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Getting Rid of the Holidays Act I. Dairy Day. Word Metamorphism. The Nutty Professor. Voice Processing Module.

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By the use of the word "transcend", we imply a connection of play with the spiritual: play creates an artificial space beyond that of ordinary life. Play and ritual are braided together. Play brings order, ritual as well. Rituals build a hierarchy where the Gods are at a higher level and through expiation humans reach a balance with the Cosmos. What happens when play meets ritual?

A performance full of symbolism is born where the players represent the opposing forces of Nature fighting to reach a balance, stretching their limits, seeking for the stability of the world where they live. Tana Loch-Watts Tel. Ute Schoppet Tel. Vice-head of dept. Albert C. Joy Marlene Albers. Ralph Nickell Albert. Dean G Alberts.

Emilio Albrecht. Luiza Albrecht. Ann Miriam Albrich. Alanson Albright. Henry J. Clara M. Apsilah Daniel Albritton. George V Albu. Jane Albu. Wilhelm Albus. Uvergilio Alchini. Eronides Alchini Lorenzi. Robert Alcorn. Cathleen Alcorn. Maria Aguida V. Frank M Aldrich. Arthur Aldridge. Maria Grazia Alesso. Martha Kepner Townsend Alexander. Subby Alexander. Ida Alexander. William Alexander. J Alexander. Ella Alexander.

Frank Alfonsett. Josephine Alfonsett. William J Alford. Betty H Alford. Grace M Alford. H Algar. Leslie F. Ethel May Allan. James P. Suzanne C. Joseph Allano. Hugh Frederick Allcock. Edith B Allcorn. Mary Ann Allder. Michel Allemandi. Bernard V Allen. Arlene L Allen. John Henry Allen. Mattie L. Francies A Allen. Taswell H Allen. T W Allen. Ethan J. Mary Allen Dusatko. Nellie Margaret Allen.

Lydia E. Allen Hinckley. Roger David Allen. William E Allen. H P Allen. J W Allen. Sarah Ann Allen. J Allen. Janice Marlene Allen. Joseph Allen. Margaret Allen. Agnes B Allen. Eddie Lee Allen. Mickey J Allen.

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Alma E Amoroso. Vlastimil Amort. Franklin Amrstrong. Marry Amundsen. Anders Amund Amundsen. Isabel C. Otilia S. Fermino M. Mary Anatois. Sarah Anatois. Aaron Anbinder. Frank R Andelt. Bazil Anders. Mary Anders. Lorraine Anita Andersen. Dorothy Mae Andersen. Gudrun Helene Andersen. Arne Andersen. Elisabeth Andersen. Martinus 'Bud' Andersen. Ethel R. Joyce E. Vernon A. Odd Brurberg Andersen. Arlo K. Lorraine M Anderson. Alipio Luiz Anderson. Margaret Anderson. John William Anderson. J S Anderson.

Michele A. Clifton Andrew Anderson. Ellen Anderson. Axel L. Lillie E. Anderson Larson. Alfred Anderson. Mary Anderson Wrede. Glen A. Gordon E. Charlotte Anderson. Russell Anderson. Olive Anderson. Helen Anderson. Edwin H Anderson. Thomas D. Marvin G. Margaret I. Doris E. Edgar M.

Bernice Anderson. Gerda Anderson Ecklund. Vanda P Anderson. Lucinda N Anderson. Gus Anderson. Jeron Anderson. Wiliam Christie Anderson. Violet Anderson. Anna B. Helen V. F Anderson. James S. Lillian M. Delores Louise Anderson. Robert I. Ernest V. Leah M. Rogers Anderson. Luther Anderson. Mary M Anderson.

Frances Anderson Sparks Park. Harry Adolf Andersson. Per Edvin Andersson. Antonio Andrade Dos Santos. Denis Andre. Thor-Otto Andreassen. Phyllis A Andresen. Dwayne H Andresen. Robert D Andreu. Ann Andreu. James Andrew. Alice Mary Andrew. Florence Andrew. W A Andrews. Goliath Andrews. John W Andrews. James Robert Andrews. Margaret Mary Andrews. James W. Melba W Andrews. Bertha Andrews. Mary Agnes Gilbert Andrews.

