Real-time monitoring of active caspase 3 during AFB1 induced apoptosis based on SERS-fluorescent dual mode signals

Background/Aims: MiR-19b has been reported to be involved in several malignancies, but its role in multiple myeloma (MM) is still unknown. The objective of this study was to explore the biological mechanism of miR-19b in the progression of MM. Methods: First, we performed real-time polymerase chain reaction (PCR) and Western blot to study the expression of miR-19b, tuberous sclerosis 1 (TSC1), and caspase-3 in different groups. MTT assay was performed to explore the effect of miR-19b on survival and apoptosis of cancer stem cells (CSCs). Computation analysis and luciferase assay were utilized to confirm the interaction between miR-19b and TSC1. Results: A total of 38 participants comprising 20 subjects with MM and 18 healthy subjects as normal controls were enrolled in our study. Real-time PCR showed dramatic upregulation of miR-19b, but TSC1 was evidently suppressed in the MM group. MiR-19b overexpression substantially promoted clonogenicity and cell viability, and further inhibited apoptosis of CSCs in vitro. Furthermore, miR-19b overexpression downregulated the expression of caspase-3, which induced apoptosis. Using in silico analysis, we identified that TSC1 might be a direct downstream target of miR-19b, and this was further confirmed by luciferase assay showing that miR-19b apparently reduced the luciferase activity of wild-type TSC1 3´-UTR, but not that of mutant TSC1 3´-UTR. There was also evident decrease in TSC1 mRNA and protein in CSCs following introduction of miR-19b. Interestingly, reintroduction of TSC1 abolished the miR-19b-induced proliferation promotion and apoptosis inhibition in CSCs. Conclusion: These findings collectively suggest that miR-19b promotes cell survival and suppresses apoptosis of MM CSCs via targeting TSC1 directly, indicating that miR-19b may serve as a potential and novel therapeutic target of MM based on miRNA expression.

Download Full-text

Porphyra-334, a mycosporine-like amino acid, attenuates UV-induced apoptosis in HaCaT cells

Acta Pharmaceutica ◽

10.1515/acph-2017-0015 ◽

2017 ◽

Vol 67 (2) ◽

pp. 257-264 ◽

Cited By ~ 6

Author(s):

Sung-Suk Suh ◽

Se Kyung Oh ◽

Sung Gu Lee ◽

Il-Chan Kim ◽

Sanghee Kim

Keyword(s):

Caspase 3 ◽

Biological Activities ◽

Inhibitory Effect ◽

In Vitro Assays ◽

Control Group ◽

Hacat Cells ◽

Dramatic Decrease ◽

Induced Apoptosis ◽

Active Caspase

Abstract The main aim of the current research was to study the effect of porphyra-334, one of mycosporine-like amino acids (MAAs), well known as UV-absorbing compounds, on UVinduced apoptosis in human immortalized keratinocyte (HaCaT) cells. Due to their UV-screening capacity and ability to prevent UV-induced DNA damage, MAAs have recently attracted considerable attention in both industry and research in pharmacology. Herein, human HaCaT cells were used to determine the biological activities of porphyra- 334 by various in vitro assays, including proliferation, apoptosis and Western blot assays. The proliferation rate of UV-irradiated HaCaT cells was significantly decreased compared to the control group. Pretreatment with porphyra- 334 markedly attenuated the inhibitory effect of UV and induced a dramatic decrease in the apoptotic rate. Expression of active caspase-3 protein was increased in response to UV irradiation, while caspase-3 levels were similar between cells treated with porphyra-334 and the non-irradiated control group. Taken together, our data suggest that porphyra-334 inhibits UV-induced apoptosis in HaCaT cells through attenuation of the caspase pathway.

Download Full-text

Activatable Ferritin Nanocomplex for Real-Time Monitoring of Caspase-3 Activation during Photodynamic Therapy

ACS Applied Materials & Interfaces ◽

10.1021/acsami.5b07316 ◽

2015 ◽

Vol 7 (41) ◽

pp. 23248-23256 ◽

Cited By ~ 29

Author(s):

Jingjing Wang ◽

Liwen Zhang ◽

Minglong Chen ◽

Shi Gao ◽

Lei Zhu

Keyword(s):

Photodynamic Therapy ◽

Real Time ◽

Caspase 3 ◽

Real Time Monitoring

Download Full-text

Real-time monitoring of caspase-3/8 activity by self-assembling nanofiber probes in living cells

Chemical Communications ◽

10.1039/d0cc07821b ◽

2021 ◽

Author(s):

Li-Song Zhang ◽

Hong-Lei Xu ◽

Ying Xia ◽

Jian-Peng Bi ◽

Chuan-Zeng Zhang ◽

...

