Biomedical subjects

M Gao

Publications and source records attributed to M Gao.

At least 73 records · Page 4Linked to original sources

PubMed2000

Interaction between a poly(A)-specific ribonuclease and the 5' cap influences mRNA deadenylation rates in vitro.

We have used an in vitro system that reproduces in vivo aspects of mRNA turnover to elucidate mechanisms of deadenylation. DAN, the major enzyme responsible for poly(A) tail shortening in vitro, specifically interacts with the 5' cap structure of RNA substrates, and this interaction is greatly stimulated by a poly(A) tail. Several observations suggest that cap-DAN interactions are functionally important for the networking between regulated mRNA stability and translation. First, uncapped RNA substrates are inefficiently deadenylated. Second, a stem-loop structure in the 5' UTR dramatically reduces deadenylation by interfering with cap-DAN interactions. Third, the addition of cap binding protein eIF4E inhibits deadenylation in vitro. These data provide insights into the early steps of substrate recognition that target an mRNA for degradation.

Eukaryotic Initiation Factor-4E↗

PubMed2000

Starch-branching enzymes preferentially associated with A-type starch granules in wheat endosperm.

Two starch granule-bound proteins (SGP), SGP-140 and SGP-145, were preferentially associated with A-type starch granules (>10 microm) in developing and mature wheat (Triticum aestivum) kernels. Immunoblotting and N-terminal sequencing suggested that the two proteins were different variants of SBEIc, a 152-kD isoform of wheat starch-branching enzyme. Both SGP-140 and SGP-145 were localized to the endosperm starch granules but were not found in the endosperm soluble fraction or pericarp starch granules younger than 15 d post anthesis (DPA). Small-size starch granules (<10 microm) initiated before 15 DPA incorporated SGP-140 and SGP-145 throughout endosperm development and grew into full-size A-type starch granules (>10 microm). In contrast, small-size starch granules harvested after 15 DPA contained only low amounts of SGP-140 and SGP-145 and developed mainly into B-type starch granules (<10 microm). Polypeptides of similar mass and immunologically related to SGP-140 and/or SGP-145 were also preferentially incorporated into A-type starch granules of barley (Hordeum vulgare), rye (Secale cereale), and triticale (x Triticosecale Wittmack) endosperm, which like wheat endosperm have a bimodal starch granule size distribution.

1,4-alpha-Glucan Branching Enzyme↗

PubMed2000

Early discharge of patients with presumed opioid overdose: development of a clinical prediction rule.

OBJECTIVE: To develop a clinical prediction rule to identify patients who can be safely discharged one hour after the administration of naloxone for presumed opioid overdose. METHODS: Patients who received naloxone for known or presumed opioid overdose were formally evaluated one hour later for multiple potential predictor variables. Patients were classified into two groups: those with adverse events within 24 hours and those without. Using classification and regression tree methodology, a decision rule was developed to predict safe discharge. RESULTS: Clinical findings from 573 patients allowed us to develop a clinical prediction rule with a sensitivity of 99% (95% CI = 96% to 100%) and a specificity of 40% (95% CI = 36% to 45%). Patients with presumed opioid overdose can be safely discharged one hour after naloxone administration if they: 1) can mobilize as usual; 2) have oxygen saturation on room air of >92%; 3) have a respiratory rate >10 breaths/min and <20 breaths/min; 4) have a temperature of >35.0 degrees C and <37.5 degrees C; 5) have a heart rate >50 beats/min and <100 beats/min; and 6) have a Glasgow Coma Scale score of 15. CONCLUSIONS: This prediction rule for safe early discharge of patients with presumed opioid overdose performs well in this derivation set but requires validation followed by confirmation of safe implementation.

Adult↗

PubMed2000

Evidence for controlled incorporation of herpes simplex virus type 1 UL26 protease into capsids.

