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M C Joiner

Publications and source records attributed to M C Joiner.

At least 19 recordsLinked to original sources
PubMed2001

Low-dose hypersensitivity: current status and possible mechanisms.

PURPOSE: To retain cell viability, mammalian cells can increase damage repair in response to excessive radiation-induced injury. The adaptive response to small radiation doses is an example of this induced resistance and has been studied for many years, particularly in human lymphocytes. This review focuses on another manifestation of actively increased resistance that is of potential interest for developing improved radiotherapy, specifically the phenomenon in which cells die from excessive sensitivity to small single doses of ionizing radiation but remain more resistant (per unit dose) to larger single doses. In this paper, we propose possible mechanisms to explain this phenomenon based on our data accumulated over the last decade and a review of the literature. CONCLUSION: Typically, most cell lines exhibit hyper-radiosensitivity (HRS) to very low radiation doses (<10 cGy) that is not predicted by back-extrapolating the cell survival response from higher doses. As the dose is increased above about 30 cGy, there is increased radioresistance (IRR) until at doses beyond about 1 Gy, radioresistance is maximal, and the cell survival follows the usual downward-bending curve with increasing dose. The precise operational and activational mechanism of the process is still unclear, but we propose two hypotheses. The greater amount of injury produced by larger doses either (1) is above a putative damage-sensing threshold for triggering faster or more efficient DNA repair or (2) causes changes in DNA structure or organization that facilitates constitutive repair. In both scenarios, this enhanced repair ability is decreased again on a similar time scale to the rate of removal of DNA damage.

Adaptation, Physiological↗
PubMed2001

The translational research chain: is it delivering the goods?

PURPOSE: To address whether the translational research chain has influenced clinical practice in radiation oncology. METHODS AND MATERIALS: Merits and limitations of the various steps of the translational chain, i.e., in vitro studies, animal experiments, biomathematical modeling, Phase I and II trials, and randomized Phase III trials are briefly reviewed. The process and value of translational research in radiation oncology are addressed using dose fractionation and the time factor in tumors as examples. RESULTS: The examples show that translational research may indeed change clinical practice in radiation oncology. However, it takes several decades and considerable efforts to define and test new strategies. The "translational process" is by no means unidirectional but a continuing multiway dialog among basic scientists, applied scientists, clinical scientists, and clinical oncologists. CONCLUSION: Translational research works in radiation oncology, and it is difficult to conceive a better alternative for future improvement of therapy. The slow speed of the translational process indicates that there is a need for improving the various steps of the translational network and the interaction as a whole. Massive investments in one part of the network are likely to be at least partly wasted unless the other links are strengthened as well.

Animals↗
PubMed2001

Low-dose hypersensitivity after fractionated low-dose irradiation in vitro.

PURPOSE: It was demonstrated previously that some radioresistant tumour cell lines respond to decreasing single, low radiation doses by becoming increasingly radiosensitive. This paper reports the response of four radioresistant human glioma cell lines to multiple low-dose radiation exposures given at various intervals. Three of the cell lines (T98G, U87, A7) were proven already to show low-dose hyper-radiosensitivity (HRS) after single low doses; the fourth, U373, does not show HRS after acute doses. MATERIALS AND METHODS: Clonogenic cell-survival measurements were made in vitro using the Dynamic Microscopic Image Processing Scanner (DMIPS) or Cell Sorter (CS) following exposure to 240kVp X-rays one or more times. RESULTS: A consistent, time-dependent hypersensitive response to a second, or subsequent, dose was observed in the cell lines that demonstrated HRS. This time-dependent change in radiosensitivity did not occur in the radioresistant cell line that did not show HRS (U373). In one cell line that demonstrated strong HRS, T98G, a similar time-dependent hypersensitive response was also seen when the cells were irradiated whilst held in the G1-phase of the cell cycle. In this same cell line, significantly increased cell kill was demonstrated when three very low doses (0.4 Gy) were given per day, 4 h apart, for 5 days, compared with the same total dose given as once-daily 1.2Gy fractions. CONCLUSIONS: These data demonstrate the possibility that a multipledose per day, low-dose per fraction regimen, termed 'ultrafractionation', could produce increased tumour cell kill in radioresistant tumours compared with the same total dose given as conventional-sized 2 Gy fractions.

