PubMed Health⌕ Search

Biomedical subjects

C R Yang

Publications and source records attributed to C R Yang.

At least 37 records · Page 2Linked to original sources
PubMed2001

The role of lymphadenectomy in the radical nephrectomy for renal cell carcinoma.

BACKGROUND: The value of lymphadenectomy in the management of renal cell carcinoma (RCC) remains controversial. Most would agree that lymphadenectomy (LD) provides accurate pathologic staging for prognostic data. The question remains whether there is any significant therapeutic benefit from extensive lymph node dissection with radical nephrectomy. The aim of this study was to compare outcomes in sampled lymphadenectomy (SLD) and extensive lymphadenectomy (ELD) with radical nephrectomy for RCC. METHODS: One hundred and thirty-seven patients with RCC were enrolled in this study from Oct. 1982 to Dec. 1996. Eighty-one patients received radical nephrectomy with SLD (stage I: 43, II: 16, III: 22). Fifty-six patients received radical nephrectomy with ELD (stage I: 30, II: 11, III: 15). RESULTS: The mean number of lymph nodes removed by SLD was 4 (ranged from 1 to 8). The mean number of lymph nodes removed by ELD was 16.1 (ranged from 9 to 32). Overall 5-year survivals for SLD in stages I, II and III were 98%, 80% and 38%, respectively. Overall 5-year survivals for ELD in stages I, II and III were 92%, 84% and 40%, respectively. CONCLUSIONS: There is no significant therapeutic benefit from ELD in patients with RCC receiving radical nephrectomy.

Adult↗
PubMed2001

[Cochinchinenin--a new chalcone dimer from the Chinese dragon blood].

AIM: To study the active constituents of Dracaena cochinchinensis (Lour.) S.C. Chen. in the commercial dragon blood. METHODS: Various column chromatographies with Sephadex L-20 gel, MCI gel and silica gel were employed for the isolation and purification. The structures of compounds were elucidated by spectral analysis. RESULTS: Nine chalcones were isolated from the commercial dragon's blood which was made of D. cochinchinensis (Lour.) S.C. Chen.. By means of spectral data, they were identified as 1-[5-(2,4,4'-trihydroxydihydrochalconyl)]-1- (p-hydroxyphenyl)-3-(2-methoxy-4-hydroxy-phenyl)-propane (1), 2'-methoxysocotrin-5'-ol (2), socotrin-4'-ol (3), 2-methoxy-4, 4'-dihydroxydihydrochalcone (4), 2, 4, 4'-trihydroxy-dihydrochalcone (5), 2, 4, 4'-trihydroxy-6-methoxydihydrochalcone (6), 2', 4', 4-trihydroxychalcone (7), 2-methoxy-4, 4'-dihydroxychalcone (8) and 2'-methoxy-4', 4-dihydroxychalcone (9). CONCLUSION: Compound 1 is a new chalcone dimer and named as cochinchinenin. Compounds 2-9 were isolated from D. cochinchinensis (Lour.) S.C. Chen. for the first time.

Chalcone↗
PubMed2000

Nuclear clusterin/XIP8, an x-ray-induced Ku70-binding protein that signals cell death.

Clusterin [CLU, a.k.a. TRPM-2, SGP-2, or ionizing radiation (IR)-induced protein-8 (XIP8)] was implicated in apoptosis, tissue injury, and aging. Its function remains elusive. We reisolated CLU/XIP8 by yeast two-hybrid analyses using as bait the DNA double-strand break repair protein Ku70. We show that a delayed (2-3 days), low-dose (0.02-10 Gy) IR-inducible nuclear CLU/XIP8 protein coimmunoprecipitated and colocalized (by confocal microscopy) in vivo with Ku70/Ku80, a DNA damage sensor and key double-strand break repair protein, in human MCF-7:WS8 breast cancer cells. Overexpression of nuclear CLU/XIP8 or its minimal Ku70 binding domain (120 aa of CLU/XIP8 C terminus) in nonirradiated MCF-7:WS8 cells dramatically reduced cell growth and colony-forming ability concomitant with increased G(1) cell cycle checkpoint arrest and increased cell death. Enhanced expression and accumulation of nuclear CLU/XIP8-Ku70/Ku80 complexes appears to be an important cell death signal after IR exposure.

