Silencing of Bcl-2 expression by small interfering RNA induces autophagic cell death in MCF-7 breast cancer cells.

Abstract:

:Apoptosis (programmed cell death type I) and autophagy (type II) are crucial mechanisms regulating cell death and homeostasis. The Bcl-2 proto-oncogene is overexpressed in 50-70% of breast cancers, potentially leading to resistance to chemotherapy, radiation and hormone therapy-induced apoptosis. Here, we investigated the role of Bcl-2 in autophagy in breast cancer cells. Silencing of Bcl-2 by siRNA in MCF-7 breast cancer cells downregulated Bcl-2 protein levels (>85%) and led to inhibition of cell growth (71%) colony formation (79%), and cell death (up to 55%) by autophagy but not apoptosis. Induction of autophagy was demonstrated by acridine orange staining, electron microscopy and an accumulation of GFP-LC3-II in autophagosomal membranes in MCF-7 cells transfected with GFP-LC-3(GFP-ATG8). Silencing of Bcl-2 by siRNA also led to induction of LC-3-II, a hallmark of autophagy, ATG5 and Beclin-1 autophagy promoting proteins. Knockdown of ATG5 significantly inhibited Bcl-2 siRNA-induced LC3-II expression, the number of GFP-LC3-II-labeled autophagosome positive cells and autophagic cell death (p < 0.05). Furthermore, doxorubicin at a high dose (IC(95), 1 microM) induced apoptosis but at a low dose (IC(50), 0.07 microM) induced only autophagy and Beclin-1 expression. When combined with Bcl-2 siRNA, doxorubicin (IC(50)) enhanced autophagy as indicated by the increased number cells with GFP-LC3-II-stained autophagosomes (punctuated pattern positive). These results provided the first evidence that targeted silencing of Bcl-2 induces autophagic cell death in MCF-7 breast cancer cells and that Bcl-2 siRNA may be used as a therapeutic strategy alone or in combination with chemotherapy in breast cancer cells that overexpress Bcl-2.

journal_name

Autophagy

journal_title

Autophagy

authors

Akar U,Chaves-Reyez A,Barria M,Tari A,Sanguino A,Kondo Y,Kondo S,Arun B,Lopez-Berestein G,Ozpolat B

doi

10.4161/auto.6083

subject

Has Abstract

pub_date

2008-07-01 00:00:00

pages

669-79

issue

5

eissn

1554-8627

issn

1554-8635

pii

6083

journal_volume

4

pub_type

杂志文章
  • microRNA 30A promotes autophagy in response to cancer therapy.

    abstract::microRNAs (miRNAs) are a class of small regulatory RNAs that regulate gene expression at the post-transcriptional level. miRNAs play important roles in the regulation of development, growth, and metastasis of cancer, and in determining the response of tumor cells to anticancer therapy. In recent years, they have also ...

    journal_title:Autophagy

    pub_type: 杂志文章

    doi:10.4161/auto.20053

    authors: Yu Y,Cao L,Yang L,Kang R,Lotze M,Tang D

    更新日期:2012-05-01 00:00:00

  • Therapeutic effects of remediating autophagy failure in a mouse model of Alzheimer disease by enhancing lysosomal proteolysis.

    abstract::The extensive autophagic-lysosomal pathology in Alzheimer disease (AD) brain has revealed a major defect: in the proteolytic clearance of autophagy substrates. Autophagy failure contributes on several levels to AD pathogenesis and has become an important therapeutic target for AD and other neurodegenerative diseases. ...

    journal_title:Autophagy

    pub_type: 杂志文章

    doi:10.4161/auto.7.7.15596

    authors: Yang DS,Stavrides P,Mohan PS,Kaushik S,Kumar A,Ohno M,Schmidt SD,Wesson DW,Bandyopadhyay U,Jiang Y,Pawlik M,Peterhoff CM,Yang AJ,Wilson DA,St George-Hyslop P,Westaway D,Mathews PM,Levy E,Cuervo AM,Nixon RA

    更新日期:2011-07-01 00:00:00

  • Macrophages target Salmonella by Lc3-associated phagocytosis in a systemic infection model.

    abstract::Innate immune defense against intracellular pathogens, like Salmonella, relies heavily on the autophagy machinery of the host. This response is studied intensively in epithelial cells, the target of Salmonella during gastrointestinal infections. However, little is known of the role that autophagy plays in macrophages,...