Astrocytoma↗
PubMed2001

Automated counting of mammalian cell colonies.

Investigating the effect of low-dose radiation exposure on cells using assays of colony-forming ability requires large cell samples to maintain statistical accuracy. Manually counting the resulting colonies is a laborious task in which consistent objectivity is hard to achieve. This is true especially with some mammalian cell lines which form poorly defined or 'fuzzy' colonies, typified by glioma or fibroblast cell lines. A computer-vision-based automated colony counter is presented in this paper. It utilizes novel imaging and image-processing methods involving a modified form of the Hough transform. The automated counter is able to identify less-discrete cell colonies typical of these cell lines. The results of automated colony counting are compared with those from four manual (human) colony counts for the cell lines HT29, A172, U118 and IN1265. The results from the automated counts fall well within the distribution of the manual counts for all four cell lines with respect to surviving fraction (SF) versus dose curves, SF values at 2 Gy (SF2) and total area under the SF curve (Dbar). From the variation in the counts, it is shown that the automated counts are generally more consistent than the manual counts.

Automation↗
PubMed2001

A purpose-built iodine-125 irradiation plaque for low dose rate low energy irradiation of cell lines in vitro.

The phenomenon of hyper-radiosensitivity (HRS) to very low acute single doses of radiation has been demonstrated in several cell lines in vitro and in vivo, and has been studied in theory and in practice. The theory suggests a similar hypersensitivity when cells are continuously exposed to radiation at very low dose rates. These low dose rates are used when radioactive seed (iodine-125 or palladium-103) implants of the prostate are used as an alternative to surgery or external beam radiotherapy. To investigate the radiobiology of hypersensitivity of this type on various cell lines in vitro, an iodine-125 seed irradiator has been designed and built for safe use in the Gray Laboratory. In practice, the calculated dose rate has been used for consistency. Discrepancies between calculated and measured dose rates are discussed.

Brachytherapy↗
PubMed2000

Expression of proteins coincident with inducible radioprotection in human lung epithelial cells.

Human lung epithelial cells and many other cell lines are hypersensitive to low doses of ionizing radiation (<0.2 Gy). However, above a threshold dose of 0.4-0.6 Gy, an induced radioprotective response is triggered that protects cells at higher radiation doses. At 4 h, when maximal induced radioprotection is seen in these cells after low-dose priming, the two-dimensional gel protein expression pattern in 0.5-Gy-exposed cells is subtly altered, with seven proteins being 2- to 5-fold down-regulated and one being 2-fold up-regulated. They include: (a) the protein kinase C inhibitor 1, or histidine triad nucleotide-binding motif (HINT) protein; (b) substrates for protein kinase C activity including the chloride intracellular channel protein 1; and (c) a cytoskeletal protein degraded during apoptosis. In addition, a lung cancer-specific protein that binds to both telomeres and nascent mRNA molecules is down-regulated, as is interleukin 1alpha. Therefore, at least in human lung epithelial cells, radioprotection may be the result of signaling pathway switching, which results in the removal of damaged cells and the preparation for enhanced general transcription in surviving cells during a period in which cell proliferation is repressed. This combination of events may be cell-type-specific and may have implications for the protection of normal lung tissue during unavoidable radiation exposure such as in radiotherapy.

Dose-Response Relationship, Radiation↗
PubMed2000

Increased repair and cell survival in cells treated with DIR1 antisense oligonucleotides: implications for induced radioresistance.