Adenocarcinoma↗
PubMed2000

Presynaptic GABA(B) receptors modulate organum vasculosum lamina terminalis-evoked postsynaptic currents in rat hypothalamic supraoptic neurons.

Whole-cell patch-clamp recordings obtained from 36 hypothalamic supraoptic nucleus neurons in explant preparations evaluated a role for GABA(B) receptors in modulating postsynaptic inhibitory and excitatory currents evoked by electrical stimulation in the organum vasculosum of the lamina terminalis. At a holding current of -65 mV, application of baclofen (1-10 microM) induced a dose-dependent reduction in the amplitude of pharmacologically isolated inhibitory and excitatory postsynaptic currents, converted paired-pulse depression in inhibitory postsynaptic currents to paired-pulse facilitation, and enhanced paired-pulse ratios for excitatory postsynaptic currents. In media containing 2-hydroxysaclofen (200-400 microM), baclofen-associated events were blocked and paired-pulse depression in evoked inhibitory postsynaptic currents was abolished. In addition, a progressive increase in the amplitude of inhibitory postsynaptic currents implied that GABA was endogenously active at presynaptic GABA(B) receptors. In contrast, no paired-pulse depression was observed for inhibitory postsynaptic currents evoked in six non-magnocellular neurons. Neither baclofen nor 2-hydroxysaclofen altered holding currents or input resistances in supraoptic neurons, or altered the kinetics of the evoked responses.These observations imply that the terminals of both inhibitory (GABAergic) and excitatory (glutamatergic) afferents to supraoptic nucleus neurons from organum vasculosum lamina terminalis neurons are subject to modulation by presynaptic GABA(B) receptors, and that this modulation is preferentially directed to the inhibitory inputs.

Animals↗
PubMed2000

Novel norsesquiterpenoids from the roots of Phyllanthus emblica.

Three ester glycosides, named phyllaemblicins A (3), B (4), and C (5), and a methyl ester (2), of a highly oxygenated norbisabolane, phyllaemblic acid (1), were isolated from the roots of Phyllanthus emblica, along with 15 tannins and related compounds. The structures of 2-5 were established by spectral and chemical methods.

China↗
PubMed2000

Coordinate modulation of Sp1, NF-kappa B, and p53 in confluent human malignant melanoma cells after ionizing radiation.

Regulation of transcriptional responses in growth-arrested human cells under conditions that promote potentially lethal damage repair after ionizing radiation (IR) is poorly understood. Sp1/retinoblastoma control protein (RCP) DNA binding increased within 30 min and peaked at 2-4 h after IR (450-600 cGy) in confluent radioresistant human malignant melanoma (U1-Mel) cells. Increased phosphorylation of Sp1 directly corresponded to Sp1/RCP binding and immediate-early gene induction, whereas pRb remained hypophosphorylated. Transfection of U1-Mel cells with the human papillomavirus E7 gene abrogated Sp1/RCP induction and G(0)/G(1) cell cycle checkpoint arrest responses, increased apoptosis and radiosensitivity, and augmented genetic instability (i.e., increased polyploidy cells) after IR. Increased NF-kappaB DNA binding in U1-Mel cells after IR treatment lasted much longer (i.e., >20 h). U1-Mel cells overexpressing dominant-negative IkappaBalpha S32/36A mutant protein were significantly more resistant to IR exposure and retained both G(2)/M and G(0)/G(1) cell cycle checkpoint responses without significant genetic instability (i.e., polyploid cell populations were not observed). Nuclear p53 protein levels and DNA binding activity increased only after high doses of IR (>1200 cGy). Disruption of p53 responses in U1-Mel cells by E6 transfection also abrogated G(0)/G(1) cell cycle checkpoint arrest responses and increased polyploidy after IR, but did not alter radiosensitivity. These data suggest that abrogation of individual components of this coordinate IR-activated transcription factor response may lead to divergent alterations in cell cycle checkpoints, genomic instability, apoptosis, and survival. Such coordinate transcription factor activation in human cancer cells is reminiscent of prokaryotic SOS responses, and further elucidation of these events should shed light on the initial molecular events in the chromosome instability phenotype.-Yang, C.-R., Wilson-Van Patten, C., Planchon, S. M., Wuerzberger-Davis, S. M., Davis, T. W., Cuthill, C., Miyamoto, S., Boothman, D. A. Coordinate modulation of Sp1, NF-kappa B, and p53 in confluent human malignant melanoma cells after ionizing radiation.