    journal_title:Autophagy

    pub_type: 杂志文章

    doi:10.1080/15548627.2019.1569297

    authors: Masud S,Prajsnar TK,Torraca V,Lamers GEM,Benning M,Van Der Vaart M,Meijer AH

    更新日期:2019-05-01 00:00:00

  • Methods to monitor autophagy in H. pylori vacuolating cytotoxin A (VacA)-treated cells.

    abstract::Helicobacter pylori is a gram negative pathogen that infects at least half of the world's population and is associated not only with gastric cancer but also with other diseases such as gastritis and peptic ulcers. Indeed, H. pylori is considered the single most important risk factor for the development of gastric canc...

    journal_title:Autophagy

    pub_type: 杂志文章,评审

    doi:10.4161/auto.6.1.10222

    authors: Raju D,Jones NL

    更新日期:2010-01-01 00:00:00

  • Role of actin in shaping autophagosomes.

    abstract::One of the main unanswered questions regarding the early steps of macroautophagy/autophagy is the mechanism of membrane-modeling events required for autophagosome formation. Three independent studies have recently revealed an actin cytoskeleton involvement in this process, providing significant details regarding the r...

    journal_title:Autophagy

    pub_type: 杂志文章

    doi:10.1080/15548627.2016.1236877

    authors: Zientara-Rytter K,Subramani S

    更新日期:2016-12-01 00:00:00

  • Upregulation of SQSTM1/p62 contributes to nickel-induced malignant transformation of human bronchial epithelial cells.

    abstract::Chronic lung inflammation is accepted as being associated with the development of lung cancer caused by nickel exposure. Therefore, identifying the molecular mechanisms that lead to a nickel-induced sustained inflammatory microenvironment that causes transformation of human bronchial epithelial cells is of high signif...

    journal_title:Autophagy

    pub_type: 杂志文章

    doi:10.1080/15548627.2016.1196313

    authors: Huang H,Zhu J,Li Y,Zhang L,Gu J,Xie Q,Jin H,Che X,Li J,Huang C,Chen LC,Lyu J,Gao J,Huang C

    更新日期:2016-10-02 00:00:00

  • The PI 3-kinase regulator Vps15 is required for autophagic clearance of protein aggregates.

    abstract::Autophagy is involved in cellular clearance of aggregate-prone proteins, thereby having a cytoprotective function. Studies in yeast have shown that the PI 3-kinase Vps34 and its regulatory protein kinase Vps15 are important for autophagy, but the possible involvement of these proteins in autophagy in a multicellular a...

    journal_title:Autophagy

    pub_type: 杂志文章

    doi:10.4161/auto.5829

    authors: Lindmo K,Brech A,Finley KD,Gaumer S,Contamine D,Rusten TE,Stenmark H

    更新日期:2008-05-01 00:00:00

  • Autophagy, Inflammation, and Metabolism (AIM) Center in its second year.

    abstract::The NIH-funded center for autophagy research named Autophagy, Inflammation, and Metabolism (AIM) Center of Biomedical Research Excellence, located at the University of New Mexico Health Science Center is now completing its second year as a working center with a mission to promote autophagy research locally, nationally...

    journal_title:Autophagy

    pub_type: 历史文章,杂志文章

    doi:10.1080/15548627.2019.1634444

    authors: Deretic V,Prossnitz E,Burge M,Campen MJ,Cannon J,Liu KJ,Liu M,Hall P,Sklar LA,Allers L,Mariscal L,Garcia SA,Weaver J,Baehrecke EH,Behrends C,Cecconi F,Codogno P,Chen GC,Elazar Z,Eskelinen EL,Fourie B,Gozuacik D

    更新日期:2019-10-01 00:00:00

  • DAMPs and autophagy: cellular adaptation to injury and unscheduled cell death.

    abstract::Autophagy is a lysosome-mediated catabolic process involving the degradation of intracellular contents (e.g., proteins and organelles) as well as invading microbes (e.g., parasites, bacteria and viruses). Multiple forms of cellular stress can stimulate this pathway, including nutritional imbalances, oxygen deprivation...

    journal_title:Autophagy

    pub_type: 杂志文章,评审

    doi:10.4161/auto.23691

    authors: Zhang Q,Kang R,Zeh HJ 3rd,Lotze MT,Tang D

    更新日期:2013-04-01 00:00:00

  • Lysosomal calcium regulates autophagy.

    abstract::Recent evidence has indicated that the lysosome is able to act as a signaling organelle that senses nutrient availability and generates an adaptive response that is important for cellular homeostasis. We recently discovered another example of lysosomal signaling where lysosomal calcium release activates the master aut...

