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MiR-96´s Effect on Metastasis in Prostate Cancer

Ammouri, Haneen (2014) MOBN19 20132
Degree Projects in Molecular Biology
Abstract
MicroRNAs (miRNAs) are an exciting new field which has been connected to different cancers and cellular imbalances. In previous studies, our group has been able to find a correlation between miR-96 and prostate cancer (PC). Here we studied miR-96’s effect on the cell-cell adhesion protein E-cadherin’s RNA and protein expression levels upon miR-96 inhibition. Furthermore, because the bone is one of PC’s main metastatic destinations, we studied miR-96’s enhanced potential in migration and colonization in the presence of bone derived factors. Using a 3-dimentional matrigel, we were able to observe a higher number of colony formations in miR-96 transfected 22RV1 cells in MG63 supernatant. The importance of nutrition is also obvious as a... (More)
MicroRNAs (miRNAs) are an exciting new field which has been connected to different cancers and cellular imbalances. In previous studies, our group has been able to find a correlation between miR-96 and prostate cancer (PC). Here we studied miR-96’s effect on the cell-cell adhesion protein E-cadherin’s RNA and protein expression levels upon miR-96 inhibition. Furthermore, because the bone is one of PC’s main metastatic destinations, we studied miR-96’s enhanced potential in migration and colonization in the presence of bone derived factors. Using a 3-dimentional matrigel, we were able to observe a higher number of colony formations in miR-96 transfected 22RV1 cells in MG63 supernatant. The importance of nutrition is also obvious as a noticeable increase in colonization was observed when the cells were first introduced to MG63 supernatant during transfection and then cultivated in normal medium in matrigel. In addition, miR-96 transfected DU145 and 22RV1 cells displayed higher random movement of migration towards the chemo-attractant MG63; with an endothelial layer MS1 as a barrier. What is interesting, is the possible connection between E-cadherin and colony formation because E-cadherin and colony formation both increase upon miR-96 transfection. This suggests that miR-96 could possibly have different targets and pathways depending on the surrounding environment promoting either solid tumor growth or migration. In conclusion, determination of miR-96’s mechanism and targets within different stages of adherence and environments of PC can lead to the development of better therapeutic tools. (Less)
Popular Abstract
Non-Coding RNA influence in prostate cancer

Prostate cancer (PC) is the second leading cause of cancer-associated deaths in western countries. Most men diagnosed with PC are around the age of 65 and there are different therapy approaches. One of the more successful therapies is called androgen deprivation therapy, which is based on the fact that PC cells depend on androgen, a steroids hormone, for prostate cell development. However, after 3-5 years of this therapy patients may develop castration resistant prostate cancer (CRPC), by which PC cells no longer depend on androgen for their growth and can carry on causing cancer regression regardless of this therapy. The cells start to evade cell death and move on to metastasize in the body,... (More)
Non-Coding RNA influence in prostate cancer

Prostate cancer (PC) is the second leading cause of cancer-associated deaths in western countries. Most men diagnosed with PC are around the age of 65 and there are different therapy approaches. One of the more successful therapies is called androgen deprivation therapy, which is based on the fact that PC cells depend on androgen, a steroids hormone, for prostate cell development. However, after 3-5 years of this therapy patients may develop castration resistant prostate cancer (CRPC), by which PC cells no longer depend on androgen for their growth and can carry on causing cancer regression regardless of this therapy. The cells start to evade cell death and move on to metastasize in the body, mostly in the bone.

One of the reasons for cancer progression, as in the bone, is imbalanced microRNAs (miRNAs) levels. MiRNAs are an exciting new field of study since they have been connected to different types of cancer and cellular functional imbalances. They are small non-coding RNAs which have the ability to bind to other messenger RNAs (mRNAs) and inhibit their translation, by blocking translation or degrading them. In previous studies, our group has been able to find a correlation between a specific miRNA, miR-96, and prostate cancer. We went on further to see how miR-96 can effect PC cells’ metastasis, solid tumor growth and effect on an adhesion protein E-cadherin, which plays an important role in cancer development.