Cell Cycle↗
PubMed2000

Dopamine D1/D5 receptor activation modulates a persistent sodium current in rat prefrontal cortical neurons in vitro.

The effects of dopamine (DA) on a persistent Na(+) current (I(NaP)) in layer V-VI prefrontal cortical (PFC) pyramidal cells were studied using whole cell voltage-clamp recordings in rat PFC slices. After blocking K(+) and Ca (2+) currents, a tetrodotoxin-sensitive I(NaP) was activated by slow depolarizing voltage ramps or voltage steps. DA modulated the I(NaP) in a voltage-dependent manner: increased amplitude of I(NaP) at potentials more negative than -40 mV, but decreased at more positive potentials. DA also slowed the inactivation process of I(NaP). The D1/D5 dopamine receptor agonists SKF 38393, SKF 81297, and dihydrexidine (3-10 microM), but not the dopamine D2/D3 receptor agonist qiunpirole (1-20 microM), mimicked the effects of DA on I(NaP). Modulation of I(NaP) by D1/D5 agonists was blocked by the D1/D5 antagonist SCH23390. Bath application of specific protein kinase C inhibitor, chelerhythrine, or inclusion of the specific protein kinase C inhibiting peptide([19-36]) in the recording pipette, but not protein kinase A inhibiting peptide([5-24]), blocked the effect of D1/D5 agonists on I(NaP). In current-clamp recordings, D1/D5 receptors activation enhanced the excitability of cortical pyramidal cells. Application of the D1/D5 agonist SKF 81297 induced a long-lasting decrease in the first spike latency in response to depolarizing current ramp. This was associated with a shift in the start of nonlinearity in the slope resistance to more negative membrane potentials. We proposed that this effect is due to a D1/D5 agonist-induced leftward shift in the activation of I(NaP). This enables DA to facilitate the firing of PFC neurons in response to depolarizing inputs.

2,3,4,5-Tetrahydro-7,8-dihydroxy-1-phenyl-1H-3-ben↗
PubMed2000

Ultrasonographic study of carotid artery structural changes with natural longevity.

The changes in carotid artery structure during the natural longevity of the Uighurs people were compared with other ethnic groups. The 419 subjects were divided into (1) longevity and (2) older groups as follows: Uighurs longevity group in Hotan (ULH); and Uighurs, Hans and Kazaks older groups in Hotan or Balikun (UOH, HOH, HOB and KOB). The wall thickness of the common carotid artery (CCA) was measured by ultrasonography. Risk factors were evaluated by measuring total cholesterol (T-C), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), triglyceride (TG) and fasting glucose, and by 24-h ambulatory blood pressure monitoring. Wall thickness and blood pressure were greater in the Balikun groups than in the Hotan groups. The CCA wall was thicker in Uighurs and Kazaks than in Hans, in the ULH than in the UOH. There was no significant difference in blood pressure among the Hotan groups. LDL-C and glucose were higher and HDL-C was lower in the Hotan groups than in the Balikun groups. T-C, LDL-C, TG and glucose were higher in the HOH than in the ULH and UOH. Systolic blood pressure and LDL-C were higher and TG was lower in the KOB than in the HOB. These results suggest that an increased CCA wall thickness can be attributed to age in Hotan, and to a higher blood pressure level in Balikun, and that the higher blood pressure affected the Balikun groups more than the Hotan groups.