PURPOSE: To determine whether repression of a recently isolated, X-ray-responsive gene, DIR1, using antisense oligonucleotides could affect clonogenic cell survival and repair of DNA strand breaks and have a possible role in the mechanism underlying the phenomenon of 'induced radioresistance' (IRR). MATERIALS AND METHODS: Three cell lines, V79, RT112 and UM-UC-3, which are known to exhibit low-dose hypersensitivity (HRS) and induced radioresistance (IRR), and the radiosensitive cell line ATBIVA, were transfected with antisense oligonucleotides directed towards the DIR1 gene. Scrambled oligonucleotides were used as controls. DNA single-strand break (ssb) repair, using the alkaline comet assay, and cell survival using a standard clonogenic assay was measured after exposure to X-rays. RESULTS: Following treatment with 4Gy X-rays, the V79, RT112 and UM-UC-3 cell lines all exhibited significantly increased rates of ssb repair after transfection with DIR1 antisense oligonucleotides compared with cells transfected with scrambled oligonucleotides. They also demonstrated significantly enhanced survival after exposure to 2 Gy X-rays; the radiosensitive ATBIVA cells did not show these effects. CONCLUSIONS: Repression of the DIR1 gene product leads to an increase in the rate of repair and cell survival in three radioresistant cells lines but not in the radiosensitive ATBIVA cell line. Because DIR1 is repressed by X-rays in the dose range where IRR is observed, it may represent a candidate gene involved in the IRR phenomenon.

Animals↗
PubMed2000

Modification of survival by DNA repair modifiers: a probable explanation for the phenomenon of increased radioresistance.

PURPOSE: The low dose (<1 Gy) survival curve of mammalian cells is characterized by a region of hypersensitivity (HRS) followed by increased resistance (IRR). Above 1 Gy, the survival response can be described with a smooth downward-bending curve. Indirect evidence has indicated that the IRR response might reflect an induced radioresistance triggered by DNA damage. The work reported here provides experimental evidence that consolidates this hypothesis. MATERIALS AND METHODS: Clonogenic survival of V79-379A cells was measured using automated microscopy (DMIPS cell analyser) in the presence or absence of three known modifiers of DNA repair processes over the X-ray dose range 0-1 Gy. RESULTS: 3-Aminobenzamide (5 microM), a potent inhibitor of poly(ADP ribose)-polymerase, inhibited the development of increased radioresistance as indicated by a statistically significant reduction in an RBE from 3.537 (+/-SEM (0.139)) to 2.168 (+/-0.191) at an X-ray dose of 1 Gy, implying an involvement of DNA repair pathways that require poly(ADP ribose)-polymerase in the IRR response. In contrast, novobiocin (350 microM), an inhibitor of topoisomerase II, did not inhibit the development of increased radioresistance (RBE 3.650 (+/-0.192) to 3.322+/-(0.156)) but eliminated low-dose hypersensitivity as measured by an increase in RBE from 2.508 (+/-0.536) to 1.135 (+/-0.057) at 0.04 Gy. Ara-A (120 microM), an inhibitor of DNA polymerase, sensitized cells at all doses. CONCLUSION: These data support the hypothesis that DNA repair processes are likely to be involved in the development of increased radioresistance and provide further evidence against a sensitive subpopulation explanation for the biphasic low-dose survival response.

Animals↗
PubMed2000

Cell-cell contact increases radioresistance in head and neck carcinoma cell lines.

PURPOSE: To investigate the in vitro radiosensitivity of microcolonies from head and neck cancers of varying degrees of differentiation. MATERIALS AND METHODS: The location of individual squamous-cell carcinoma (SCC) cells in tissue culture flasks was recorded using computerized microscopy. This allowed the positions of the cells to be continually revisited, which enabled manual sizing of individual microcolonies, both at the time of irradiation (0-5 Gy) and following a post-irradiation incubation period during which the microcolonies were assessed for clonogenic survival. RESULTS: When irradiated as microcolonies with 0-3 Gy, malignant cells had plating efficiencies (PE; applying multiplicity corrections) that were lower than expected when compared with the PE of cells irradiated individually. However, at between 2 Gy and 3 Gy the PE values became similar and at higher doses the 'corrected PE' of microcolonies became higher than that of individually irradiated cells. CONCLUSION: These data show that cell cell interactions can occur in malignant cells in a manner similar to that demonstrated previously in non-malignant cells. Cells within microcolonies demonstrate increased radioresistance compared with cells irradiated singly.

Carcinoma, Squamous Cell↗
PubMed1999

Effect of serum starvation on expression and phosphorylation of PKC-alpha and p53 in V79 cells: implications for cell death.

