INTERACTION BETWEEN BRK AND HER2 IN BREAST CANCER

INTERACTION BETWEEN BRK AND HER2 IN BREAST CANCER Midan Ai, Ph.D. Supervisory Professor: Zhen Fan, M.D. Breast tumor kinase (Brk) is a nonreceptor protein-tyrosine kinase that is highly expressed in approximately two thirds of breast cancers but is not detectable or is expressed at very low levels in normal mammary epithelium. Brk plays important roles in promoting proliferation, survival, invasion, and metastasis of breast cancer cells, but the mechanism(s) of which remain largely unknown. Recent studies showed that Brk is frequently co-overexpressed with human epidermal growth factor receptor-2 (HER2) and is physically associated with HER2 in breast cancer. The mechanism needs to be determined. In my studies, I found that high expression of HER2 is correlated with high expression of Brk in breast cancer cell lines. Silencing HER2 expression via RNA interference in HER2 over-expressed breast cancer cells resulted in Brk protein decrease and overexpression of HER2 in HER2 low-expressed breast cancer cells up-regulated Brk expression. The mechanism study indicated that overexpression of HER2 increased Brk protein stability. Brk was degraded through a Ca2+-dependent protease pathway involving calpain and HER2 stimulated Brk expression via inhibiting calpain activity. Calpastatin is a calpain endogenous inhibitor and the calpain-calpastatin system has been implicated in a number of cell physiological functions. HER2 restrained calpain activation via up-regulating calpastatin expression and HER2 downstream signaling, MAPK pathway, was involved in the regulation. Furthermore, silencing of Brk expression by RNA interference in HER2-overexpressing breast cancer cells decreased HER2-mediated cell proliferation, survival, invasion/metastasis potential and increased cell sensitivity to HER2 kinase inhibitor, lapatinib, treatment, indicating that Brk plays important roles in regulating and mediating the oncogenic functions of HER2. The Stat3 pathway played important roles in Brk mediated cell survival and invasion/metastasis in the context of HER2-overexpressing breast cancer cells. However, transgenic mice with inducible expression of constitutively active Brk (CA) in the mammary epithelium failed to develop malignant change in the mammary glands after Brk induction for 15 months which indicated that expression of Brk protein alone was not sufficiently to induce spontaneous breast tumor. Bitransgenic mice with co-expression of HER2/neu and inducible expression of Brk in the mammary epithelium developed multifocal mammary tumors, but there were no significant difference in the tumor occurring time, tumor size, tumor weight and tumor multiplicity between the mouse group with co-expression of Brk and HER2/neu and the mouse group with HER2/neu expression only.

Räkna med bråk : En litteraturstudie om viktiga aspekter för att elever i årskurs 4-6 skall utveckla förståelse för bråk

Syftet med denna systematiska litteraturstudie är att få kunskap om viktiga aspekter för att elever i årskurs 4-6 skall utveckla förståelse för bråk. Utifrån syftet formulerades följande frågeställning: Vilka undervisningsmetoder kan underlätta elevers förståelse av bråk. Databaserna LIBRIS, Teacher´s reference center och ERIC användes för söka efter relevanta vetenskapligt granskade artiklar. Resultatet visar att många elever tycker att det är svårt med bråk och att de inte förstår syftet med att lära sig bråk i skolan. Vidare visar resultatet att om lärarna åskådliggör bråk för eleverna på många olika sätt i undervisningen ökar elevernas förståelse för bråk. Studien visar även att användning av konkreta material såsom modeller och bilder gör det enklare för elever att bland annat förstå beräkningar med bråk. Att lärarna samtalar med eleverna om bråk är också en viktig aspekt för att öka elevers förståelse för bråk.

Räkna med bråk : En litteraturstudie om viktiga aspekter för att elever i årskurs 4-6 skall utveckla förståelse för bråk

Syftet med denna systematiska litteraturstudie är att få kunskap om viktiga aspekter för att elever i årskurs 4-6 skall utveckla förståelse för bråk. Utifrån syftet formulerades följande frågeställning: Vilka undervisningsmetoder kan underlätta elevers förståelse av bråk. Databaserna LIBRIS, Teacher´s reference center och ERIC användes för söka efter relevanta vetenskapligt granskade artiklar. Resultatet visar att många elever tycker att det är svårt med bråk och att de inte förstår syftet med att lära sig bråk i skolan. Vidare visar resultatet att om lärarna åskådliggör bråk för eleverna på många olika sätt i undervisningen ökar elevernas förståelse för bråk. Studien visar även att användning av konkreta material såsom modeller och bilder gör det enklare för elever att bland annat förstå beräkningar med bråk. Att lärarna samtalar med eleverna om bråk är också en viktig aspekt för att öka elevers förståelse för bråk.