Our group had previously looked into the binding targets of miR-96 and found E-cadherin to be among the top five. E-cadherin is a cell-cell adhesion glycoprotein associated with cell linkage within tissues. Here we demonstrated that E-cadherin RNA and protein expression levels decreased by the inhibition of mir-96 and may increase upon addition of miR-96 to those cells. MiR-96 is thus expected to regulate E-cadherin, yet further studies must be done to understand its mechanism of binding and downstream pathway effects.

Furthermore, because the bone is one of PC’s metastatic destinations, we studied miR-96’s enhanced potential in migration and colonization of cancer cells in MG63 supernatant. This is the media MG63, fibroblast from an osteosarcoma (bone cancer), grew in containing MG63 secretions which would simulate what PC cells would take up at the bone cancer site.
We first set out to study the likelihood of having more colonies formed by mir-96 introduced cells in MG63 supernatant. Using a 3-dementional matrigel, we were able to observe a higher number of colonies formed by miR-96 transfected (introduced) cells in MG63 supernatant. We also added to some of the samples in MG63 supernatant their original growing media, RPMI which contained more nutrients and which we found helps in increasing colony numbers. This sheds a light on the importance of angiogenesis studies, which is when the tumor gets nutrition by forming new blood vessels from pre-existing vessels.

PC cells transfected with miR-96 also had a higher tendency to move towards MG63 even with an endothelial barrier between the cells and MG63 supernatant.

What is interesting is that E-cadherin shouldn’t be in favour of migration, since it endorses cell-cell adhesion. Yet it increases upon miR-96 transfection when the possibility of migration isn’t there. Connecting these findings together would suggest that miR-96 could possibly have different targets and pathways in the cell depending on the surrounding environment. When the PC cell is inside the tumor, it can increase E-cadherin to help the tumor grow. But once certain cells lose E-cadherin, miR-96 will work on pushing the cells to move and migrate elsewhere. Upon encountering the bone, it will be more prone to initially stay there and grow a secondary tumor at that site.

Determination of miR-96’s mechanism and targets within different stages and environments and attachment abilities (prostate, blood and bone) of PC will lead to the development of better prognostic and diagnostic markers using mir-96 as well as its valuable utilization as a therapeutic tool in prostate cancer.

Supervisor: Yvonne Ceder
Master´s Degree Project in Molecular Genetics 45 credits, 2014
Department of Biology, Lund University (Less)
Please use this url to cite or link to this publication:
author
Ammouri, Haneen
supervisor
organization
course
MOBN19 20132
year
type
H2 - Master's Degree (Two Years)
subject
language
English
id
4678975
date added to LUP
2014-09-24 15:13:12
date last changed
2014-09-24 15:13:12
@misc{4678975,
  abstract     = {{MicroRNAs (miRNAs) are an exciting new field which has been connected to different cancers and cellular imbalances. In previous studies, our group has been able to find a correlation between miR-96 and prostate cancer (PC). Here we studied miR-96’s effect on the cell-cell adhesion protein E-cadherin’s RNA and protein expression levels upon miR-96 inhibition. Furthermore, because the bone is one of PC’s main metastatic destinations, we studied miR-96’s enhanced potential in migration and colonization in the presence of bone derived factors. Using a 3-dimentional matrigel, we were able to observe a higher number of colony formations in miR-96 transfected 22RV1 cells in MG63 supernatant. The importance of nutrition is also obvious as a noticeable increase in colonization was observed when the cells were first introduced to MG63 supernatant during transfection and then cultivated in normal medium in matrigel. In addition, miR-96 transfected DU145 and 22RV1 cells displayed higher random movement of migration towards the chemo-attractant MG63; with an endothelial layer MS1 as a barrier. What is interesting, is the possible connection between E-cadherin and colony formation because E-cadherin and colony formation both increase upon miR-96 transfection. This suggests that miR-96 could possibly have different targets and pathways depending on the surrounding environment promoting either solid tumor growth or migration. In conclusion, determination of miR-96’s mechanism and targets within different stages of adherence and environments of PC can lead to the development of better therapeutic tools.}},
  author       = {{Ammouri, Haneen}},
  language     = {{eng}},
  note         = {{Student Paper}},
  title        = {{MiR-96´s Effect on Metastasis in Prostate Cancer}},
  year         = {{2014}},
}