a western blot positive control for antibodies against apoptosis specific proteins or for experimental treatments to induce apoptosis. Head to for more on henrietta lacks, whose cells advanced scientific. Hela cells open books open minds the immortal life of. A dying cancer cell with stringy protrusions called filopodia, extending to the right, that help cancer cells to migrate away from their original tumors. Mechanisms of staurosporine induced apoptosis in a human.
Dec 02, 2012 hela cells were essential in developing the polio vaccine in 1955 8. Rebecca skloot and her bestseller, the immortal life of henrietta lacks has created a lot of interest in the immortal hela cells that were taken from a 31 year old black woman called henrietta lacks without her familys consent at the johns hopkins hospital at baltimore in 1951. Axin expression reduces staurosporineinduced mitochondria. Staurosporine transiently increased the phosphotransferase activity of cjun nterminal kinase1 jnk1, which in turn may activate the transcriptional activity of activating protein1 ap1. Hela and jurkat cells were treated with 1 m staurosporine sts ab120056 for 4 hours in complete cell culture media to induce apoptosis and cleaved parp protein. The immortal life of henrietta lacks, by rebecca skloot, is the story of rebecca skloots journey in discovering the truth behind hela cells. Pdf the apoptotic volume decrease is an upstream event. M, 5 h, sorted for gfp positivity by flow cytometry, and plated. Moreover, the generation of reactive oxygen species after apoptosis. The apoptosis antibody sampler kit provides an economical means to evaluate the levels of inactive and active caspases. Scanning electron micrograph of an apoptotic hela cell. Many books have been written about the hela henrietta lacks cells and the their effect on medical science. Hela cells by staurosporine1 marco tafani, dmitri a.
Transient expression of sp1 in hela cells slightly increased cell viability after the treatment with staurosporine, which induced apoptosis. Staurosporine induces apoptosis of melanoma by both. In this study, we developed a new strategy to enrich cscs in human cervical carcinoma cells line, hela cells. Significant nuclear damage of hela cells was observed after a comet assay. Axin expression reduces staurosporine induced mitochondriamediated cell death in hela cells. Recent studies done with hela cells providesinformation about the end replication problem. Hela cells are the first line of immortal cells grown in culture. Oct 12, 2017 for decades, immortal cells such as the famous hela cells have been contaminating other cell cultures in the lab. Doxycycline induces apoptosis and inhibits proliferation and. In the present study, we investigated the relationship between the avd induction and the stressresponsive mapk cascade activation during the apoptosis process induced by staurosporine sts in hela cells. Hela cells after pdt underwent a consistent rate of apoptosis peaked at 12 h of recovery post. Hela cells of henrietta lacks by nishi singh, paperback. Figure 1 summarizes the effect of stp on apoptosis of ht29 cells and the relationship to cellular and extracellular gsh and gssg status.
Perform time course studies on your particular cell line to ascertain the optimal staurosporine concentration and exposure time required to achieve good apoptosis. Aug 07, 20 i first came across hela cells as a postdoc in 1972. The term apoptosis is derived from the greek word meaning dropping or falling off. Because none of the synthetic probes was membranepermeable, we used a microinjection technique. In the present study, we have examined the apoptosis inducing potential of staurosporine in cultured melanoma cell lines and dissected the staurosporine induced apoptotic signaling pathway. Blended phasecontrast and redgreen images taken at 20x illustrate homogenous express of red nuclear signal and activation of caspase37 green. Untreated and sts treated hela and jurkat lysates were prepared in 1x cell.
Exposure of murine cortical cell cultures to 30100 nm staurosporine induced concentrationdependent selective neuronal degeneration over the following day. Hemagglutinin protein of measles virus induces apoptosis. Staurosporine induced apoptosis in ht29 cells and the relationship to cellular gsh efflux. Hela cells were the first human cells to be continuously grown in culture. Hela is an immortal cell line used in scientific research. Her book has become a notable science reading for high school children and science. Induction of apoptosis in hela cells with staurosporine produced a rise in the intracellular ph ph i. Mechanistically, the increased pfn1 expression mediated the.
Staurosporineinduced apoptotic water loss is cell and. Autoschizis is a novel type of programmed cell death characterized by exaggerated membrane damage and the progressive loss of cytoplasm through a series of selfexcisions. Approximately 60% of hela cells became apoptotic within 4 h as detected by dna. Apoptosis leads to the elimination of cells via a complex but highly defined cellular programme. Sp1, a type of insect hemocyanin protein, is a femalespecific protein in silkworm and consists of high portion of methionine. Staurosporine is a potent, nonselective inhibitor of protein kinases, including protein kinase c. It is concluded from these experiments that sts, at 4. In this study, hela cells were used as a testing model for the treatment of ifn on cervical cancer.
Regulation of intracellular ph mediates bax activation in. Staurosporine, a microbial alkaloid, is a strong inhibitor of protein kinases. M staurosporine triggers apoptosis in nearly 40% of the cells after 12 hours, rapidly activates caspase3, and induces parp cleavage. Staurosporine induced apoptosis of endothelial cells involves activation of caspase3, and could be a useful model to study strategies of cell death inhibition. In this study, we showed that pfn1 overexpression sensitizes cancer cells to apoptosis through the typical intrinsic apoptotic pathway. Induce apoptosis with staurosporine immunochemistry. The cells were first cultured in february 1951 by drs. Flow cytometry analysis indicated an increased apoptosis rate of the hela cell after exposing to tea. Recently, it has been reported that two members of the cipkip family of cdk inhibitors, p21 cip1 and p27 kip1, are involved in the regulation of apoptosis. Inhibition of drp1dependent mitochondrial fragmentation.
When apoptosis was induced by staurosporine in hela cells, sp2 protein suppressed nuclear fragmentation and apoptotic body formation. Staurosporine is a cell permeable alkaloid isolated from streptomyces staurosporeus exhibiting anticancer activity. The use of hela cells in medical research will be restricted under a new agreement with the family of henrietta lacks, who gave the cells without consent more than 60 years ago. Staurosporineinduced neuronal apoptosis sciencedirect. Jun 23, 2010 h enrietta lacks, a 31yearold mother of five, died of cervical cancer on 4 october 1951. Cellplayer 96well kinetic caspase37 apoptosis assay. Induction of apoptosis in hela cells by transactivation. This equates to a 1 l spike of the 1 mm staurosporine stock per ml of the cell culture suspension. Hypertonicityinduced cation channels rescue cells from.
897 1106 1229 178 214 378 364 889 172 659 442 742 78 947 255 615 663 827 342 1202 369 1213 121 112 877 611 635 1508 611 1459 160 77 432 540 1012 917 1129 427 1333 1367 593 103 1193 273