Keyword(s):

Real Time ◽

Caspase 3 ◽

Living Cells ◽

Real Time Monitoring ◽

Self Assembling

Supramolecular self-assembling nanofiber probes Nap-GFFpYDEVD-AFC/Nap-GFFpYIETD-AFC for real-time monitoring of caspase-3/8 activity in living cells.

Download Full-text

NF-κB-mediated IAP expression induces resistance of intestinal epithelial cells to apoptosis after polyamine depletion

AJP Cell Physiology ◽

10.1152/ajpcell.00480.2003 ◽

2004 ◽

Vol 286 (5) ◽

pp. C1009-C1018 ◽

Cited By ~ 67

Author(s):

Tongtong Zou ◽

Jaladanki N. Rao ◽

Xin Guo ◽

Lan Liu ◽

Huifang M. Zhang ◽

...

Keyword(s):

Epithelial Cells ◽

Caspase 3 ◽

Intestinal Epithelial Cells ◽

Specific Inhibitor ◽

Inhibitory Effect ◽

Intestinal Epithelial ◽

Tnf Α ◽

Induced Apoptosis ◽

Necrosis Factor ◽

Active Caspase

Apoptosis plays a crucial role in maintenance of intestinal epithelial integrity and is highly regulated by numerous factors, including cellular polyamines. We recently showed that polyamines regulate nuclear factor (NF)-κB activity in normal intestinal epithelial (IEC-6) cells and that polyamine depletion activates NF-κB and promotes resistance to apoptosis. The current study went further to determine whether the inhibitors of apoptosis (IAP) family of proteins, c-IAP2 and XIAP, are downstream targets of activated NF-κB and play a role in antiapoptotic activity of polyamine depletion in IEC-6 cells. Depletion of cellular polyamines by α-difluoromethylornithine not only activated NF-κB activity but also increased expression of c-IAP2 and XIAP. Specific inhibition of NF-κB by the recombinant adenoviral vector containing IκBα superrepressor (Ad Iκ BSR) prevented the induction of c-IAP2 and XIAP in polyamine-deficient cells. Decreased levels of c-IAP2 and XIAP proteins by inactivation of NF-κB through Ad Iκ BSR infection or treatment with the specific inhibitor Smac also overcame the resistance of polyamine-depleted cells to apoptosis induced by the combination of tumor necrosis factor (TNF)-α and cycloheximide (CHX). Although polyamine depletion did not alter levels of procaspase-3 protein, it inhibited formation of the active caspase-3. Decreased levels of c-IAP2 and XIAP by Smac prevented the inhibitory effect of polyamine depletion on the cleavage of procaspase-3 to the active caspase-3. These results indicate that polyamine depletion increases expression of c-IAP2 and XIAP by activating NF-κB in intestinal epithelial cells. Increased c-IAP2 and XIAP after polyamine depletion induce the resistance to TNF-α/CHX-induced apoptosis, at least partially, through inhibition of the caspase-3 activity.

Download Full-text

Fusion-Induced Apoptosis Contributes to Thymocyte Depletion by a Pathogenic Human Immunodeficiency Virus Type 1 Envelope in the Human Thymus

Journal of Virology ◽

10.1128/jvi.01382-06 ◽

2006 ◽

Vol 80 (22) ◽

pp. 11019-11030 ◽

Cited By ~ 21

Author(s):

Eric G. Meissner ◽

Liguo Zhang ◽

S. Jiang ◽

Lishan Su

Keyword(s):