Mary Jean Andrews. Gilbert L Andrews. John S. Lillian S. James Garfield Andrews. Melvin Andrews. Louis Andrieux. Eugenia Andrykowska Galuba. Jerzy Andrykowski. Marianna Andrzejczak. Stanislaw Andrzejczak. Kazimiera Andrzejewska. Teresa Andrzejewska. Alfon Andrzejewska. Anna Andrzejewska. Jadwiga Andrzejewska. Maria Andrzejewska. Michal Andrzejewski. Florian Andrzejewski. Arnildo Francisco Angeli. Maria Candida Angelino. Edward B Angell. Mary Angeloro. Umberto Angeloro. Jacob Angioletti. Angela M.

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Play demands order, absolute and supreme. While we play, we transcend the immediate needs of life. By the use of the word "transcend", we imply a connection of play with the spiritual: play creates an artificial space beyond that of ordinary life. Play and ritual are braided together. Play brings order, ritual as well. Rituals build a hierarchy where the Gods are at a higher level and through expiation humans reach a balance with the Cosmos.

What happens when play meets ritual? A performance full of symbolism is born where the players represent the opposing forces of Nature fighting to reach a balance, stretching their limits, seeking for the stability of the world where they live.

Tana Loch-Watts Tel. Harold Charlie. Jerry W. Maxine L. J Marie. Peter H. Margaret Z. Anna Winona. Euclides S. Herbert J. Thomas A. Jonas C. John F. Joseph C. Eva J. Bobbie E. Ronnie G. Agnes A. John B. Asa B. Jean Marie? Van Elsen. Jesse A? Celia A Da Silva. Urho Ensio Aaltonen. Fanny Aaronowsky. Nicholas B Abare. Lucia Abare. Aristides Abariotes. Anne Marie Abariotes. Saiah Abbington. William Allen Abbott. Percival Reynold Abbott. Rebecca Nahas Abbott.

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Toivo Aleksanteri Aikioniemi. Millie Ailers Siclair. George W. J Ainscough. G Ainsworth. F Aitchison. T Aitken. John Malcolm Aitken. Katarzyna Ajnbacher. Jozef Ajnbacher. James E. Royce Aaron Akromis. Rashid A Al-Rashid. Beverly B Al-Rashid. Luiz Carlos Albano. Osvaldo Jorge Albano. Strone Forest Albee.

Columbus C. Eliza H. Joseph Albee. Jemima Albee. Albert C. Joy Marlene Albers. Ralph Nickell Albert. Dean G Alberts. Emilio Albrecht. Luiza Albrecht. Ann Miriam Albrich. Alanson Albright. Henry J. Clara M. Apsilah Daniel Albritton.

George V Albu. Jane Albu. Wilhelm Albus. Uvergilio Alchini. Eronides Alchini Lorenzi. Robert Alcorn. Cathleen Alcorn. Maria Aguida V. Frank M Aldrich. Arthur Aldridge. Maria Grazia Alesso. Martha Kepner Townsend Alexander. Subby Alexander. Ida Alexander. William Alexander. J Alexander. Ella Alexander. Frank Alfonsett. Josephine Alfonsett. William J Alford.

Betty H Alford. Grace M Alford. H Algar. Leslie F. Ethel May Allan. James P. Suzanne C. Joseph Allano. Hugh Frederick Allcock. Edith B Allcorn. Mary Ann Allder. Michel Allemandi. Bernard V Allen. Arlene L Allen. John Henry Allen. Mattie L. Francies A Allen. Taswell H Allen. T W Allen. Ethan J. Mary Allen Dusatko. Nellie Margaret Allen. Lydia E. Allen Hinckley. Roger David Allen. William E Allen. H P Allen. J W Allen.

Sarah Ann Allen. J Allen. Janice Marlene Allen. Joseph Allen. Margaret Allen. Agnes B Allen. Eddie Lee Allen. Mickey J Allen. Frank C. Margaret Allen Pribil. William B Allen. Hamlet Allen. F Allen. Thelma Carey Allen. Keith Robert Allen. John R Alley. Shirley M Alley. Ilmo Allgayer. Geci Allgayer. Audrey L.

Earl H. A E Allis. Robert J Allison. Harriet Ruth Allison. Kathleen M. Nannie Allred. Thelma Mae Alm. Mary Elizabeth Jean Alm. Louis E. Raffaele Aloi. Angela of Mary Alstock. W Alston. Leonard F Alterberger.

Cecilia Alteruthemeier. Isabel Alteruthemeier. Maria Altieri. Vincenzo Altieri. Nelson Altmann. Mary Esdras Altstock. Edia Aluf. Reny N. Maria O. Alves Cardoso. Julia Alves Borges. Patricio Alves Braga. Amazilda Alves da Silva. Adelina Alves de Carvalho. Joaquim Alves de Carvalho. Miguel Alves de Oliveira. Adorino Alves de Ramos. Edi Alves Dos Santos. Virgolino Alves Vieira. Alecio Amaral. Maddalena Amatore.