Herpes simplex virus type 1 (HSV-1) capsids are initially assembled with an internal protein scaffold. The scaffold proteins, encoded by overlapping in-frame UL26 and UL26.5 transcripts, are essential for formation and efficient maturation of capsids. UL26 encodes an N-terminal protease domain, and its C-terminal oligomerization and capsid protein-binding domains are identical to those of UL26.5. The UL26 protease cleaves itself, releasing minor scaffold proteins VP24 and VP21, and the more abundant UL26.5 protein, releasing the major scaffold protein VP22a. Unlike VP21 and VP22a, which are removed from capsids upon DNA packaging, we demonstrate that VP24 (containing the protease domain) is quantitatively retained. To investigate factors controlling UL26 capsid incorporation and retention, we used a mutant virus that fails to express UL26.5 (DeltaICP35 virus). Purified DeltaICP35 B capsids showed altered sucrose gradient sedimentation and lacked the dense scaffold core seen in micrographs of wild-type B capsids but contained capsid shell proteins in wild-type amounts. Despite C-terminal sequence identity between UL26 and UL26.5, DeltaICP35 capsids lacking UL26.5 products did not contain compensatory high levels of UL26 proteins. Therefore, HSV capsids can be maintained and/or assembled on a minimal scaffold containing only wild-type levels of UL26 proteins. In contrast to UL26.5, increased expression of UL26 did not compensate for the DeltaICP35 growth defect. While indirect, these findings are consistent with the view that UL26 products are restricted from occupying abundant UL26.5 binding sites within the capsid and that this restriction is not controlled by the level of UL26 protein expression. Additionally, DeltaICP35 capsids contained an altered complement of DNA cleavage and packaging proteins, suggesting a previously unrecognized role for the scaffold in this process.

Blotting, Western↗

PubMed2000

Androgen-induced growth inhibition of androgen receptor expressing androgen-independent prostate cancer cells is mediated by increased levels of neutral endopeptidase.

Androgen-mediated growth repression of androgen-independent prostate cancer (AIPC) cells has been reported in androgen-independent PC-3 cells overexpressing the androgen receptor, and in androgen-independent derivatives of LNCaP cells that develop following prolonged culture in androgen-free media. Using two models of AIPC, PC3/AR cells and LNCaP-OM1 cells, a subclone of LNCaP cells derived by prolonged culturing in charcoal-stripped media, we investigated whether expression of neutral endopeptidase 24.11 (NEP), a cell-surface peptidase that cleaves and inactivates neuropeptides implicated in the growth of AIPC, is induced by androgen, and whether NEP contributes to the observed androgen-mediated growth repression. These cell lines each express high levels of androgen receptor. Culturing in dihyrotestosterone (DHT) resulted in a 30-56% (PC3) and 35-43% (LNCaP-OM1) decrease in cell number over 7 days concomitant with a significant increase in NEP enzyme specific activity. Northern analysis detected an increase in NEP transcripts following DHT treatment in PC3/AR cells. The addition of the NEP enzyme inhibitor phosphoramidon to PC3 and LNCaP-OM1 or the NEP competitive inhibitor CGS 24592 to LNCaP-OM1 blocked the increase in NEP enzyme activity and reversed the DHT-induced growth inhibition. Neither phosphoramidon or CGS 24592 alone inhibited cell growth. Furthermore, the reversal of growth inhibition in LNCaPOM1 cells was dose dependent on the concentration of CGS 24592. These data indicate that androgen-induced growth repression of AIPC cells PC3 and LNCaP-OM1 results in part from androgen-induced expression of NEP in these cells.

Androgens↗

PubMed2000

Telelearning in a partnership between a university faculty and a regional health authority: benefits, challenges and strategies.