Aged↗
PubMed1999

Medial prefrontal cortical output neurons to the ventral tegmental area (VTA) and their responses to burst-patterned stimulation of the VTA: neuroanatomical and in vivo electrophysiological analyses.

During a delayed period in a delayed-response task, prefrontal cortical neurons show a change in neuronal firing rate that is dependent on a functional mesocortical dopamine input. This change in firing rate has been attributed to be part of the cellular processes underlying working memory. However, it is unclear what neural mechanisms activate mesocortical dopamine neurons to provide an optimal level of dopamine to modulate the firing of the medial prefrontal cortical (mPFC) neurons. This study examined the possibility of whether mPFC neurons that project to the ventral tegmental area (VTA) might activate the ascending mesocortical dopamine neurons. To determine the locations of the mPFC-->VTA neurons, cholera toxin subunit B was microinjected into the VTA. Retrogradely labeled mPFC neurons mainly reside in the deep lamina V and VI. In vivo single unit recording in urethane-anesthetized rats were also used to determine the responses of some of these neurons to burst-patterned stimulation of the VTA. Single-pulse stimulation (1 Hz) of the VTA antidromically activated burst firing mPFC-->VTA neurons. In response to burst-patterned stimulation of the VTA, which mimicked burst firing of VTA dopamine neurons (4-10 pulses at 10-15 Hz cycled at 0.5-3 Hz), the temporal structure of spontaneous burst firing patterns of these neurons but not their mean firing rate were changed. However, the mean firing rate of the non-VTA projecting neurons (i.e., no antidromic response to VTA stimulations) was either increased or decreased by similar burst-patterned stimulation of the VTA. These data suggest that burst-patterned stimulation of the ascending VTA-->mPFC or putative mesocortical dopamine neurons might have released dopamine and/or other neuromodulators to modulate the temporal code, rather than the rate code, of mPFC-->VTA neurons. Medial PFC neurons that project elsewhere (e.g., nucleus accumbens or mediodorsal thalamus) may mediate the sustained firing rate changes during, e.g., short-term working memory.

Animals↗
PubMed1999

Isolation of Ku70-binding proteins (KUBs).

DNA-dependent protein kinase (DNA-PK) plays a critical role in resealing DNA double-stand breaks by non-homologous end joining. Aside from DNA-PK, XRCC4 and DNA ligase IV, other proteins which play a role(s) in this repair pathway remain unknown; DNA-PK contains a catalytic subunit (DNA-PKcs) and a DNA binding subunit (Ku70 and Ku80). We isolated Ku70-binding proteins (KUB1-KUB4) using yeast two-hybrid analyses. Sequence analyses revealed KUB1 to be apolipoprotein J (apoJ), also known as X-ray-inducible transcript 8 (XIP8), testosterone-repressed prostate message-2 (TRPM-2) and clusterin. KUB2 is Ku80. KUB3 and KUB4 are unknown, >10 kb trans-cripts. Interactions of apoJ/XIP8 or KUB3 with Ku70 were confirmed by co-immunoprecipitation analyses in MCF-7:WS8 breast cancer or IMR-90 normal lung fibroblast cells, respectively. The interaction of apoJ/XIP8 with Ku70 was confirmed by far-western analyses. Stable over-expression of full-length apoJ/XIP8 in MCF-7:WS8 caused decreased Ku70/Ku80 DNA end binding that was restored by apoJ/XIP8 monoclonal antibodies. The role of apoJ/XIP8 in ionizing radiation resistance/sensitivity is under investigation.