Salvatore Amatore. Theresa Amatore. Ralph Amatore. Kamia Ambere. Joseph Warren Ambrose. Nellie Ambrose Butler. Mary Lucide Amell. Wiliam Henry Ames. Pamelia R. Jane Elizabeth Ames. Peggy Ames. E Ames. Malcolm McEwen Ames. Elizabeth Ames Atwater. Joshua Tyler Amick. Harriet E Amidon Gilbert. R F T Ammon. Michael Francis Patrick Amon. Mark Hugh Thomas Amon.

Jackie Amon. Alcidio Amorim. Fabricio Amorim Patricio. Alma E Amoroso. Vlastimil Amort. Franklin Amrstrong. Marry Amundsen. Anders Amund Amundsen. Isabel C. Otilia S. Fermino M. Mary Anatois. Sarah Anatois. Aaron Anbinder. Frank R Andelt. Bazil Anders. Mary Anders. Lorraine Anita Andersen. Dorothy Mae Andersen. Gudrun Helene Andersen. Arne Andersen. Elisabeth Andersen. Martinus 'Bud' Andersen. Ethel R.

Joyce E. Vernon A. Odd Brurberg Andersen. Arlo K. Lorraine M Anderson. Alipio Luiz Anderson. Margaret Anderson. John William Anderson. J S Anderson. Michele A. Clifton Andrew Anderson. Ellen Anderson. Axel L. Lillie E. Anderson Larson. Alfred Anderson. Mary Anderson Wrede. Glen A. Gordon E. Charlotte Anderson. Russell Anderson. Olive Anderson. Helen Anderson. Edwin H Anderson. Thomas D.

Marvin G. Margaret I. Doris E. Edgar M. Bernice Anderson. Gerda Anderson Ecklund. Vanda P Anderson. Lucinda N Anderson. Gus Anderson. Jeron Anderson. Wiliam Christie Anderson. Violet Anderson. Anna B. Helen V. F Anderson. James S. Lillian M. Delores Louise Anderson. Robert I. Ernest V. Leah M. Rogers Anderson. Luther Anderson. Mary M Anderson. Frances Anderson Sparks Park. Harry Adolf Andersson. Per Edvin Andersson.

Antonio Andrade Dos Santos. Denis Andre. Thor-Otto Andreassen. Phyllis A Andresen. Dwayne H Andresen. Robert D Andreu. Ann Andreu. James Andrew. Alice Mary Andrew. Florence Andrew. W A Andrews. Goliath Andrews. John W Andrews. James Robert Andrews. Margaret Mary Andrews. James W. Melba W Andrews. Bertha Andrews. Mary Agnes Gilbert Andrews. Mary Jean Andrews. Gilbert L Andrews. John S. Lillian S. James Garfield Andrews. Melvin Andrews.

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Urban Antkowiak. Despina Antokas. Janice Lee Antonicelli. Hereno Manoel Antunes. Suddetto Anunciata. Jaco Anverst. Marie Elizabeth Anzek Pitko. Jose Anzini. Diana Kay Apala. Dani Marie Apala. Eero Aleksei Api.

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By the use of the word "transcend", we imply a connection of play with the spiritual: play creates an artificial space beyond that of ordinary life. Play and ritual are braided together. Play brings order, ritual as well. Rituals build a hierarchy where the Gods are at a higher level and through expiation humans reach a balance with the Cosmos.

What happens when play meets ritual? A performance full of symbolism is born where the players represent the opposing forces of Nature fighting to reach a balance, stretching their limits, seeking for the stability of the world where they live. Tana Loch-Watts Tel. Ute Schoppet Tel. Vice-head of dept.

The methods of the psychotherapeutical process are instruments, which help the patient gain a deeper knowledge of himself, his limits, his suffering and his internal resources in order to face his suffering and to restructure.

Post-secondary education at the Universities of Strasbourg and Vienna. Graduated in developmental psychology MSc. Family therapist in training SGST. My services target children, adults and families. They cover counselling, therapy and coaching for a. My approach aims at activating and developing individual resources thereby allowing every person to sustainably deal with difficulties and challenges and restore their personal, social and professional well-being.