The University of Calgary's Faculty of Medicine and the Calgary Regional Health Authority understand that telehealth is an evolving field requiring both academic enquiry and operational readiness. Both parties are committed to quality educational programmes--the Faculty through its commitment to excellence and the Authority with its charge to maintain and enhance such programmes. There are shared applications, multi-learner user groups, shared strategies to overcome distances and shared infrastructure--technologies, communication pathways and resources. Having embarked on a joint telelearning venture, we have learned a number of lessons. Central to progress has been an appreciation and respect for unique mandates, a spirit of trust and flexibility, an agreement on a set of principles, ongoing communication between and participation from the users and, at times, redirection. Questions being answered include the following. How well is this collaborative model working? How functional is it at this time of health reform and restructuring? Can one meet complementary telelearning goals within a faculty-health authority relationship? These all have implications for future success.

Canada↗

PubMed2000

No correlation of polymorphism of angiotensin-converting enzyme genes with left ventricular hypertrophy in essential hypertension.

To investigate the correlation of polymorphism of angiotensin-converting enzyme (ACE) genes with left ventricular hypertrophy in essential hypertension, 151 patients with essential hypertension were studied. ACE genotypes were determined by PCR technology and diastolic left ventricular diameter (DLVd), systolic left ventricular diameter (SLVd), interseptal ventricular thickness (IVS), and left ventricular posterior wall thickness (LVPW) were scanned by echocardiography. Left ventricular mass (LVM) and the left ventricular mass index (LVMI) were calculated from echocardiographic findings. Results revealed that DLVd, SLVd, IVS, LVPW, LVM, and LVMI of the DD genotype group were 49.9 +/- 5.6 mm, 30.5 +/- 6.5 mm, 11.2 +/- 1.6 mm, 11.7 +/- 1.5 mm, 259.5 +/- 62.1 g, 92.7 +/- 23.5 g/m2, respectively. DLVd, SLVd, IVS, LVPW, LVM, and LVMI of the ID genotype group were 8.9 +/- 5.3 mm, 31.5 +/- 5.2 mm, 11.4 +/- 1.7 mm, 11.9 +/- 1.6 mm, 261.3 +/- 70.3 g, and 94.9 +/- 25.8 g/m2, respectively, and DLVd, SLVd, IVS, LVPW, LVM, and LVMI of the II genotype group are 48.9 +/- 5.5 mm, 31.8 +/- 6.5 mm, 11.1 +/- 1.9 mm, 11.5 +/- 1.8 mm, 250.8 +/- 82.5 g and 90.8 +/- 30.1 g/m2 respectively. There was no significant difference between the ID, DD and II genotype groups as regards DLVd, SLVd, IVS, LVPW, LVM, and LVMI (p > 0.05). These findings indicate that there is no association between the ACE gene and left ventricular hypertrophy in essential hypertension occurring in the Chinese population.

Aged↗

PubMed2000

Comparative studies of diet-related factors and blood pressure among Chinese and Japanese: results from the China-Japan Cooperative Research of the WHO-CARDIAC Study. Cardiovascular Disease and Alimentary Comparison.

We aimed to compare the differences in diet-related factors and their associations with blood pressure (BP) between Chinese and Japanese. A total of 1,151 Chinese (M/F: 551/600) and 1,681 Japanese (782/899), aged 48-56 years, were studied using a multi-center cross-sectional study design. This work was a constituent part of the World Health Organization (WHO) Cardiovascular Disease and Alimentary Comparison (CARDIAC) Study. Measurements included in the present report were BP, body mass index (BMI), serum total cholesterol (TC), 24-h urinary sodium, potassium, calcium, magnesium, creatinine, 3-Methylhistidine (3MH, a marker of animal protein intake) and taurine (a marker of seafood intake) excretion levels. Results were as follows: (a) Japanese men had a significantly higher prevalence of hypertension than the Chinese (34.4% vs. 20.5%, p<0.01). After adjustment for age, Japanese men had a significantly higher mean systolic and diastolic BP (SBP, DBP), and Japanese women had a significantly higher DBP than the Chinese subjects overall (p<0.01, respectively). (b) Japanese had significantly higher mean BMI, TC and sodium excretion, and lower mean magnesium excretion than Chinese (p<0.01). (c) In the Japanese sample, multiple linear regression analyses (using a stepwise procedure) showed that SBP had a significant positive association with BMI and sodium excretion, and a significant negative association with magnesium excretion, while DBP had a significant positive association with BMI and a significant negative association with the 3MH to creatinine ratio (3MH/Cre). In the Chinese sample, both SBP and DBP showed a significant positive association with BMI and sodium, and a significant negative association with 3MH/Cre. In conclusion, Japanese had significantly higher mean BP than Chinese. The differences in BP may have been partly attributable to differences in various diet-related factors, particularly in BMI, sodium, magnesium-rich foods and animal protein intake, between the two populations.