Microstructural Evolution of A356 Al Alloy During Flow Along a Cooling Slope

Cooling slope (CS) has been used in this study to prepare semi-solid slurry of A356 Al alloy, keeping in view of slurry generation on demand for Rheo-pressure die casting process. Understanding the physics of microstructure evolution during cooling slope slurry formation is important to satisfy the need of semi-sold slurry with desired shape, size and morphology of primary Al phase. Mixture of spherical and rosette shaped primary Al phase has been observed in the samples collected during melt flow through the slope as well as in the cast (mould) samples compared to that of dendritic shape, observed in case of conventionally cast A356 alloy. The liquid melt has been poured into the slope at 650 A degrees C temperature and during flow it falls below the liquidus temperature of the said alloy, which facilitates crystallization of alpha-Al crystals on the cooling slope wall. Crystal separation due to melt flow is found responsible for nearly spherical morphology of the primary Al phase.

Spin-glass state in nanoparticulate (La0.7Sr0.3MnO3)(1-x) (BaTiO3)(x) solid solutions: Experimental and density-functional studies

We report the transition from robust ferromagnetism to a spin- glass state in nanoparticulate La0.7Sr0.3MnO3 through solid solution with BaTiO3. The field- and temperature-dependent magnetization and the frequency-dependent ac magnetic susceptibility measurements strongly indicate the existence of a spin- glass state in the system, which is further confirmed from memory effect measurements. The breaking of long-range ordering into short-range magnetic domains is further investigated using density-functional calculations. We show that Ti ions remain magnetically inactive due to insufficient electron leakage from La0.7Sr0.3MnO3 to the otherwise unoccupied Ti-d states. This results in the absence of a Mn-Ti-Mn spin exchange interaction and hence the breaking of the long-range ordering. Total-energy calculations suggest that the segregation of nonmagnetic Ti ions leads to the formation of short-range ferromagnetic Mn domains.

Identification of target genes of the T-box proteins in Drosophila melanogaster

In a prior bioinformatic analysis by Hüyseyin Binbas, potential Tbx targets sequences in wing-related genes have been identified. Guided by this information, enhancer trap/reporter lacZ insertions were characterized by X-gal staining first in wildtype and then in l(1)omb imaginal discs.rnIn several lines I observed an increase in reporter expression in a l(1)omb mutant background. Since Omb is assumed to function predominantly as a transcriptional repressor, this may indicate direct regulation. Repression by Omb was observed e.g. for brk and tkv. These genes are negatively regulated by Dpp, while omb is induced by Dpp. Omb which mediates the effects of Dpp on proliferation could, thus, also mediate the Dpp effect on patterning of the wing disc. However, brk and tkv were not completely derepressed in l(1)omb indicating that Dpp represses these genes also by an Omb-independent mechanism.rnMore frequently I observed loss of reporter expression in an l(1)omb mutant background. In these cases, regulation by Omb presumably is indirect. For example, STAT92E-lacZ expression in the wildtype eye was symmetrically expressed at the dorsal and ventral margins. In l(1)omb, ventral expression was selectively lost. Loss of omb is known to cause ventral overproliferation of the eye by activation of the Jak/STAT pathway. STAT92E expression is negatively regulated by Jak/STAT signaling suggesting that loss of omb activates Jak/STAT further upstream in the pathway.rnRegional overproliferation of eye and wing in the l(1)omb mutant background proved a complicating issue in the search for Omb targets. This effect made it difficult to decide whether an expanded reporter expression pattern was due to tissue expansion or reporter gene derepression. For instance hth-lacZ appeared to expand along the ventral eye disc margin in l(1)omb. Without addtional experiments it cannot be concluded whether this is due to de-repression or to activation in association with the proliferative state. Parallel to my experiments, evidence accumulated in our laboratory that loss of omb may attenuate Wg and Hegehog signaling. Since these diffusible proteins are the main patterning molecules in the wing imaginal disc, with dpp being downstream of Hh, many of the observed effects could be secondary to reduced Wg and Hh activity. Examples are ab-lacZ, Dll-lacZ and vgBE-lacZ (reduced expression on the dorso-ventral boundary) and inv-lacZ (late larval expression in the anterior wing disc compartment is lost) or sal-lacZ. Epistasis experiment will be required to clarifiy these issues.rnFurthermore, loss of omb appeared to induce cell fate changes. It was reported previously that in an omb null mutant, the dorsal determinant apterous (ap) is ectopically expressed in the ventral compartment (an effect I did not observe with the strongly hypomorphic l(1)omb15, indicating strong dose dependence). Ventral repression of ap is maintained by epigenetic mechanisms. The patchy and variable nature of ectopic expression of ap or grn-1.1-lacZ points to an effect of omb on epigenetic stability.rnIn the second part of my thesis, an analysis of Omb expression in the Drosophila embryonic ventral nervous system was performed. Omb was found co-expressed with Eve in the medial aCC and RP2 motorneurons as well as the fpCC interneuron and the mediolateral CQ neurons. Additionally, Omb was detected in the Eg positive NB7-3 GW serotonergic motoneuron and the N2-4 neurons. Omb was not found in Repo positive glial cells. During embryonic stage 14, Omb showed some coepression with Dpn or Pros. At the embryonic stage 16, Omb was expressed in minor subset of Mid and Wg positive cells.