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Hier unsere Idee:. Ab 18 Uhr bitten wir Alle Kinder sich am Martinslied zu singen. Wir freuen uns auf ein kurzes Wiedersehen mit euch bei der Abholung eurer Bestellung. Bis dahin Nun ist es leider doch noch passiert. Unserer Feuerwehr wurde von der Verbandsgemeinde jegliche Hilfe bei Martinsveranstaltungen untersagt.

Es ist einfach eine Tatsache. Muss es in der Bus- wartehalle so aussehen, wie es dies manchmal tut? Ich sage mal, nein, muss es nicht. Er kann es nicht alleine schaffen. Ich sichere euch zu, dass die Gemeinde dies alles besorgt. CoBelVO v. Wir empfehlen deutlich zu machen, dass die Spielplatznutzung nur aufrecht erhalten werden kann, wenn die Verhaltensregeln eingehalten werden. Sonst bitte nacheinander und so, dass jedes Kind an die Reihe kommt.

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See more of Bettingen - Eifel on Facebook. Log In. Forgotten account? Not Now. Visitor Posts. Stefanie Roth. Ferdinand Friedrich. See more. Helfen trotz Corona. Continue reading. Although some advanced cancer patients can benefit from chemotherapy, residual tumors often recur soon after treatment. It is believed that a sub-population of tumor cells is not sensitive to treatment, and might be the cause of tumor relapse [ 2 , 3 ].

The characteristics of these cells, however, remain largely unknown. Usually, the mature miRNA is the guide strand for regulation of gene expression, while the passenger strand is believed to be degraded and inactivated [ 5 , 6 ]. The mature miRNAs regulate gene expression by targeting mRNAs post-transcriptionally A great deal of evidence has indicated that microRNAs play a crucial role in regulating tumor proliferation, apoptosis, angiogenesis and metastasis [ 7 - 19 ]. They also play important roles in development of glioblastoma [ 20 - 22 ].

Through the down-regulation of target proteins, microRNAs can function as oncomirs or tumor suppressors in a cell-specific context. MiR is one of the most extensively studied clusters. This cluster and its paralogs have been shown to be associated with many malignancies such as breast cancer, liver cancer, colon cancer, lung cancer and lymphoma [ 23 - 25 ].

The miR cluster has six components which share common characteristics in structure but differ in functions [ 26 , 27 ]. Since each member in this cluster can mediate multiple pathways and act diversely, there is a pressing need to explore the precise role of each component. There are documented evidences that dysfunctions of microRNA are associated with the development of glioblastoma [ 28 ]. Thus, miR has emerged as a critical regulator in mediating the cellular function of glioblastoma.

Emerging studies suggest that many stress signals are responsible for altered microRNA expression and functions [ 31 - 33 ]. Some microRNAs can modify gene expression by cross-talking with the tumor micro-environment, and their expression can be altered in turn by distinct stress conditions such as hypoxia, oxidative stimulation or radiation [ 34 - 42 ]. In response to stress, tumor cells often change gene expression to facilitate their survival [ 43 ].

Interestingly, such effects could be only achieved in stressed conditions such as serum deprivation or chemotherapeutic drug treatment, yet miR reduced tumor growth by targeting murine double minute 2 MDM2 under normal circumstances. Because the retarded proliferation rate often decreases chemo-sensitivity, miRtransfected cells develop resistance to chemotherapy.

Hereby we show miR has a dual function in glioblastoma: it suppresses tumor cell growth in normal conditions, and it also promotes tumor cell survival in unfavorable conditions. This highlights a potential mechanism in the response of tumor cells to stress and chemotherapy. The cells were also treated with the chemotherapeutic Temozolomide, followed by analysis of miRp levels. Nutrition deprivation increased miR levels. U87 and U cells were also treated with chemo-drug Temozolomide, followed by analysis of miR levels.

Temozolomide treatment increased miR levels. The miRoverexpression cells displayed higher ability of survival than the control cells. The distances between the wounding centre and the front of the migrating cells vertical axis were measured for statistical analysis. The invasive cells were stained blue and were counted in 6 randomly selected fields under a light microscope.

Expression of miR promoted cell invasion. To determine the role of miR in glioblastoma cells, U87 and U cells were stably transfected with miR expressing plasmid. Control cell lines were also established by using a plasmid without the miR precursor sequence. We then tested the roles of miR in regulating cell survival. The abilities of survival and metastasis to large distance are often complied with tumors that are resistant to chemotherapeutic drug treatment.