    journal_title:Autophagy

    pub_type: 杂志文章,评审

    doi:10.1080/15548627.2015.1047130

    authors: Medina DL,Ballabio A

    更新日期:2015-01-01 00:00:00

  • 4th International Symposium on Autophagy: exploiting the frontiers of autophagy research.

    abstract::The 4th International Symposium on Autophagy was held in Mishima, a small town between Tokyo and Kyoto, October 1-5, 2006 (http://isa4th.umin.jp/). The meeting was organized by the group of Eiki Kominami. Approximately 150 participants took part in this well-organized meeting in the spacious and comfortable Toray Conf...

    journal_title:Autophagy

    pub_type:

    doi:10.4161/auto.3654

    authors: Eskelinen EL,Deretic V,Neufeld T,Levine B,Cuervo AM

    更新日期:2007-03-01 00:00:00

  • Autophagy-based unconventional secretion of HMGB1 by keratinocytes plays a pivotal role in psoriatic skin inflammation.

    abstract::The precise mechanism through which macroautophagy/autophagy affects psoriasis is poorly understood. Here, we found that keratinocyte (KC) autophagy, which was positively correlated with psoriatic severity in patients and mouse models and could be inhibited by mitogen-activated protein kinase (MAPK) family inactivatio...

    journal_title:Autophagy

    pub_type: 杂志文章

    doi:10.1080/15548627.2020.1725381

    authors: Wang Z,Zhou H,Zheng H,Zhou X,Shen G,Teng X,Liu X,Zhang J,Wei X,Hu Z,Zeng F,Hu Y,Hu J,Wang X,Chen S,Cheng J,Zhang C,Gui Y,Zou S,Hao Y,Zhao Q,Wu W,Zhou Y,Cui K,Huang N,Wei Y,Li W,Li J

    更新日期:2020-02-16 00:00:00

  • Context effect: microRNA-10b in cancer cell proliferation, spread and death.

    abstract::Single microRNA (miRNA) can regulate expression of several or multiple principal targets in a specific microenvironment. In different cellular contexts, the same miRNA may exhibit diverse functions, depending on the repertoire and stoichiometry of its direct mRNA targets. For instance, in breast cancer, microRNA-10b (...

    journal_title:Autophagy

    pub_type: 杂志文章

    doi:10.4161/auto.7.11.17371

    authors: Gabriely G,Teplyuk NM,Krichevsky AM

    更新日期:2011-11-01 00:00:00

  • AMPK connects energy stress to PIK3C3/VPS34 regulation.

    abstract::The class III phosphatidylinositol (PtdIns)-3 kinase, PIK3C3/VPS34, forms multiple complexes and regulates a variety of cellular functions, especially in intracellular vesicle trafficking and autophagy. Even though PtdIns3P, the product of PIK3C3, is thought to be a critical membrane marker for the autophagosome, it i...

    journal_title:Autophagy

    pub_type: 杂志文章

    doi:10.4161/auto.24877

    authors: Kim J,Guan KL

    更新日期:2013-07-01 00:00:00

  • Macroautophagy: protector in the diabetes drama?

    abstract::Macroautophagy ("autophagy") is regulated by the same insulin-amino acid-mTOR signaling pathway that controls protein synthesis. Although the literature does not so far include any direct studies confirming this, we expect autophagy to increase during insulin resistance. We discuss the possibility that this may be a u...

    journal_title:Autophagy

    pub_type: 杂志文章

    doi:10.4161/auto.4449

    authors: Meijer AJ,Codogno P

    更新日期:2007-09-01 00:00:00

  • The interplay between PRKCI/PKCλ/ι, SQSTM1/p62, and autophagy orchestrates the oxidative metabolic response that drives liver cancer.

    abstract::Hepatocellular carcinoma (HCC) is the consequence of chronic liver damage caused by the excessive generation of reactive oxygen species (ROS). To mitigate the deleterious effects of ROS, cells activate the transcription factor NFE2L2/NRF2, which is constitutively degraded through its partner KEAP1. The inactivation of...

    journal_title:Autophagy

    pub_type: 评论,杂志文章

    doi:10.1080/15548627.2020.1797290

    authors: Moscat J,Diaz-Meco MT

    更新日期:2020-10-01 00:00:00

  • Autophagy defects in Lafora disease: cause or consequence?

    abstract::Lafora disease (LD) is an inherited and fatal form of neurodegenerative disorder characterized by the presence of an abnormal form of glycogen inclusions, called Lafora bodies, in neurons and other tissues. While Lafora bodies have been thought to underlie the neuropathology in LD, the specific process by which these ...