Animals↗
PubMed1999

Protein kinase C and intracellular calcium are involved in follicle-stimulating hormone-mediated meiotic resumption of cumulus cell-enclosed porcine oocytes in hypoxanthine-supplemented medium.

The present experiments were conducted to examine the hypothesis that follicle-stimulating hormone (FSH) can stimulate the hydrolysis of phosphoinositide, generating the intracellular second messengers to activate protein kinase C and mobilizing intracellular calcium, thus inducing oocyte meiotic resumption. Pig cumulus cell-enclosed oocytes (CEO) were cultured for 24 hr in 4 mM hypoxanthine (HX)-supplemented medium and treated with different agents in the following designs: (1) CEO were treated with neomycin (an inhibitor of phosphoinositide hydrolysis) in the presence of FSH or only treated with 7,12-dimethylbenzin(a) anthracene (DMBA, a tumor promoter which can cause phosphorylation of phospholipase C (PLC), formation of inositol triphophate, and mobilization of intracellular calcium) to mimic the direct activation of PLC; (2) CEO were challenged by FSH, together with sphingosine or staurosporine (two kinds of PKC inhibitors); or treated with phorbol myristate acetate (PMA, an activator of PKC) separately; (3) CEO were primed with BAPTA/AM (an intracellular calcium chelator) or BAPTA/AM +FSH for 60 min, and then transferred into a new culture medium supplemented with FSH but without BAPTA/AM; total culture time was 24 hr. At the end of the culture, the incidence of germinal vesicle breakdown (GVBD) was calculated. The results showed that: (1) FSH (100 U/liter) could stimulate pig CEO to override the arrest of HX and resume meiosis; DMBA (10(-8)-10(-5) M) itself also had such a kind of effect; whereas neomycin, at the level of 10-20 mM, could dramatically inhibit the stimulatory effect of FSH. (2) Staurosporine (10(-9)-10(-6) M) or sphingosine (10(-8)-10(-5) M) could also inhibit the effect of FSH in a dose-dependent manner on stimulating CEO to resume meiosis. However, PMA (10(-8)-10(-5) M) alone had a dual effect on the meiotic resumption of pig CEO. PMA, at the level of 10(-8)-10(-6) M, could stimulate CEO to resume meiosis, and at high concentration of 10(-5) M , it could even enhance the inhibitory effect of HX. (3) Priming CEO with BAPTA/AM only or BAPTA/AM +FSH for 60 min could significantly inhibit the effect of FSH in a dose-dependent manner. These results indicate that in the process of ligand-mediated meiotic resumption of pig CEO, FSH can stimulate the hydrolysis of phosphoinositide leading to the activation of PKC and mobilization of intracellular calcium; and suggest that multiple signaling pathways and signal interaction are involved in this process.

Animals↗
PubMed1999

Hypothalamic A14 and A15 catecholamine cells provide the dopaminergic innervation to the supraoptic nucleus in rat: a combined retrograde tracer and immunohistochemical study.

This study investigated the origin of a dopaminergic innervation of the hypothalamic supraoptic nucleus. In pentobarbital-anaesthetized male Long-Evans rats, a transpharyngeal approach was used to inject a retrograde tracer, rhodamine latex microspheres, into the supraoptic nucleus. After 13-26 h survival under anaesthesia, animals were perfused transcardially, the brain sectioned and processed for tyrosine hydroxylase immunofluorescence, a marker for hypothalamic dopaminergic neurons. In six cases with injections restricted to the supraoptic nucleus, rhodamine-labelled microspheres were observed in a population of tyrosine hydroxylase-positive neurons located in the A15 cells below the anterior commissure (A15 dorsal) and above the optic chiasm (A15 ventral), and the dorsal and lateral periventricular A14 cell group. Occasional double-labelled cells were seen in the medial and lateral hypothalamus and bed nucleus of the stria terminalis, but rarely in other known dopaminergic cell groups, notably the ventral tegmental area (A10), zona incerta (A13) and substantia nigra. In support of a role for dopamine in neurohypophysial regulation, these observations indicate that the major dopaminergic input to magnocellular neurons in the hypothalamic supraoptic nucleus is derived from a relatively sparse population of neurons located in the A14 and A15 cell groups.

