Additionally, fibroblasts from ATF3 knock-out mice were partially protected from UV-induced apoptosis Even plants Arabidopsis thaliana treated with doxycycline displayed marked growth retardation, while on a molecular level they presented with mitochondrial dysfunction and activated UPRmt The recruitment of DNA repair factors was shown to be enhanced by cyclin D1, but not the related cyclin D1b isoform PC-3 cells died through apoptosis as evidenced by the enzyme-linked immunosorbent assay measuring mono- and oligonucleosomes in the lysates of apoptotic cells Fig.
The empty vector was used to generate negative controls. Overproduction of p52 has been observed in several solid tumors including breast and prostate cancers 11 Among our microarray analysis results, the expression of p57Kip2 decreased over 2-fold in all ages of 12T-7f mice, compared with that of normal mice, which was confirmed by quantitative real-time reverse transcription—PCR RT-PCR; data not shown.
The clonogenic survival fraction was set to the cell plating efficiency. Enzalutamide binds to the AR with greater affinity than bicalutamide and inhibits its nuclear translocation and expression of its target genes 3.
Complete protease inhibitor mixture Roche Applied Science was added to lysis buffer before use. Cells were arrested by starvation in 0. Furthermore, stress conditions exposure to staurosporine and hypoxia induced by sodium azide caused significant increase in ATF3 expression and induced apoptosis, whereas knockdown of KLF6 by small interference RNA blocked the increase of ATF3 as well as the induction of apoptosis in these conditions.
At the end of the experiment, cells were fixed with methanol, stained with crystal violet, and the numbers of colonies were counted. Cell culture and materials. Next to their vital metabolic role, they are also involved in many other important processes of the cell, such as calcium homeostasis, apoptosis and intra-cellular signaling 1.
Besides growth inhibition, KLF6 has also been shown to induce apoptosis in non-small cell lung cancer cells 1.
Our results show that decreased expression of p57Kip2 occurs frequently in human prostate cancer, even in the precursor PIN stage. After washing thrice for 10 min each in TBS-T, incubation was performed for 1 h with the secondary horseradish peroxidase—conjugated goat anti-rabbit or goat anti-mouse antibodies.
Anti-KLF6 monoclonal antibody was purchased from Invitrogen. Results p57Kip2 expression is decreased in human prostate cancer.
Remarkably, this mouse prostate cancer is pathologically identical to human prostate adenocarcinoma. Although cells are initially dependent on mitogenic stimuli for passage through G1, progression beyond the restriction point R1 results in autonomy of the cycle 2.
Antibodies against AR-V7 splice variant were kindly provided by Dr. The experiments were done in triplicate wells. To identify genes involved in tumorigenesis, we performed cDNA microarrays to directly compare 2-week-old, 4-week-old, 6-week-old, week-old, and week-old 12T-7f dorsolateral prostate tumors with 2-week-old normal mouse prostate tissues standard.
Protein lysates were prepared, and immunoblot experiments were performed as previously described Staurosporine was dissolved in dimethyl sulfoxide DMSO at a concentration of 1 mm. Additionally, we show that overexpression of p57Kip2 decreases cell proliferation and tumorigenesis. Successive rounds of infection over 5 d were used.
To understand p57Kip2 expression in human prostate cancer, we performed immunohistochemical staining in human prostate cancer tissues. After 14 days, colonies were stained with crystal violet solution and counted. S1Aconsistent with a previous report To understand the mechanism of KLF6-induced apoptosis, we employed microarray gene expression analysis and identified ATF3 as one of the target genes regulated by KLF6.
Furthermore, the prostates of p57Kip2 knockout mice developed prostatic intraepithelial neoplasia and adenocarcinoma.
PDF Abstract Therapy resistance and poor outcome in prostate cancer is associated with increased expression of cyclin D1.
The functional significance of p processing has been confirmed by genetic evidence from humans and mice Numerous mitogenic and quiescence-inducing stimuli must converge at certain points to allow a coordinated cellular response that typifies the normal cell cycle.
Final Gleason scores varied from 5 to 9 Gleason scores of 5 for 4 patients, 6 for 19 patients, 7 for 17 patients, 8 for 1 patient, and 9 for 1 patient.
Considering the versatile functions of mitochondria, it is no surprise that dysfunction of these organelles can lead to manifestation of different diseases affecting various tissues and with symptoms of ranging severity that can even be life-threatening 2.
The antibody p57 clone H91, Santa Cruz; 1: Viable cells were then counted using a hemacytometer, with viability determined by trypan blue exclusion. The processing of p to p52 is a tightly controlled event in many cells and tissues 6 — 9.
On the other hand, it was shown that ATF3 overexpression promoted invasiveness of prostate tumor cells in vitro and significantly enhanced spontaneous lung metastasis without affecting primary tumorigenicity in a severe combined immunodeficient mouse model Indeed, similar to RNAi-mediated down-regulation of the mitochondrial translational machinery, treatment with doxycycline—an antibiotic belonging to the family of tetracyclines—induced mitonuclear protein imbalance, impaired mitochondrial respiration, activated UPRmt and dose-dependently extended the nematode lifespan 5.trial in prostate cancer patients.
The biological role played in tumor growth is presently unknown. In this report we Methods: A bladder cell line was engineered to express a doxycycline (dox) regulated shRNA against PSCA. To shed light on the PSCA biological role in tumor growth, microarray analysis was carried out as a function of PSCA.
Selenium-Binding Protein 1 (SBP1, SELENBP1, hSP56) is a selenium-associated protein shown to be at lower levels in tumors, and its lower levels are frequently predictive of a poor clinical outcome. Distinguishing indolent from aggressive prostate cancer is a major challenge in disease management.
Mutations of KLF6 gene have been identified in clinical prostate cancer samples (2, 17), suggesting a tumor suppressor role for KLF6 in prostate cancer. Indeed, decreased KLF6 expression has been shown to correlate with clinical outcome in prostate cancer patients (23).
Nov 01, · Strikingly, a reanalysis of microarray transcriptome profiles of human bladder cancer cell line RT exposed to doxycycline showed a 10% change in global gene expression, confirming the molecular basis for the widespread adverse effects of this treatment (Fig.
1). A doxycycline-inducible urokinase receptor (uPAR) upregulates uPAR activities including resistance to anoikis in human prostate cancer cell lines. Fife RS, Sledge GW Jr, Roth BJ, Proctor C () Effects of doxycycline on human prostate cancer cells in vitro.
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