    journal_title:Autophagy

    pub_type: 杂志文章,评审

    doi:10.4161/auto.8.2.19010

    authors: Puri R,Ganesh S

    更新日期:2012-02-01 00:00:00

  • Autophagosome formation: Where the secretory and autophagy pathways meet.

    abstract::The upregulation of autophagosome formation in response to nutrient deprivation requires significant intracellular membrane rearrangements that are poorly understood. Recent findings have implicated COPII-coated vesicles, well known as ER-Golgi cargo transport carriers, as key players in macroautophagy. The role of CO...

    journal_title:Autophagy

    pub_type: 杂志文章,评审

    doi:10.1080/15548627.2017.1287657

    authors: Wang J,Davis S,Zhu M,Miller EA,Ferro-Novick S

    更新日期:2017-05-04 00:00:00

  • Role of ULK-FIP200 complex in mammalian autophagy: FIP200, a counterpart of yeast Atg17?

    abstract::The yeast serine threonine kinase Atg1 appears to be a key regulator of autophagy and its kinase activity is crucial for autophagy induction. Recent reports have indicated that a mammalian Atg1 homolog, UNC-51-like kinase (ULK) 1, is required for autophagy. We found that ULK1 localizes to the autophagic isolation memb...

    journal_title:Autophagy

    pub_type: 杂志文章

    doi:10.4161/auto.5.1.7180

    authors: Hara T,Mizushima N

    更新日期:2009-01-01 00:00:00

  • SIRT5 regulation of ammonia-induced autophagy and mitophagy.

    abstract::In liver the mitochondrial sirtuin, SIRT5, controls ammonia detoxification by regulating CPS1, the first enzyme of the urea cycle. However, while SIRT5 is ubiquitously expressed, urea cycle and CPS1 are only present in the liver and, to a minor extent, in the kidney. To address the possibility that SIRT5 is involved i...

    journal_title:Autophagy

    pub_type: 杂志文章

    doi:10.1080/15548627.2015.1009778

    authors: Polletta L,Vernucci E,Carnevale I,Arcangeli T,Rotili D,Palmerio S,Steegborn C,Nowak T,Schutkowski M,Pellegrini L,Sansone L,Villanova L,Runci A,Pucci B,Morgante E,Fini M,Mai A,Russo MA,Tafani M

    更新日期:2015-01-01 00:00:00

  • Dual inhibition of autophagy and the AKT pathway in prostate cancer.

    abstract::Genetic inactivation of PTEN through either gene deletion or mutation is common in metastatic prostate cancer, leading to activation of the phosphoinositide 3-kinase (PI3K-AKT) pathway, which is associated with poor clinical outcomes. The PI3K-AKT pathway plays a central role in various cellular processes supporting c...

    journal_title:Autophagy

    pub_type: 杂志文章

    doi:10.4161/auto.24921

    authors: Lamoureux F,Zoubeidi A

    更新日期:2013-07-01 00:00:00

  • MLKL contributes to Western diet-induced liver injury through inhibiting autophagy.

    abstract::Macroautophagy/autophagy is critical in maintaining cellular functions and homeostasis. Dynamic regulation of autophagy is associated with development of nonalcoholic fatty liver disease (NAFLD) and nonalcoholic steatohepatitis (NASH); however, the mechanisms involved in the regulation of autophagy in NAFLD/NASH are n...

    journal_title:Autophagy

    pub_type: 杂志文章

    doi:10.1080/15548627.2020.1760624

    authors: Wu X,Nagy LE

    更新日期:2020-07-01 00:00:00

  • Progesterone receptor membrane component 1/Sigma-2 receptor associates with MAP1LC3B and promotes autophagy.

    abstract::Autophagy resembles a recycling process in which proteins, organelles, or regions of the cytoplasm are enveloped and degraded. We have found that two of the central autophagy proteins, MAP1LC3 (microtubule-associated protein 1 light chain 3, also described as LC3) and UVRAG (UV radiation resistance associated/UV radia...

    journal_title:Autophagy

    pub_type: 杂志文章

    doi:10.4161/auto.25889

    authors: Mir SU,Schwarze SR,Jin L,Zhang J,Friend W,Miriyala S,St Clair D,Craven RJ

    更新日期:2013-10-01 00:00:00

  • Inhibition of autophagy as a therapeutic strategy of iron-induced brain injury after hemorrhage.