Animals↗
PubMed1999

Developing a neuronal model for the pathophysiology of schizophrenia based on the nature of electrophysiological actions of dopamine in the prefrontal cortex.

This review covers some recent findings of the electrophysiological mechanisms through which mesocortical dopamine modulates prefrontal cortical neurons. Dopamine has been shown to modulate several ionic conductances located along the soma-dendritic axis of prefrontal cortical pyramidal neurons. These ionic currents include high-voltage-activated calcium currents and slowly inactivating Na+ and K+ currents. They contribute actively in processing functionally segregated inputs during synaptic integration. In addition, dopamine mainly depolarizes the fast-spiking subtype of local GABAergic interneurons that connect the pyramidal neurons. This latter action can indirectly control pyramidal cell excitability. These electrophysiological data indicate that the actions of dopamine are neither "excitatory" nor "inhibitory" in pyramidal prefrontal cortex neurons. Rather, the actions of dopamine are dependent on somadendritic loci, timing of the arrival of synaptic inputs, strength of synaptic inputs, as well as the membrane potential range at which the PFC neuron is operating at a given moment. Based on available electrophysiological findings, a neuronal model of the pathophysiology of schizophrenia is presented. This model proposes that episodic hypo- and hyperactivity of the PFC and the associated dysfunctional mesocortical dopamine system (and their interconnected brain regions) may coexist in the same schizophrenic patient in the course of the illness. We hypothesize that the dysfunctional mesocortical dopamine input to the PFC may lead to abnormal modulation of ionic channels distributed in the dendritic-somatic compartments of PFC pyramidal neurons that project to the ventral tegmental area and/or nucleus accumbens. In some schizophrenics, a reduction of mesocortical dopamine to below optimal levels and/or a loss of local GABAergic inputs may result in a dysfunctional integration of extrinsic associative inputs by Ca2+ channel activity in the distal dendrites of PFC pyramidal neurons. This may account for the patients' distractibility caused by their inability to focus only on relevant external inputs. In contrast, in acute stress or psychotic episodes, an associated abnormal elevation of mesocortical dopamine transmission may greatly influence distal dendritic Ca2+ channel-mediated signal-processing mechanisms. This can enhance possible reverberative activity between adjacent interconnected pyramidal neurons via the effects of dopamine on the slowly inactivating Na+, K+, and soma-dendritic Ca2+ currents. The effects of high levels of PFC dopamine in this case may contribute to behavioral perseveration and stereotypy so that the patients are unable to use new external cues to modify ongoing behaviors.

Animals↗
PubMed1999

Safety of distilled water as an irrigating fluid for transurethral resection of the prostate.

BACKGROUND: Irrigating fluid is necessary for transurethral resection of the prostate (TURP). Initially, distilled water was used as irrigating fluid and some advantages and complications were noted. Although other solutions were used to substitute for distilled water, we collected data for changes in electrolytes and hematocrit (Hct) during surgery to evaluate the safety of distilled water as irrigating fluid for TURP with suprapubic trocar cystostomy. METHODS: Fifty-three patients who underwent TURP using distilled water as irrigating fluid were investigated. TURP was performed with suprapubic trocar cystostomy routinely. Preoperative and postoperative blood samples were collected and analyzed for electrolytes and Hct to study any changes during surgery. RESULTS: The change in plasma sodium concentration was not statistically significant, but the decreases in blood Hct and plasma potassium concentration were statistically significant. There was no major operative complication and no clinical-evident related postoperative renal impairment. CONCLUSIONS: Distilled water is a relatively safe solution as an irrigating fluid for TURP under drainage with suprapubic trocar cystostomy, but attention must be paid to the inevitable blood loss during surgery.