Blood Pressure↗

PubMed2000

Induction of growth factors in rat cardiac tissue by X irradiation.

To investigate the relationship between angiogenic growth factors and endothelial enzyme activity in capillaries after injury of rat cardiomyocytes caused by X irradiation, 7-week-old male Wistar rats were anesthetized with pentobarbitone and their hearts irradiated (X rays, 20 Gy) through a hole in the lead casing in which they were enclosed. The hearts were excised at 1 h, 1 week and 3 weeks after irradiation. Left ventricular cross sections were stained for capillary enzymes by double staining for two endothelial enzymes, alkaline phosphatase (AP) and dipeptidylpeptidase IV (DPP), immunohistochemically stained for basic fibroblast growth factor (Fgf, also known as bFgf) and vascular endothelial growth factor (Vegf), and stained for nick end-labeling of DNA by the TUNEL method. Staining for distribution of AP in the arteriolar portion was reduced at both 1 and 3 weeks after irradiation with 20 Gy, but staining for DPP in the venular portion was unchanged, suggesting a close relationship between growth factors and injury of the arteriolar capillary portion. Fgf and Vegf proteins were present within the cytoplasm of the cardiomyocytes, or around capillaries, 1 h, 1 week and 3 weeks after irradiation. Many TUNEL-stained cardiomyocyte nuclei were observed at 1 h, but they had decreased markedly at 1 week and had almost disappeared by 3 weeks after irradiation. Thus Fgf and Vegf were induced concomitantly with the decrease in the staining for endothelial AP by 20 Gy X irradiation, which also caused microeffects as indicated by TUNEL staining of many nuclei at 1 h postirradiation.

Animals↗

PubMed2000

Effects of cationic charge on three-dimensional structures of intercalative complexes: structure of a bis-intercalated DNA complex solved by MAD phasing.

We characterize intercalative complexes as either "high charge" and "low charge". In low charge complexes, stacking interactions appear to dominate stability and structure. The dominance of stacking is evident in structures of daunomycin, nogalamycin, ethidium, and triostin A/echinomycin. By contrast in a DNA complex with the tetracationic metalloporphyrin CuTMPyP4 [copper (II) meso-tetra(N-methyl-4-pyridyl)porphyrin], electrostatic interactions appear to draw the porphyrin into the duplex interior, extending the DNA along its axis, and unstacking the DNA. Similarly, DNA complexes of tetracationic ditercalinium and tetracationic flexi-di show significant unstacking. Here we report x-ray structures of complexes of the tetracationic bis-intercalator D232 bound to DNA fragments d(CGTACG) and d(BrCGTABrCG). D232 is analogous to ditercalinium but with three methylene groups inserted between the piperidinium groups. The extension of the D232 linker allows it to sandwich four base pairs rather than two. In comparison to CuTMPyP4, flexi-di and ditercalinium, stacking interactions of D232 are significantly improved. We conclude that it is not sufficient to characterize intercalators simply by net charge. One anticipates strong electrostatic forces when cationic charge is focused to a small volume or region near DNA and so must consider the extent to which cationic charge is focused or distributed. In sum, ditercalinium, with a relatively short linker, focuses cationic charge more narrowly than does D232. So even though the net charges are equivalent, electrostatic charges are expected to be of greater structural significance in the ditercalinium complex than in the D232 complex.