In order to evaluate metastasis potential, cell migration and invasiveness were measured by wound scratch assay and transwell test. Transwell test was also performed in different serum combination inside and outside of chambers. In addition, our findings suggested that miR conferred survival advantage to glioblastoma cells in an unfavorable condition and increased cell motility accordingly. Tumor expansions rely on sufficient supply of oxygen and other essential nutrients.

By inducing angiogenesis, tumor cells avoid being starved and escape chemotherapy. Emerging data suggest that angiogenesis of glioblastoma involves the interactions between endothelial cells and tumor cells [ 45 ]. Tube-like structure formation is an assay widely used to study angiogenesis in vitro. Lower, formation of the tube-like structures was quantified. Increased survival was seen in the miRtransfected cells.

We further found that miR regulated cell response under chemotherapy. Herein, the half maximal inhibitory concentration IC50 was calculated and applied to long-term chemotherapy. U87 cells were co-transfected with miR plasmid and one of the luciferase constructs. Mutations were generated on the seed regions red color , resulting in two mutant constructs Luc-Pten-1mut and Luc-Pten-2mut. The luciferase reporter vector Luc and the vector harboring a non-related region G3R were used as controls.

Lower, the cells were grown on 6-well tissue culture dishes. Cell survival was determined. Lower, U87 cells stably transfected with miR were transiently transfected with PTEN expression construct or the control vector and cultured for different days as indicated for survival assay. It is generally thought that TSCs play a major role in tumor re-vascularization and re-aggregation, eventually leading to tumor relapse.

Although the definition of TSCs is still controversial, CD, a cell surface glycoprotein, has been used extensively as a marker of glioblastoma stem-like cells GSC. To confirm that these spheres were alive, we continued to maintain the spheres in serum-free medium or serum-containing medium. Another prominent character of GSCs is that they can undergo self-renewal and differentiate.

We then examined the tumorigenesis of glioblastoma spheroid using the colony formation assay. Cells expressing miR formed significantly larger spheres than those transfected with the mock control left. The sphere cultures were continued to be maintained in serum-free medium, which induced extensive cell death in the control cells, but not in the miRtransfected cells. Addition of FBS into the cultures induced cell adhesion to the plate, displaying survivability of the spheres right.

Spheres formed in the secondary plates were divided by the numbers formed in the primary plates to evaluate the formation of spheres in the secondary plates. Sensitivities of the cells to the drug were tested. Cells transfected with miR displayed resistance to Docetaxel-induced cell death. Cells transfected with miR displayed resistance to all drugs. GSCs are thought to play an important role in drug resistance.

Therefore, we investigated the effects of chemotherapeutic agents on glioblastoma cells. We also treated the miR and vector-transfected U87 cells with Docetaxel, Carmustine and Temozolomide, followed by analysis of sensitivities of the cells to these drugs. Additionally, we examined the expression of CD using flow cytometry.

Moreover, when plated in serum, these floating neurospheres could differentiate to adherent cells again. The miRtransfected adhesive cells still expressed higher levels of CD than the control cells Fig S2.

CD expression was higher in the miRtransfected cells than the control cells in serum-containing medium 0. Lower, Typical photos are shown. Our findings indicated that miR inhibited glioblastoma cell proliferation, which is in agreement with other studies revealing similar results on breast cancer cells [ 47 ]. Taken together, these results suggest that miR suppresses glioblastoma cell growth under normal circumstances.

We then sought to identify the targets that mediated miR suppressing glioblastoma cell growth. Taking advantage of the online databases and computational algorithms, we screened a series of genes that could promote cell proliferation. MDM2 is an oncogene which is highly expressed in glioblastoma and it widely participates in tumorigenesis and progression.

It is thought to inhibit the activation of p53, but it can also regulate tumor cell proliferation independently [ 48 ]. U87 cells were co-transfected with miR plasmid and one of the constructs. MDM2 level was down-regulated in miRtransfected cells. Mutations were generated on the seed regions of each target sequence red color , resulting in four mutant constructs Luc-Mdmmut, Luc-Mdmmut, and Luc-Mdmmut.

Asterisks indicate significant differences. We then validated whether MDM2 played an essential role in modulating U87 cell activities. To corroborate this result, we performed rescue experiments by transfecting U87 cells with MDM2 expression construct. Cell proliferation was determined by counting the cells on day 1, 3, 5, and 7.