Aged↗
PubMed1999

Long-term results with pediatric kidney transplants in adult recipients.

The shortage of donor kidneys available for transplantation has resulted in an expansion of the criteria used for donor selection. The use of cadaveric pediatric kidneys has been suggested as a means to overcome this shortage, but is debated because of technical complications and an increased incidence of functional allograft impairment. We examined the records of all adult patients receiving renal allograft transplants from January, 1983 through December 1994, to determine whether kidneys from pediatric donors can be used safely for transplantation. During the study period, 204 adult patients received kidney transplants. The patients were divided into three groups by donor age: group 1, 26 months to 7 years old (n = 12); group 2, 7 to 14 years old (n = 19); and group 3, adult donors (n = 173). Cyclosporine- and/or azathioprine-based immunosuppression regimens were used. The three groups did not differ from one anther with respect to recipient age, recipient sex, primary renal disease, human leukocyte antigen (HLA) mismatching, or the proportion of recipients with previous transplants. The graft survival rates at 1 and 3 years, respectively, were 75% and 75% in group 1,84% and 79% in group 2, and 85% and 72% in group 3 (p > 0.05). The patient survival rates at 1 and 3 years, respectively, were 75% and 75% in group 1,95% and 90% in group 2, and 90% and 83% in group 3 (p > 0.05). The frequency of fixed proteinuria (> or = 0.8 gm/day) in the first 3 years was higher in the two pediatric groups (50%) than in the adult group (28%) (p < 0.05). The frequency of surgical complications was higher with pediatric grafts (group 1 vs group 3: 67% vs 24%, p < 0.005). In particular, group 1 patients had much higher rates of lymphocele (50% vs 8%), renal graft artery stenosis (33% vs 11%), and hydronephrosis (33% vs 9%) than group 3. We conclude that pediatric donor kidneys can be used with reasonable safety, although surgical complications remain a major problem.

Adult↗
PubMed1998

Apoptosis, reproductive failure, and oxidative stress in Chinese hamster ovary cells with compromised genomic integrity.

Chromosomal instability and persistent reproductive cell death show a significant correlation after cells are exposed to ionizing radiation. To examine the possible role of apoptosis in persistent reproductive cell death, we analyzed subsets of chromosomally stable and unstable clones for relationships between chromosome stability, reproductive integrity, and apoptosis. All clones were generated from the GM10115 cell line and derived from single progenitor cells surviving 10 Gy of X-rays, and all measurements were made approximately 60-80 generations after irradiation. The incidence of apoptosis, as measured by both annexin V binding of phosphatidylserine residues and terminal deoxynucleotidyl transferase labeling of DNA strand breaks, was significantly higher in chromosomally unstable clones than it was in chromosomally stable clones (P < 0.05; ANOVA and Student's t test). Furthermore, statistical analyses of the biological end points of persistent reproductive cell death and apoptosis were consistent, showing R2 values of 0.78 and 0.76, respectively. These results suggest that persistent reproductive cell death can, in part, be explained by the predisposition of a fraction of the clonal population to undergo apoptosis or necrosis. Immunological blot analyses of protein levels and DNA bandshift assays confirmed the mutant status of p53 in the host cell line, implying an apoptotic pathway that is independent of p53. Induction of apoptosis by agents such as actinomycin D, etoposide, and staurosporine and induction of necrosis by sodium azide were accompanied by an increase in the level of intracellular peroxy radicals and lipid peroxidation products, two independent end points that are typically associated with oxidative stress. Similar findings were observed in several subclones showing persistent apoptosis. These results suggest that the elevated levels of free radical damage that we detected were derived from the fraction of cells dying by apoptotic or necrotic processes. Possible mechanisms whereby oxidative stress may contribute indirectly to the perpetuation of chromosomal instability are discussed.

Animals↗