USSJ6C-60-M MSc Research Project: CLK Inhibitors on PC3 Cells

This MSc research project progression report details the investigation into the effects of novel CLK inhibitors on prostate cancer cell lines, specifically PC3 cells. The project encountered initial delays due to risk assessment issues and suboptimal cell counts. A planned comparison between PC3 and D...Read More

MSc – Research Project – USSJ6C-60-M
Progression report
Name:
Student ID:
Supervisor:
Project title:
A. If there was a delay in starting the project please indicate in the space below the
reasons.
The delay in my project was due to two reasons. This delay was unexpected by my
supervisor and I but we tackled the situation. The first issue which delayed us for
about 2 weeks was that the lab manager didn’t allow me to start my project as I
lacked an updated risk assessment to match my project. It took them 2 weeks to
deal with this issue. Another reason was that I started the experiment with lower
number of cells and thus it affected my results, which weren’t significant. The amount
of cells I used was lower than the cells specified in protocol.
B. Has there been a complete or partial change to the project plan? If so please
summarise what the new project is and clearly indicate why a change was
necessary.
I was intrigued to run this project by using two different cell lines, which were PC3
and DU145. I wanted to build my project around the differences between these two
lines and I wanted to know the effect of CLK inhibitors on them. The usage of two
different cell lines would’ve given the project a new dynamic and would’ve changed
the fundamentals of my research regarding the project. However, the constricted
time disabled me to perform my project around the difference between the cell lines,
PC3 and DU145 and I had to perform the project using the PC3 cell line.
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2018-2019

C. Has the project uncovered some unexpected results and have these findings
altered the project plan in any way?
When I started my project with the PC3 cell line, I was expecting a certain result but I
also came across unexpected results that really took me for surprise. These
unexpected findings and results changed the course of the project plan and
established a new aim for me to fulfil by completing my research on the PC3 cell line.
The unexpected data that I came across was that the kinase inhibitors are involved
in the activity of altering splicing mechanism and thus this indicated that they could
be used for treatment of some types of cancer, like prostate cancer.
D. In the space below please identify the objectives and/or distinct work packages of
your project. The rationale for their delivery should be clearly identified. Which ones
have you already met and are you on track to delivering the remaining areas of
study? [200 words]
The aim of my project was to investigate and provide a detailed report of the effect of
novel CLK inhibitors on prostate cancer lines. This aim of my project was further,
elaborately divided into the following objectives:
1. The first being the effect of the SR protein phosphorylation and alternative
splicing of key cancer associated genes by the CLK inhibitors.
2. The second aim of the project was to expand my research on the effect of
CLK inhibitors on cell proliferation and apoptosis.
3. The third aim of the project was to monitor the effects of CLK inhibitors on cell
invasion and cell migration.
All these aims helped in finding the true aim of the project which as to monitor the
overall effect of CLK inhibitors on the cells, which could also involve possibility of
treatment of cancer.
The achievements that I have been able to cover so far are:
1. I have been able to perform cell culture.
2. The second achievement is that I have found a link between concentration of
CLK inhibitors and number of cells. If the concentration of CLK inhibitors is
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more, then the number of cells would decrease. I found this result by treating
PC3 cell line with CLK inhibitors.
E. What aspects of the project, if any, have been most problematic? How have you
and your supervisor planned to resolve these issues?
During the completion of this project I have faced some problematic situations, which
I have been able to resolve with some help. The issues are:
1. Initially I had an issue with performing cell culture.
2. The second issue was that I found it fairly difficult to count the cells. Number
of cells were important as the protocol of the project specified certain number
of cells to be used.
3. I also had problem while calculating the number of cells in each well. To
resolve this issue, my supervisor transferred me to Simon, a PHD student,
who helped me with the issues.
F. Please indicate what aspects of the study have gone well.
Apart from the problems that I faced, which I have handled and resolved the issues,
there are also aspects in my projects that have gone well. As I have specified earlier,
my transfer to a PHD student, Simon, I have learned a lot. With the help of Simon, I
have learnt and am able to perform cell culture without any issue. I have also
managed to count the cells, which is an important aspect as incorrect number of
cells can alter the findings. I have also managed to do the required and appropriate
calculations that are required for cell treatment.
G. With regard to your research activity to date please identify the salient findings
and their significance. [200 words]
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I have performed the project on the PC3 cell line and I have come up with a uniform
result regarding the concentration of CLK inhibitors the number of cells (PC3 cells). I
have come to conclusion that there is a link between the concentration of CLK
inhibitors and the number of PC3 cells. This indicates interdependency between
these two factors. My findings are in the order that the increase in the concentration
of CLK inhibitors, SU1411 has brought about or initiated an increase in the number
of cells and their viability. The reasons of my findings can be based on the
foundation that the CLK inhibitors that I have used are a type of protein. And this
proteinaceous substance, CLK inhibitor has suppressed the activity of the mRNA of
the PC3 cells. Due to the suppression of the mRNA of the PC3 cell line, the viability
and the activity of the cells have also decreased. The findings of this project
regarding the concentration of CLK concentration and its effect on the numbers and
the viability of cells can also be linked to other findings and results which could be
helpful in lining other experiments and projects.
H. Are there any new approaches or ideas that you would like to explore? Have you
discussed these new opportunities with your supervisor? If so how were they met?
I have really been intrigued by the findings of this project and I have been able to
achieve the results with apt help. The new approaches and ideas that I would like to
explore and research with regard to this project is to study and compare the
differences between TG003 and SU1411. I would also like to perform experiment on
different cell lines, which would bring out different results. I have discussed the
numerous possibilities and the intent to perform these experiments with my
supervisor and they too have been enthusiastic and intrigued on the prospects of this
new project.
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