Given the fact that tumors often develop as a result of an aberrant response to a stress signal, it is important to determine the molecular biological mechanism involved. Therefore, bevacizumab, a monoclonal antibody against VEGF, has been approved to treat glioblastoma. It is believed to be able to starve tumors by blocking their blood supply. Nevertheless, highly penetrant tumor growth patterns in bevacizumab-treated patients have been repeatedly documented [ 49 ]. Here we identify that miRtransfected glioblastoma cells survived longer under starved stress, with the potential to develop the tube-like structures of endothelial cells and to enrich GSCs.

These may facilitate angiogenesis and increase the number of TSCs. This is consistent with our previous data that elucidated that slower growing cells are more resistant to chemotherapy-induced cell death [ 53 ]. Currently, chemotherapeutic agents that are commonly used in treating glioblastoma act by interfering with DNA replication, such as temozolimide and carmustine. It is conceivable that fast growing tumor cells are more easily suffered from cytotoxic agents compared with slower growing cells.

MiR therefore can induce chemo-resistance on glioblastoma cells by slowing down their proliferation. In addition, a reduced cell proliferation rate also benefits cells under starved conditions because a slower metabolic rate requires limited nutritional supply.

Very recently, similar finding were reported on other microRNAs miR and miRa which potentially modulate the oxidative stress response in ovarian carcinogenesis [ 54 ]. It was concluded that miR and miRa promoted tumor growth but sensitized tumors to chemotherapy, which was in agreement with our perspectives.

These data support the emerging model of microRNA: a buffering function. This refers to a microRNA's ability to target several pathways as both a positive and a negative regulator [ 55 ]. Documented examples have shown that the buffering function of microRNA is critical to maintain homeostasis in the systemic network [ 33 , 42 ]. The ability of miR to induce generation of glioblastoma stem-like cells is another interesting finding of our work. Although the role of microRNAs in the development of TSC has been studied extensively, there is still no general agreement on the definitions of TSCs in vitro [ 57 , 58 ].

It is known that TSCs can both undergo self-renewal and differentiate into a spectrum of mature cells. Moreover, recent discoveries indicate that they are widely involved in tumor progression, therapy resistance and distant metastasis. In glioblastoma, serum-free medium is a well-established method to enrich GSCs which can be detected by CD expression [ 59 , 60 ]. Serum contains essential nutrition factors for tumor cell growth.

During tumor progression to an advanced stage, it could be deprived of serum, under the stress of growth factor deficiency. In this study we reported that miR not only increased CD positive cells when cultured in SFM, but also increased capacities of self-renewal and colony formation ability. More importantly, GSCs are often thought to be responsible for drug resistance, which may be another potential mechanism accounting for chemo-resistance in tumor cells over-expressing miR At last, we found that miR increased tumor cell migration and invasiveness, which can also be found in neural stem cells [ 60 ].

Taken together, miR induced the generation of GSCs which display stem-like behaviors in multiple ways. In summary, our findings reveal a novel mechanism of stress response in glioblastoma cells. This adds new insights to our knowledge about microRNAs as mediators in tumor development. It has practical implications on clinical diagnosis and treatment.

In glioblastoma patients, miR could be used as a predictive marker of response to chemotherapy and anti-angiogenesis treatment. Although further studies are needed on the prognostic value of miR, our data suggests surgery and not drug treatment as a better option for patients who show over-expression of miR Green fluorescence was used to monitor transfected cells.

For each binding site, two pairs of primers were used to clone the fragments of 3'UTR and mutant controls. Peng at York University. RT-PCR was performed as described previously [ 66 ]. The primers specific for mature miR were purchased from Qiagen. Human U6 RNA was used as control.

The cells were harvested and cell numbers were counted in different time points. The cell number was counted every other day after Trypan Blue staining. Migration studies were performed by wound scratch tests and transwell invasion tests respectively. The monolayer of cells was scraped linearly with pipette tips, washed to remove cell debris and replenished with fresh media.

Microscope images of the scratch test were captured at the beginning and at different intervals later. As described previously, the tube-like structures were observed and recorded by microscopic examination after 24 hours [ 67 , 68 ].

Western blot was performed as previously described [ 69 ]. For cell cycle analysis, cells in the logarithmic phase were harvested and washed twice in PBS. Anti-CD antibody Abcam, dilution was added and stained on ice for 30 minutes.

Flow cytometry analysis was conducted after 30 minutes and CD ratio was detected by FL4. The data was analyzed using FlowJo 9. Colony formation was assessed by mixing cells with 0. After four weeks, colonies were stained by Coomassie blue Bio-Rad and photographed.