The effect of serum starvation on the expression and phosphorylation of PKC-alpha and p53 in Chinese hamster V79 cells was investigated. Serum starvation led to growth arrest, rounding up of cells and the appearance of new PKC-alpha and p53 bands on Western blots. Prolonged incubation (> or = 48 hr) in serum-deprived medium led to cell detachment and death. Moving cells to fresh medium containing 10% serum before, but not after, cell detachment reversed the changes observed in PKC-alpha and p53, and also prevented later cell detachment. Radiolabelling studies showed that the higher-molecular-weight PKC-alpha and p53 bands result from increased phosphorylation, while a lower-molecular-weight PKC-alpha band reflects newly synthesized protein. Immunocomplex kinase assays have shown that the increased phosphorylation of PKC-alpha is associated with its increased activity. To study the relationship between PKC-alpha, p53 and cell death, cells were treated either with TPA, to down-regulate PKC or with staurosporine, to inhibit PKC activity. Staurosporine, a potent PKC inhibitor and inducer of programmed cell death, caused the appearance of new PKC-alpha and p53 bands similar to those induced by serum starvation. If serum starvation was preceded by prolonged (48 hr) TPA treatment to down-regulate PKC-alpha, cell detachment and death did not take place within the same time frame. Intracellular fractionation of cells demonstrated that increased expression of PKC-alpha and the appearance of the associated higher and lower molecular-weight bands occurred in the nucleus. These data highlight the association of PKC-alpha and p53 with cellular events leading to cell death.

Animals↗
PubMed1999

The response of human glioma cell lines to low-dose radiation exposure.

PURPOSE: To examine the low-dose radiation response of a series of radioresistant human glioma cell lines and determine if low-dose hypersensitivity is a characteristic of these cells. MATERIALS AND METHODS: The clonogenic survival of six radioresistant human glioma cell lines was measured following exposure to graded, single, very low doses of X-rays in vitro. High resolution was achieved using either a Dynamic Microscopic Image Processing Scanner (DMIPS) or a cell sorter (CS). RESULTS: In five of the six cell lines tested, low-dose hypersensitivity (HRS) was demonstrated although in the sixth, a grade III astrocytoma line, it was not. These results are consistent with previous data indicating that low-dose hypersensitivity is more marked in more radioresistant cell lines although the difference between the glioblastoma cell lines with differing SF2 is not marked. CONCLUSION: Low-dose hypersensitivity is common in radioresistant glioma cell lines. This may have implications for the treatment of these tumours if further studies confirm that HRS translates to increased effectiveness per gray in vivo when very low doses per fraction are used.

Astrocytoma↗
PubMed1999

Low dose hypersensitivity in the T98G human glioblastoma cell line.

PURPOSE: To examine the low dose-response of a human radioresistant glioblastoma cell line (T98G) using two different methods to measure surviving fraction and to define the influence of cell cycle phase on this response. MATERIALS AND METHODS: The survival of cells following exposure to single very low doses of X-rays in vitro was measured using either the Dynamic Microscopic Image Processing Scanner (DMIPS) or a Cell Sorter (CS). The DMIPS was also used to measure the low dose survival response of T98G cells following manipulation of their progression through the cell cycle. RESULTS: With both methods, T98G demonstrated marked low dose hyper-radiosensitivity (HRS) and the two methods produced very similar data in the low dose region of the survival curve. However, the CS protocol produced less variable results and was the more efficient method of generating low dose data. HRS was also demonstrated when these cells were irradiated while held in reversible arrest in the G1 phase of the cell cycle, but the effect was less marked than in the asynchronous population. CONCLUSIONS: T98G glioblastoma cells demonstrate marked HRS, which is a characteristic of the whole population rather than being due to the influence of a small subpopulation of hyper-radiosensitive cells within a particular phase of the cell cycle.

Cell Cycle↗
PubMed1999

The effect of microcolony size, at time of irradiation, on colony forming ability.