    abstract::Premenopausal women have better survival than men after intracerebral hemorrhage, which is associated with iron overproduction and autophagy induction. To examine the participation of neuronal autophagy and estrogen receptor α (ERα) in the E 2-mediated protection, PC12 neurons treated with Atg7 (autophagy-related prot...

    journal_title:Autophagy

    pub_type: 杂志文章

    doi:10.4161/auto.21289

    authors: Chen CW,Chen TY,Tsai KL,Lin CL,Yokoyama KK,Lee WS,Chiueh CC,Hsu C

    更新日期:2012-10-01 00:00:00

  • Promoting basal levels of autophagy in the nervous system enhances longevity and oxidant resistance in adult Drosophila.

    abstract::Autophagy is involved with the turnover of intracellular components and the management of stress responses. Genetic studies in mice have shown that suppression of neuronal autophagy can lead to the accumulation of protein aggregates and neurodegeneration. However, no study has shown that increasing autophagic gene exp...

    journal_title:Autophagy

    pub_type: 杂志文章

    doi:10.4161/auto.5269

    authors: Simonsen A,Cumming RC,Brech A,Isakson P,Schubert DR,Finley KD

    更新日期:2008-02-01 00:00:00

  • Perturbation of autophagic pathway by hepatitis C virus.

    abstract::Autophagy removes long-lived proteins and damaged organelles and is important for maintaining cellular homeostasis. It can also serve in innate immunity to remove intracellular pathogens. As such, viruses have evolved different mechanisms to subvert this innate immune response. We have recently demonstrated that hepat...

    journal_title:Autophagy

    pub_type: 杂志文章

    doi:10.4161/auto.6566

    authors: Sir D,Liang C,Chen WL,Jung JU,Ou JH

    更新日期:2008-08-01 00:00:00

  • Growth factor signaling permits hypoxia-induced autophagy by a HIF1alpha-dependent, BNIP3/3L-independent transcriptional program in human cancer cells.

    abstract::Several recent reports have demonstrated that autophagy is induced in response to hypoxia in cultured cells. However, the mechanism and consequence of hypoxia-induced autophagy remains unclear as there is no consensus between these studies. In our recent report we show that, in human cancer cells, hypoxia cooperates w...

    journal_title:Autophagy

    pub_type: 杂志文章

    doi:10.4161/auto.5.7.9821

    authors: Wilkinson S,Ryan KM

    更新日期:2009-10-01 00:00:00

  • Essential control of mitochondrial morphology and function by chaperone-mediated autophagy through degradation of PARK7.

    abstract::As a selective degradation system, chaperone-mediated autophagy (CMA) is essential for maintaining cellular homeostasis and survival under stress conditions. Increasing evidence points to an important role for the dysfunction of CMA in the pathogenesis of Parkinson disease (PD). However, the mechanisms by which CMA re...

    journal_title:Autophagy

    pub_type: 杂志文章

    doi:10.1080/15548627.2016.1179401

    authors: Wang B,Cai Z,Tao K,Zeng W,Lu F,Yang R,Feng D,Gao G,Yang Q

    更新日期:2016-08-02 00:00:00

  • Autophagy-inducing peptides from mammalian VSV and fish VHSV rhabdoviral G glycoproteins (G) as models for the development of new therapeutic molecules.

    abstract::It has not been elucidated whether or not autophagy is induced by rhabdoviral G glycoproteins (G) in vertebrate organisms for which rhabdovirus infection is lethal. Our work provides the first evidence that both mammalian (vesicular stomatitis virus, VSV) and fish (viral hemorrhagic septicemia virus, VHSV, and spring ...

    journal_title:Autophagy

    pub_type: 杂志文章

    doi:10.4161/auto.29557

    authors: García-Valtanen P,Ortega-Villaizán Mdel M,Martínez-López A,Medina-Gali R,Pérez L,Mackenzie S,Figueras A,Coll JM,Estepa A

    更新日期:2014-09-01 00:00:00

  • Autophagic degradation of an oncoprotein.

    abstract::Acute promyelocytic leukemia (APL) is characterized by a chromosomal t(15;17) translocation that fuses the gene encoding the promyelocytic leukemia protein (PML) to that encoding retinoic acid receptor alpha (RARA). The product of this genetic aberration, the PML/RARA fusion protein, is highly oncogenic and supports m...

    journal_title:Autophagy

    pub_type: 杂志文章

    doi:10.4161/auto.6.7.13066

    authors: Bøe SO,Simonsen A

    更新日期:2010-10-01 00:00:00