Cations↗

PubMed2000

Isolation, characterization, and expression analysis of starch synthase IIa cDNA from wheat (Triticum aestivum L.).

We characterized three near-full-length putative homoeologous cDNA (Ss2a-1, Ss2a-2, and Ss2a-3) in wheat endosperm most similar to the maize zSSIIa. Polypeptide sequences deduced from three Ss2a cDNA clones share a 95% overall sequence similarity, and may thus have similar biochemical properties and may make identical contributions to starch biosynthesis in wheat endosperm. The accumulation of RNA transcripts corresponding to three Ss2a genes in developing endosperm varies among three cultivars studied, but usually peaks in young endosperm at about 10 days post anthesis (DPA). The polyclonal antibody for the SSIIa-1 recombinant protein strongly reacted to three previously identified granule-bound starch synthases of 100 to 115 kDa. The polyclonal antibody for the granule-bound starch synthases strongly reacted to the SSIIa-1 recombinant protein. Sequences of the N-terminal and an internal peptide of these three granule-bound starch synthases match well with those of three predicted mature SSIIa polypeptides. These granule-bound starch synthases may therefore be SSIIa polypeptides. The antibodies also recognized a group of three polypeptides with the same gel mobility as the three granule-bound starch synthases, a polypeptide of 90 kDa, and a group of three polypeptides of about 80 to 82 kDa. Thus, the wheat SSIIa may exist in several functional forms in the stroma of amyloplasts.

Amino Acid Sequence↗

PubMed2000

Changes in serum ammonia concentration in cirrhotic patients with Helicobacter pylori infection.

OBJECTIVE: To study whether liver cirrhosis associated with Helicobacter pylori (H. pylori) infection will induce increased serum ammonia and whether the peripheral serum ammonia reflects the level of portal vein serum ammonia. METHODS: Blood was taken from the portal vein and the cubital vein in cirrhotic patients with and without H. pylori infection and non-cirrhotic patients (splenic rupture) with and without H. pylori infection, and the serum ammonia was measured. RESULTS: The mean levels of serum ammonia in the group of cirrhotic patients with H. pylori infection were 167.82 +/- 8.97 mumol/L (portal vein) and 142.2 +/- 13.35 mumol/L (cubital vein). They were increased significantly as compared with cirrhotic patients without H. pylori infection (47.68 +/- 12.03 mumol/L portal vein and 37.23 +/- 7.04 mumol/L cubital vein), and also compared with the groups of splenic rupture patients with and without H. pylori infection (P < 0.01). There was no significant difference between the serum ammonia level of the cubital vein and portal vein (P > 0.05). CONCLUSIONS: H. pylori infection can induce an increase in serum ammonia in patients with liver dysfunction, and the peripheral serum ammonia measurement may replace the portal vein serum ammonia as a monitoring method. Eradication of H. pylori in cirrhotic patients may prevent hepatic encephalopathy (HE).

Ammonia↗

PubMed2000

[Intraoperative endoscopic sphincterotomy for common bile duct stones during laparoscopic cholecystectomy].

OBJECTIVE: To assess the effects of approach-intraoperative endoscopic sphincterotomy (IOES) for common bile duct (CBD) stones during laparoscopic cholecystectomy (LC). METHODS: Twenty-seven patients with secondary CBD stones were treated by IOES during LC. Therapeutic effects were evaluated on the basis of the cure rate, early complications, and days of hospitalization. RESULTS: IOES was successfully performed in 26 (96.30%) of 27 cases, and their CBD stones were cleared completely. Two cases (7.69%) were complicated by mild acute pancreatitis. CONCLUSION: IOES as an alternative to the treatment of CBD stones during LC is safe and avoids reoperation.