PURPOSE: To investigate the effect of irradiating microcolonies of various sizes on their subsequent ability to form colonies. MATERIALS AND METHODS: The location of individual HPV-G cells in tissue culture flasks was recorded using computerized microscopy. This allowed the cells' positions to be continually revisited, which enabled manual sizing of individual microcolonies both at the time of irradiation (between 0-5 Gy) and following an incubation period during which the microcolonies were assessed for cologenic survival. RESULTS: The experiments indicate that when irradiated as microcolonies, cells have plating efficiencies (PE) that are lower than expected when compared with the prediction from the PE values of cells irradiated individually. CONCLUSION: These data support the conclusions of Mothersill and Seymour (1997b) which challenge the concept of independent survival of certain types of cells following irradiation. The results indicate that interactions between cells can occur even in small aggregates.

Cell Aggregation↗
PubMed1999

A novel human stress response-related gene with a potential role in induced radioresistance.

We have isolated a novel gene, DIR1, from L132 cells that is transiently repressed after exposure to low radiation doses and has a potential role in induced radioresistance. Molecular and cellular characterization of this gene reveals that it is unique but has similarities to a family of heat-shock-related proteins known as immunophilins. These have been implicated in various cellular functions including general stress responses and control of the cell cycle. Antisense strategies have demonstrated that the DIR1 gene also appears to have some involvement in the control of the cell cycle. Furthermore, there appears be a potential role for this gene product in the phenomenon of induced radioresistance through a mechanism that increases the rate of DNA repair in cells exposed to X rays and subsequently increases the cells' resistance to radiation. This is the first description of an immunophilin-like gene that has a possible role in adaptive/inducible responses to X rays in mammalian cells.

Amino Acid Sequence↗
PubMed1998

Hypoxia facilitates tumour cell detachment by reducing expression of surface adhesion molecules and adhesion to extracellular matrices without loss of cell viability.

The effects of acute hypoxia on integrin expression and adhesion to extracellular matrix proteins were investigated in two human melanoma cell lines, HMB-2 and DX3, and a human adenocarcinoma cell line, HT29. Exposure to hypoxia caused a significant down-regulation of cell surface integrins and an associated decrease in cell adhesion. Loss of cell adhesion and integrin expression were transient and levels returned to normal within 24 h of reoxygenation. Other cell adhesion molecules, such as CD44 and N-CAM, were also down-regulated after exposure of cells to hypoxia. Acute exposure to hypoxia of cells at confluence caused rapid cell detachment. Cell detachment preceded loss of viability. Detached HMB-2 and DX3 cells completely recovered upon reoxygenation, and floating cells re-attached and continued to grow irrespective of whether they were left in the original glass dishes or transferred to new culture vessels, while detached HT29 cells partly recovered upon reoxygenation. Cell detachment after decreased adhesion appears to be a stress response, which may be a factor enabling malignant cells to escape hypoxia in vivo, with the potential to form new foci of tumour growth.

Cell Adhesion↗
PubMed1997

The Klaas Breur Lecture. Radiation, hypoxia and genetic stimulation: implications for future therapies.

The cellular stress response, whereby very low doses of cytotoxic agents induce resistance to much higher doses, is an evolutionary defence mechanism and is stimulated following challenges by numerous chemical, biological and physical agents including particularly radiation, drugs, heat and hypoxia. There is much homology in the effects of these agents which are manifest through the up-regulation of various genetic pathways. Low-dose radiation stress influences processes involved in cell-cycle control, signal transduction pathways, radiation sensitivity, changes in cell adhesion and cell growth. There is also homology between radiation and other cellular stress agents, particularly hypoxia. Whereas traditionally, hypoxia was regarded mainly as an agent conferring resistance to radiation, there is now much evidence illustrating the cytokine-like properties of hypoxia as well as radiation. Stress phenomena are likely to be important in risks arising from low doses of radiation. Conversely, exploitation of the stress response in settings appropriate to therapy can be particularly beneficial not only in regard to radiation alone but in combinations of radiation and drugs. Similarly, tissue hypoxia can be exploited in novel ways of enhancing therapeutic efficacy. Bioreductive drugs, which are cytotoxically activated in hypoxic regions of tissue, can be rendered even more effective by hypoxia-induced increased expression of enzyme reductases. Nitric oxide pathways are influenced by hypoxia thereby offering possibilities for novel vascular based therapies. Other approaches are discussed.

Cell Hypoxia↗