Human Immunodeficiency Virus ◽

Human Immunodeficiency Virus Type ◽

Virus Type ◽

Caspase 3 ◽

Fusion Inhibitors ◽

Immunodeficiency Virus ◽

Induced Apoptosis ◽

Hiv 1 ◽

Active Caspase

ABSTRACT The mechanisms of CD4+ T-cell depletion during human immunodeficiency virus type 1 (HIV-1) infection remain incompletely characterized. Of particular importance is how CD4+ T cells are depleted within the lymphoid organs, including the lymph nodes and thymus. Herein we characterize the pathogenic mechanisms of an envelope from a rapid progressor (R3A Env) in the NL4-3 backbone (NL4-R3A) which is able to efficiently replicate and deplete CD4+ thymocytes in the human fetal-thymus organ culture (HF-TOC). We demonstrate that uninterrupted replication is required for continual thymocyte depletion. During depletion, NL4-R3A induces an increase in thymocytes which uptake 7AAD, a marker of cell death, and which express active caspase-3, a marker of apoptosis. While 7AAD uptake is observed predominantly in uninfected thymocytes (p24−), active caspase-3 is expressed in both infected (p24+) and uninfected thymocytes (p24−). When added to HF-TOC with ongoing infection, the protease inhibitor saquinavir efficiently suppresses NL4-R3A replication. In contrast, the fusion inhibitors T20 and C34 allow for sustained HIV-1 production. Interestingly, T20 and C34 effectively prevent thymocyte depletion in spite of this sustained replication. Apoptosis of both p24− and p24+ thymocytes appears to be envelope fusion dependent, as T20, but not saquinavir, is capable of reducing thymocyte apoptosis. Together, our data support a model whereby pathogenic envelope-dependent fusion contributes to thymocyte depletion in HIV-1-infected thymus, correlated with induction of apoptosis in both p24+ and p24− thymocytes.

Download Full-text

Erythropoietin Prevents Dialysis Fluid-Induced Apoptosis of Mesothelial Cells

Peritoneal Dialysis International ◽

10.1177/089686080802800618 ◽

2008 ◽

Vol 28 (6) ◽

pp. 648-654 ◽

Cited By ~ 5

Author(s):

Marina Vorobiov ◽

Myriam Malki ◽

Alla Shnaider ◽

Ana Basok ◽

Boris Rogachev ◽

...

Keyword(s):

Caspase 3 ◽

Specific Inhibitor ◽

Mesothelial Cells ◽

Short Exposure ◽

Peritoneal Fibrosis ◽

Peritoneal Mesothelial Cells ◽

Visceral Peritoneum ◽

Induced Apoptosis ◽

Active Caspase

Background In peritoneal dialysis (PD)-treated patients, denudation of the mesothelium correlates with peritoneal fibrosis and vascular changes. Since recombinant human erythropoietin (rHuEPO) induces a range of cytoprotective cellular responses, rHuEPO treatment may reduce PD fluid (PDF)-induced damage. Methods To investigate the antiapoptotic effect and mechanism of rHuEPO in peritoneal mesothelial cells (PMCs), isolated mice PMCs were used for in vitro characterization of rHuEPO effects. To confirm the in vitro effects, active caspase-3 was analyzed in imprints of liver visceral peritoneum of mice pretreated overnight with rHuEPO (5000 U/kg intraperitoneally) and exposed to PDF (Dianeal 4.25%; Baxter Healthcare, Deerfield, Illinois, USA) for 4 hours. Results Mouse PMCs expressed EPO-receptor mRNA and protein. Short exposure to rHuEPO (5 U/mL) induced phosphorylation of JAK2, STAT5, and ERK1/2. PMCs pretreated for 1 hour with rHuEPO showed reduced PDF-induced caspase-3 activation (49.6%) and DNA fragmentation (38.4%) in comparison to cells treated by PDF alone ( p < 0.05). rHuEPO treatment induced an increase in ERK1/2 phosphorylation and reduced levels of PDF-induced phospho-P38. PD98059, a specific inhibitor of ERK activation, fully blocked the protective effect of rHuEPO. In mice, rHuEPO reduced the apoptotic effect of PDF, as assessed by the level of active caspase-3. Conclusions Our study presents new insights into clinical use of rHuEPO in the setting of PD. We found that rHuEPO provides ERK1/2-dependent protection to PMCs from PDF-induced apoptosis. The use of rHuEPO, or any of its new derivatives that do not stimulate erythropoiesis, should be considered for peritoneal preservation.

Download Full-text