Adult↗

PubMed2000

[The epidemiological study of HIV infection among paid blood donors in one county of China].

OBJECTIVE: To observe the prevalence of HIV infection among paid blood donors and risk factors. METHODS: Selected the residents between 18 to 50 years old from two villages in one county of China in 1997. We observed the prevalence of HIV infection and keep following them to 2000. RESULTS: Nine hundred sixty-three residents were investigated and the prevalence of HIV was 1.8% (17/963). There were 88 paid blood donors among 963 interviewed subjects, the HIV prevalence was significantly higher (17.0% or 15/88) than those who didn't have the experiences of paid blood donations (0.2% or 2/875). Only one person out of 668 who were followed from 1997 to 2000 has converted to sero-positive, the incidence of HIV infection is 0.06 per 100 person-years. Nearly half of sero-negative and sero-positive people have the experiences of muscle injection. Among those with HIV-positive spouse, we found no one converted to sero-positive. The CD(4) count of those sero-positive people declined every year. HIV subtype and C2-V3 region sequence analysis were made for five HIV-positive people in 1998, the result indicated that all of them were infected with HIV-1 subtype B' and the genetic divergence rate was 2.4%. CONCLUSION: There was a high prevalence of HIV among paid blood donors in some regions, and paid blood (plasma) donations were the main risk factor for HIV infection. Although no evidence suggests that during the follow-up period the epidemic of HIV infection has spread locally, it still shall take measurements to prevent further transmission of HIV through paid blood donors.

Adolescent↗

PubMed2000

[Improving memory effects of eel oil capsule on memory obstruction of mice hurt by anisodine].

The effects of eel oil capsule on memory obstruction of mice were observed in step-down test, step-through test, Y-maze test and Japanese labyrinth test. The results indicated that the eel oil capsule treating group with dosages of 0.234, 0.702 and 2.106 g/kg markedly enhance the acquirement, strengthing and reappearance of memory, having the functions of improving brain memory hurt by Anisodine.

Animals↗

PubMed1999

Synthesis and biological evaluation of dihydrobenzofuran lignans and related compounds as potential antitumor agents that inhibit tubulin polymerization.

A series of 19 related dihydrobenzofuran lignans and benzofurans was obtained by a biomimetic reaction sequence involving oxidative dimerization of p-coumaric, caffeic, or ferulic acid methyl esters, followed by derivatization reactions. All compounds were evaluated for potential anticancer activity in an in vitro human disease-oriented tumor cell line screening panel that consisted of 60 human tumor cell lines arranged in nine subpanels, representing diverse histologies. Leukemia and breast cancer cell lines were relatively more sensitive to these agents than were the other cell lines. Compounds 2c and 2d, but especially 2b (methyl (E)-3-¿2-(3, 4-dihydroxyphenyl)-7-hydroxy-3-methoxycarbonyl-2, 3-dihydro-1-benzofuran-5-ylprop-2-enoate), the dimerization product of caffeic acid methyl ester, containing a 3',4'-dihydroxyphenyl moiety and a hydroxyl group in position 7 of the dihydrobenzofuran ring, showed promising activity. The average GI(50) value (the molar drug concentration required for 50% growth inhibition) of 2b was 0.3 microM. Against three breast cancer cell lines, 2b had a GI(50) value of <10 nM. Methylation, reduction of the double bond of the C(3)-side chain, reduction of the methoxycarbonyl functionalities to primary alcohols, or oxidation of the dihydrobenzofuran ring to a benzofuran system resulted in a decrease or loss of cytotoxic activity. Compound 2b inhibited mitosis at micromolar concentrations in cell culture through a relatively weak interaction at the colchicine binding site of tubulin. In vitro it inhibited tubulin polymerization by 50% at a concentration of 13 +/- 1 microM. The 2R, 3R-enantiomer of 2b was twice as active as the racemic mixture, while the 2S,3S-enantiomer had minimal activity as an inhibitor of tubulin polymerization. These dihydrobenzofuran lignans (2-phenyl-dihydrobenzofuran derivatives) constitute a new group of antimitotic and potential antitumor agents that inhibit tubulin polymerization.

Antineoplastic Agents↗

PubMed1999

Monoclonal antibodies that inhibit the transport function of the 190-kDa multidrug resistance protein, MRP. Localization of their epitopes to the nucleotide-binding domains of the protein.

Multidrug resistance in tumor cells is often accompanied by overexpression of multidrug resistance protein (MRP), a 190-kDa transmembrane protein that belongs to the ATP-binding cassette superfamily of transport proteins. MRP mediates ATP-dependent transport of a variety of conjugated organic anions and can also transport several unmodified xenobiotics in a glutathione-dependent manner. To facilitate structure-function studies of MRP, we have generated a panel of MRP-specific monoclonal antibodies (mAbs). Four of these mAbs, QCRL-2, -3, -4, and -6, bind intracellular conformation-dependent epitopes, and we have shown that they can inhibit the transport of several MRP substrates. Binding competition and immunoprecipitation assays indicated that mAbs QCRL-4 and -6 probably recognize the same detergent-sensitive epitope in MRP, whereas mAbs QCRL-2, -3, and -4 each bind distinct, non-overlapping epitopes. Fab fragments inhibit transport as effectively as the intact mAbs, suggesting that inhibition results from direct interactions of the mAbs with MRP. Immunodot blot and immunoprecipitation analyses revealed that the minimal regions of MRP sufficient for full reactivity of mAbs QCRL-2 and -3 are amino acids 617-858 and 617-932, respectively, which encompass the NH2-proximal nucleotide-binding domain (NBD). In contrast, the epitope bound by mAb QCRL-4 localized to amino acids 1294-1531, a region that contains the COOH-proximal NBD. However, none of the mAbs inhibited photolabeling of intact MRP with 8-azido-[alpha-32P]ATP. This suggests that rather than preventing nucleotide binding, the mAbs inhibit transport by interfering with substrate binding or by trapping MRP in a conformation that does not allow transport to occur. Our results also demonstrate for the first time that the NBDs of MRP can be expressed as soluble polypeptides that retain a native conformation.

ATP-Binding Cassette Transporters↗

PubMed1999

Activation of Rb and decline in androgen receptor protein precede retinoic acid-induced apoptosis in androgen-dependent LNCaP cells and their androgen-independent derivative.

Androgen ablation-induced prostate cancer regression is transient and ends with the regrowth of androgen-independent (AI) tumors. To mimic this evolution in culture, we chronically deprived an androgen-dependent (AD) prostate cancer cell line (LNCaP) of androgen, generating an AI derivative which retained limited hormone proliferative responsiveness and a barely detectable prostate-specific antigen (PSA) mRNA level. While the cytokeratin 8 (CK8) level was low, the androgen receptor (AR) protein in AI cells was on average tenfold greater than in AD cells. When challenged for susceptibility to undergo apoptosis, the AI cells were more resistant than AD cells to all-trans retinoic acid (tRA) and two chemotherapeutic agents, Taxol and Adriamycin, requiring higher doses and longer periods of treatment to achieve similar effects. Compared to AD cells, the partially apoptosis-resistant AI cells expressed four times more Bcl-2 protein and undetectable levels of p21/WAF1. Induction of apoptosis by tRA in both cell types did not affect their expression but was preceded by the activation of Rb and a pronounced reduction of AR protein level. The kinetics of the Rb activation and AR downmodulation in both cell types matched their tRA sensitivity, suggesting that these events may be required for tRA-induced apoptosis. The results show that the apoptotic pathway in AI cells, although more difficult to induce, is not irrevocably lost and that targeted reduction of the AR protein level with retinoids in combination with androgen ablation therapy may prolong remissions in advanced prostate cancer patients.

Antigens, Surface↗